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Sample records for reliable simultaneous detection

  1. Simultaneous amplification of two bacterial genes: more reliable method of Helicobacter pylori detection in microbial rich dental plaque samples.

    Science.gov (United States)

    Chaudhry, Saima; Idrees, Muhammad; Izhar, Mateen; Butt, Arshad Kamal; Khan, Ayyaz Ali

    2011-01-01

    Polymerase Chain reaction (PCR) assay is considered superior to other methods for detection of Helicobacter pylori (H. pylori) in oral cavity; however, it also has limitations when sample under study is microbial rich dental plaque. The type of gene targeted and number of primers used for bacterial detection in dental plaque samples can have a significant effect on the results obtained as there are a number of closely related bacterial species residing in plaque biofilm. Also due to high recombination rate of H. pylori some of the genes might be down regulated or absent. The present study was conducted to determine the frequency of H. pylori colonization of dental plaque by simultaneously amplifying two genes of the bacterium. One hundred dental plaque specimens were collected from dyspeptic patients before their upper gastrointestinal endoscopy and presence of H. pylori was determined through PCR assay using primers targeting two different genes of the bacterium. Eighty-nine of the 100 samples were included in final analysis. With simultaneous amplification of two bacterial genes 51.6% of the dental plaque samples were positive for H. pylori while this prevalence increased to 73% when only one gene amplification was used for bacterial identification. Detection of H. pylori in dental plaque samples is more reliable when two genes of the bacterium are simultaneously amplified as compared to one gene amplification only.

  2. Robust simultaneous detection of coronary borders in complex images

    International Nuclear Information System (INIS)

    Sonka, M.; Winniford, M.D.; Collins, S.M.

    1995-01-01

    Visual estimation of coronary obstruction severity from angiograms suffers from poor inter- and intraobserver reproducibility and is often inaccurate. In spite of the widely recognized limitations of visual analysis, automated methods have not found widespread clinical use, in part because they too frequently fail to accurately identify vessel borders. The authors have developed a robust method for simultaneous detection of left and right coronary borders that is suitable for analysis of complex images with poor contrast, nearby or overlapping structures, or branching vessels. The reliability of the simultaneous border detection method and that of their previously reported conventional border detection method were tested in 130 complex images, selected because conventional automated border detection might be expected to fail. Conventional analysis failed to yield acceptable borders in 65/130 or 50% of images. Simultaneous border detection was much more robust (p < .001) and failed in only 15/130 or 12% of complex images. Simultaneous border detection identified stenosis diameters that correlated significantly better with observer-derived stenosis diameters than did diameters obtained with conventional border detection (p < 0.001). Simultaneous detection of left and right coronary borders is highly robust and has substantial promise for enhancing the utility of quantitative coronary angiography in the clinical setting

  3. Simultaneous detection of indicators of hepatitis virus exposure

    International Nuclear Information System (INIS)

    Ling, C.; Decker, R.A.; Overby, L.R.

    1981-01-01

    This invention discloses an improvement in solid phase immunoassay methods for the detection and determination of antigens and antibodies (markers) relating to hepatitis. The method for simultaneously detecting in a sample at least two different markers evidencing exposure to hepatitis virus comprises contacting the sample with a solid phase reagent which is coated with at least two different, non-complementary immunoreactants which are complementary to the unknown markers to be detected, then with a liquid reagent comprising at least two different hepatitis markers or immunoreactants, each selected to either react or compete with one of the unknown markers and each labeled with a detectably distinct tag. Examples described use 125 I. (author)

  4. Fast simultaneous electrochemical detection of tetracycline and fluoxetine in water

    NARCIS (Netherlands)

    Ardelean, Magdalena; Pode, Rodica; Schoonman, J.; Pop, Aniela; Manea, Florica

    2017-01-01

    The electrochemical methods-based protocol for simultaneous detection of tetracycline (TC) from antibiotics class and fluoxetine (FXT) from anti-depressive pharmaceuticals class, which belongs to emerging pollutants from water, was developed in this study using carbon nanofiber-epoxy composite

  5. Reliability of leak detection systems in LWRs

    International Nuclear Information System (INIS)

    Kupperman, D.S.

    1986-10-01

    In this paper, NRC guidelines for leak detection will be reviewed, current practices described, potential safety-related problems discussed, and potential improvements in leak detection technology (with emphasis on acoustic methods) evaluated

  6. New Multiplexing Tools for Reliable GMO Detection

    NARCIS (Netherlands)

    Pla, M.; Nadal, A.; Baeten, V.; Bahrdt, C.; Berben, G.; Bertheau, Y.; Coll, A.; Dijk, van J.P.; Dobnik, D.; Fernandez-Pierna, J.A.; Gruden, K.; Hamels, S.; Holck, A.; Holst-Jensen, A.; Janssen, E.; Kok, E.J.; Paz, La J.L.; Laval, V.; Leimanis, S.; Malcevschi, A.; Marmiroli, N.; Morisset, D.; Prins, T.W.; Remacle, J.; Ujhelyi, G.; Wulff, D.

    2012-01-01

    Among the available methods for GMO detection, enforcement and routine laboratories use in practice PCR, based on the detection of transgenic DNA. The cost required for GMO analysis is constantly increasing due to the progress of GMO commercialization, with inclusion of higher diversity of species,

  7. Reliably detectable flaw size for NDE methods that use calibration

    Science.gov (United States)

    Koshti, Ajay M.

    2017-04-01

    Probability of detection (POD) analysis is used in assessing reliably detectable flaw size in nondestructive evaluation (NDE). MIL-HDBK-1823 and associated mh18232 POD software gives most common methods of POD analysis. In this paper, POD analysis is applied to an NDE method, such as eddy current testing, where calibration is used. NDE calibration standards have known size artificial flaws such as electro-discharge machined (EDM) notches and flat bottom hole (FBH) reflectors which are used to set instrument sensitivity for detection of real flaws. Real flaws such as cracks and crack-like flaws are desired to be detected using these NDE methods. A reliably detectable crack size is required for safe life analysis of fracture critical parts. Therefore, it is important to correlate signal responses from real flaws with signal responses form artificial flaws used in calibration process to determine reliably detectable flaw size.

  8. Multiplex PCR assay for simultaneous detection of six major bacterial pathogens of rice.

    Science.gov (United States)

    Cui, Z; Ojaghian, M R; Tao, Z; Kakar, K U; Zeng, J; Zhao, W; Duan, Y; Vera Cruz, C M; Li, B; Zhu, B; Xie, G

    2016-05-01

    The aim of this study was to develop a multiplex PCR (mPCR) assay for rapid, sensitive and simultaneous detection of six important rice pathogens: Xanthomonas oryzae pv. oryzae, X. oryzae pv. oryzicola, Pseudomonas fuscovaginae, Burkholderia glumae, Burkholderia gladioli and Acidovorax avenae subsp. avenae. Specific primers were designed through a bioinformatics pipeline. Sensitivity of detection was established using both traditional PCR and quantitative real-time PCR on isolated DNA and on bacterial cells both in vitro and in simulated diseased seeds and the parameters were optimized for an mPCR assay. A total of 150 bacterial strains were tested for specificity. The mPCR assay accurately predicted the presence of pathogens among 44 symptomatic and asymptomatic rice seed, sheath and leaf samples. This study confirmed that this mPCR assay is a rapid, reliable and simple tool for the simultaneous detection of six important rice bacterial pathogens. This study is the first report of a method allowing simultaneous detection of six major rice pathogens. The ability to use crude extracts from plants without bacterial isolation or DNA extraction enhances the value of this mPCR technology for rapid detection and aetiological/epidemiological studies. © 2016 The Society for Applied Microbiology.

  9. Simultaneous detection of three lily viruses using Triplex IC-RT-PCR.

    Science.gov (United States)

    Zhang, Yubao; Wang, Yajun; Xie, Zhongkui; Yang, Guo; Guo, Zhihong; Wang, Le

    2017-11-01

    Viruses commonly infecting lily (Lilium spp.) include: Lily symptomless virus (LSV), Cucumber mosaic virus (CMV) and Lily mottle virus (LMoV). These viruses usually co-infect lilies causing severe economic losses in terms of quantity and quality of flower and bulb production around the world. Reliable and precise detection systems need to be developed for virus identification. We describe the development of a triplex immunocapture (IC) reverse transcription (RT) polymerase chain reaction (PCR) assay for the simultaneous detection of LSV, CMV and LMoV. The triplex IC-RT-PCR was compared with a quadruplex RT-PCR assay. Relative to the quadruplex RT-PCR, the specificity of the triplex IC-RT-PCR system for LSV, CMV and LMoV was 100% for field samples. The sensitivity of the triplex IC-RT-PCR system was 99.4%, 81.4% and 98.7% for LSV, CMV and LMoV, respectively. Agreement (κ) between the results obtained from the two tests was 0.968, 0.844 and 0.984 for LSV, CMV and LMoV, respectively. This is the first report of the simultaneous detection of LSV, CMV and LMoV in a triplex IC-RT-PCR assay. In particular we believe this convenient and reliable triplex IC-RT-PCR method could be used routinely for large-scale field surveys or crop health monitoring of lily. Copyright © 2017. Published by Elsevier B.V.

  10. Silver surface enrichment controlled by simultaneous RBS for reliable PIXE analysis of ancient coins

    International Nuclear Information System (INIS)

    Beck, L.; Alloin, E.; Berthier, C.; Reveillon, S.; Costa, V.

    2008-01-01

    Evidence of silver surface enrichment of ancient silver-copper coins has been pointed out in the past years. Surface enrichment can be fortuitous or intentional. In this paper, we have investigated the cleaning procedures usually performed after excavation or in museums. We have shown that chemicals or commercial products routinely used dissolve preferentially the copper phase and consequently contribute to the silver surface enrichment. As a result, surface analyses such as PIXE or XRF can be strongly affected by this effect. By using simultaneously RBS and PIXE, it is possible to check through the silver surface enrichment and then select the reliable measurements, characteristic of the bulk composition. Results on coins recently discovered and mechanically or chemically cleaned are presented

  11. UTILIZATION OF PHOSWICH DETECTORS FOR SIMULTANEOUS, MULTIPLE RADIATION DETECTION

    International Nuclear Information System (INIS)

    Miller, William H.; Manuel Diaz de Leon

    2003-01-01

    A phoswich radiation detector is comprised of a phosphor sandwich in which several different phosphors are viewed by a common photomultiplier. By selecting the appropriate phosphors, this system can be used to simultaneously measure multiple radiation types (alpha, beta, gamma and/or neutron) with a single detector. Differentiation between the signals from the different phosphors is accomplished using digital pulse shape discrimination techniques. This method has been shown to result in accurate discrimination with highly reliable and versatile digital systems. This system also requires minimal component count (i.e. only the detector and a computer for signal processing). A variety of detectors of this type have been built and tested including: (1) a triple phoswich system for alpha/beta/gamma swipe counting, (2) two well-type detectors for measuring low levels of low energy photons in the presence of a high energy background, (3) a large area detector for measuring beta contamination in the presence of a photon background, (4) another large area detector for measuring low energy photons from radioactive elements such as uranium in the presence of a photon background. An annular geometry, triple phoswich system optimized for measuring alpha/beta/gamma radiation in liquid waste processing streams is currently being designed

  12. UTILIZATION OF PHOSWICH DETECTORS FOR SIMULTANEOUS, MULTIPLE RADIATION DETECTION

    Energy Technology Data Exchange (ETDEWEB)

    William H. Miller; Manuel Diaz de Leon

    2003-04-15

    A phoswich radiation detector is comprised of a phosphor sandwich in which several different phosphors are viewed by a common photomultiplier. By selecting the appropriate phosphors, this system can be used to simultaneously measure multiple radiation types (alpha, beta, gamma and/or neutron) with a single detector. Differentiation between the signals from the different phosphors is accomplished using digital pulse shape discrimination techniques. This method has been shown to result in accurate discrimination with highly reliable and versatile digital systems. This system also requires minimal component count (i.e. only the detector and a computer for signal processing). A variety of detectors of this type have been built and tested including: (1) a triple phoswich system for alpha/beta/gamma swipe counting, (2) two well-type detectors for measuring low levels of low energy photons in the presence of a high energy background, (3) a large area detector for measuring beta contamination in the presence of a photon background, (4) another large area detector for measuring low energy photons from radioactive elements such as uranium in the presence of a photon background. An annular geometry, triple phoswich system optimized for measuring alpha/beta/gamma radiation in liquid waste processing streams is currently being designed.

  13. Rapid and reliable detection and identification of GM events using multiplex PCR coupled with oligonucleotide microarray.

    Science.gov (United States)

    Xu, Xiaodan; Li, Yingcong; Zhao, Heng; Wen, Si-yuan; Wang, Sheng-qi; Huang, Jian; Huang, Kun-lun; Luo, Yun-bo

    2005-05-18

    To devise a rapid and reliable method for the detection and identification of genetically modified (GM) events, we developed a multiplex polymerase chain reaction (PCR) coupled with a DNA microarray system simultaneously aiming at many targets in a single reaction. The system included probes for screening gene, species reference gene, specific gene, construct-specific gene, event-specific gene, and internal and negative control genes. 18S rRNA was combined with species reference genes as internal controls to assess the efficiency of all reactions and to eliminate false negatives. Two sets of the multiplex PCR system were used to amplify four and five targets, respectively. Eight different structure genes could be detected and identified simultaneously for Roundup Ready soybean in a single microarray. The microarray specificity was validated by its ability to discriminate two GM maizes Bt176 and Bt11. The advantages of this method are its high specificity and greatly reduced false-positives and -negatives. The multiplex PCR coupled with microarray technology presented here is a rapid and reliable tool for the simultaneous detection of GM organism ingredients.

  14. Reliability evaluation of the Savannah River reactor leak detection system

    International Nuclear Information System (INIS)

    Daugherty, W.L.; Sindelar, R.L.; Wallace, I.T.

    1991-01-01

    The Savannah River Reactors have been in operation since the mid-1950's. The primary degradation mode for the primary coolant loop piping is intergranular stress corrosion cracking. The leak-before-break (LBB) capability of the primary system piping has been demonstrated as part of an overall structural integrity evaluation. One element of the LBB analyses is a reliability evaluation of the leak detection system. The most sensitive element of the leak detection system is the airborne tritium monitors. The presence of small amounts of tritium in the heavy water coolant provide the basis for a very sensitive system of leak detection. The reliability of the tritium monitors to properly identify a crack leaking at a rate of either 50 or 300 lb/day (0.004 or 0.023 gpm, respectively) has been characterized. These leak rates correspond to action points for which specific operator actions are required. High reliability has been demonstrated using standard fault tree techniques. The probability of not detecting a leak within an assumed mission time of 24 hours is estimated to be approximately 5 x 10 -5 per demand. This result is obtained for both leak rates considered. The methodology and assumptions used to obtain this result are described in this paper. 3 refs., 1 fig., 1 tab

  15. Simultaneous Detection of Barley Virus Diseases in Korea

    Directory of Open Access Journals (Sweden)

    Bong-Choon Lee

    2017-12-01

    Full Text Available Barley mild mosaic virus (BaMMV, Barley yellow mosaic virus (BaYMV and Barley yellow dwarf virus (BYDV have been identified as an important causative agents for an economically important disease of winter barley in Korea. In this study, a multiplex reverse transcription polymerase chain reaction (mRT-PCR method was used for the simultaneous detection. Three sets of virus-specific primers targeted to the capsid protein coding genes of BaMMV, BaYMV and BYDV were used to amplify fragments that were 594 bp, 461 bp, and 290 bp, respectively. Several sets of primers for each target virus were evaluated for their sensitivity and specificity by multiplex RT-PCR. The optimum primer concentrations and RT-PCR conditions were determined for the multiplex RT-PCR. The mRT-PCR assay was found to be a better and rapid virus diagnostic tool of specific barley diseases and potential for investigating the epidemiology of these viral diseases.

  16. Simultaneous detection of imidacloprid and parathion by the dual-labeled time-resolved fluoroimmunoassay.

    Science.gov (United States)

    Shi, Haiyan; Sheng, Enze; Feng, Lu; Zhou, Liangliang; Hua, Xiude; Wang, Minghua

    2015-10-01

    A highly sensitive direct dual-labeled time-resolved fluoroimmunoassay (TRFIA) to detect parathion and imidacloprid simultaneously in food and environmental matrices was developed. Europium (Eu(3+)) and samarium (Sm(3+)) were used as fluorescent labels by coupling separately with L1-Ab and A1P1-Ab. Under optimal assay conditions, the half-maximal inhibition concentration (IC50) and limit of detection (LOD, IC10) were 10.87 and 0.025 μg/L for parathion and 7.08 and 0.028 μg/L for imidacloprid, respectively. The cross-reactivities (CR) were negligible except for methyl-parathion (42.4 %) and imidaclothiz (103.4 %). The average recoveries of imidacloprid ranged from 78.9 to 104.2 % in water, soil, rice, tomato, and Chinese cabbage with a relative standard deviation (RSD) of 2.4 to 11.6 %, and those of parathion were from 81.5 to 110.9 % with the RSD of 3.2 to 10.5 %. The results of TRFIA for the authentic samples were validated by comparison with gas chromatography (GC) analyses, and satisfactory correlations (parathion: R (2) = 0.9918; imidacloprid: R (2) = 0.9908) were obtained. The results indicate that the dual-labeled TRFIA is convenient and reliable to detect parathion and imidacloprid simultaneously in food and environmental matrices.

  17. Fault detection and reliability, knowledge based and other approaches

    International Nuclear Information System (INIS)

    Singh, M.G.; Hindi, K.S.; Tzafestas, S.G.

    1987-01-01

    These proceedings are split up into four major parts in order to reflect the most significant aspects of reliability and fault detection as viewed at present. The first part deals with knowledge-based systems and comprises eleven contributions from leading experts in the field. The emphasis here is primarily on the use of artificial intelligence, expert systems and other knowledge-based systems for fault detection and reliability. The second part is devoted to fault detection of technological systems and comprises thirteen contributions dealing with applications of fault detection techniques to various technological systems such as gas networks, electric power systems, nuclear reactors and assembly cells. The third part of the proceedings, which consists of seven contributions, treats robust, fault tolerant and intelligent controllers and covers methodological issues as well as several applications ranging from nuclear power plants to industrial robots to steel grinding. The fourth part treats fault tolerant digital techniques and comprises five contributions. Two papers, one on reactor noise analysis, the other on reactor control system design, are indexed separately. (author)

  18. A reliable simultaneous representation of seismic hazard and of ground shaking recurrence

    Science.gov (United States)

    Peresan, A.; Panza, G. F.; Magrin, A.; Vaccari, F.

    2015-12-01

    Different earthquake hazard maps may be appropriate for different purposes - such as emergency management, insurance and engineering design. Accounting for the lower occurrence rate of larger sporadic earthquakes may allow to formulate cost-effective policies in some specific applications, provided that statistically sound recurrence estimates are used, which is not typically the case of PSHA (Probabilistic Seismic Hazard Assessment). We illustrate the procedure to associate the expected ground motions from Neo-deterministic Seismic Hazard Assessment (NDSHA) to an estimate of their recurrence. Neo-deterministic refers to a scenario-based approach, which allows for the construction of a broad range of earthquake scenarios via full waveforms modeling. From the synthetic seismograms the estimates of peak ground acceleration, velocity and displacement, or any other parameter relevant to seismic engineering, can be extracted. NDSHA, in its standard form, defines the hazard computed from a wide set of scenario earthquakes (including the largest deterministically or historically defined credible earthquake, MCE) and it does not supply the frequency of occurrence of the expected ground shaking. A recent enhanced variant of NDSHA that reliably accounts for recurrence has been developed and it is applied to the Italian territory. The characterization of the frequency-magnitude relation can be performed by any statistically sound method supported by data (e.g. multi-scale seismicity model), so that a recurrence estimate is associated to each of the pertinent sources. In this way a standard NDSHA map of ground shaking is obtained simultaneously with the map of the corresponding recurrences. The introduction of recurrence estimates in NDSHA naturally allows for the generation of ground shaking maps at specified return periods. This permits a straightforward comparison between NDSHA and PSHA maps.

  19. RELIABILITY OF THE DETECTION OF THE BARYON ACOUSTIC PEAK

    International Nuclear Information System (INIS)

    MartInez, Vicent J.; Arnalte-Mur, Pablo; De la Cruz, Pablo; Saar, Enn; Tempel, Elmo; Pons-BorderIa, MarIa Jesus; Paredes, Silvestre; Fernandez-Soto, Alberto

    2009-01-01

    The correlation function of the distribution of matter in the universe shows, at large scales, baryon acoustic oscillations, which were imprinted prior to recombination. This feature was first detected in the correlation function of the luminous red galaxies of the Sloan Digital Sky Survey (SDSS). Recently, the final release (DR7) of the SDSS has been made available, and the useful volume is about two times bigger than in the old sample. We present here, for the first time, the redshift-space correlation function of this sample at large scales together with that for one shallower, but denser volume-limited subsample drawn from the Two-Degree Field Redshift Survey. We test the reliability of the detection of the acoustic peak at about 100 h -1 Mpc and the behavior of the correlation function at larger scales by means of careful estimation of errors. We confirm the presence of the peak in the latest data although broader than in previous detections.

  20. Reliability analysis for the quench detection in the LHC machine

    CERN Document Server

    Denz, R; Vergara-Fernández, A

    2002-01-01

    The Large Hadron Collider (LHC) will incorporate a large amount of superconducting elements that require protection in case of a quench. Key elements in the quench protection system are the electronic quench detectors. Their reliability will have an important impact on the down time as well as on the operational cost of the collider. The expected rates of both false and missed quenches have been computed for several redundant detection schemes. The developed model takes account of the maintainability of the system to optimise the frequency of foreseen checks, and evaluate their influence on the performance of different detection topologies. Seen the uncertainty of the failure rate of the components combined with the LHC tunnel environment, the study has been completed with a sensitivity analysis of the results. The chosen detection scheme and the maintainability strategy for each detector family are given.

  1. A signal processing framework for simultaneous detection of multiple environmental contaminants

    International Nuclear Information System (INIS)

    Chakraborty, Subhadeep; Mench, Matthew M; Manahan, Michael P

    2013-01-01

    The possibility of large-scale attacks using chemical warfare agents (CWAs) has exposed the critical need for fundamental research enabling the reliable, unambiguous and early detection of trace CWAs and toxic industrial chemicals. This paper presents a unique approach for the identification and classification of simultaneously present multiple environmental contaminants by perturbing an electrochemical (EC) sensor with an oscillating potential for the extraction of statistically rich information from the current response. The dynamic response, being a function of the degree and mechanism of contamination, is then processed with a symbolic dynamic filter for the extraction of representative patterns, which are then classified using a trained neural network. The approach presented in this paper promises to extend the sensing power and sensitivity of these EC sensors by augmenting and complementing sensor technology with state-of-the-art embedded real-time signal processing capabilities. (paper)

  2. Design of a multiplex PCR assay for the simultaneous detection and confirmation of Neisseria gonorrhoeae.

    LENUS (Irish Health Repository)

    O'Callaghan, Isabelle

    2010-05-01

    To improve the detection of Neisseria gonorrhoeae by designing a multiplex PCR assay using two N gonorrhoeae-specific genes as targets, thereby providing detection and confirmation of a positive result simultaneously.

  3. A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species.

    Science.gov (United States)

    Radhika, M; Saugata, Majumder; Murali, H S; Batra, H V

    2014-01-01

    Salmonella enterica and Shigella species are commonly associated with food and water borne infections leading to gastrointestinal diseases. The present work was undertaken to develop a sensitive and reliable PCR based detection system for simultaneous detection of Salmonella enterica and Shigella at species level. For this the conserved regions of specific genes namely ipaH1, ipaH, wbgZ, wzy and invA were targeted for detection of Shigella genus, S. flexneri, S. sonnei, S. boydii and Salmonella enterica respectively along with an internal amplification control (IAC). The results showed that twenty Salmonella and eleven Shigella spp., were accurately identified by the assay without showing non-specificity against closely related other Enterobacteriaceae organisms and also against other pathogens. Further evaluation of multiplex PCR was undertaken on 50 natural samples of chicken, eggs and poultry litter and results compared with conventional culture isolation and identification procedure. The multiplex PCR identified the presence of Salmonella and Shigella strains with a short pre-enrichment step of 5 h in peptone water and the same samples were processed by conventional procedures for comparison. Therefore, this reported multiplex PCR can serve as an alternative to the tedious time-consuming procedure of culture and identification in food safety laboratories.

  4. The reliability of a maximal isometric hip strength and simultaneous surface EMG screening protocol in elite, junior rugby league athletes.

    Science.gov (United States)

    Charlton, Paula C; Mentiplay, Benjamin F; Grimaldi, Alison; Pua, Yong-Hao; Clark, Ross A

    2017-02-01

    Firstly to describe the reliability of assessing maximal isometric strength of the hip abductor and adductor musculature using a hand held dynamometry (HHD) protocol with simultaneous wireless surface electromyographic (sEMG) evaluation of the gluteus medius (GM) and adductor longus (AL). Secondly, to describe the correlation between isometric strength recorded with the HHD protocol and a laboratory standard isokinetic device. Reliability and correlational study. A sample of 24 elite, male, junior, rugby league athletes, age 16-20 years participated in repeated HHD and isometric Kin-Com (KC) strength testing with simultaneous sEMG assessment, on average (range) 6 (5-7) days apart by a single assessor. Strength tests included; unilateral hip abduction (ABD) and adduction (ADD) and bilateral ADD assessed with squeeze (SQ) tests in 0 and 45° of hip flexion. HHD demonstrated good to excellent inter-session reliability for all outcome measures (ICC (2,1) =0.76-0.91) and good to excellent association with the laboratory reference KC (ICC (2,1) =0.80-0.88). Whilst intra-session, inter-trial reliability of EMG activation and co-activation outcome measures ranged from moderate to excellent (ICC (2,1) =0.70-0.94), inter-session reliability was poor (all ICC (2,1) Isometric strength testing of the hip ABD and ADD musculature using HHD may be measured reliably in elite, junior rugby league athletes. Due to the poor inter-session reliability of sEMG measures, it is not recommended for athlete screening purposes if using the techniques implemented in this study. Copyright © 2016 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.

  5. Novel real-time polymerase chain reaction assay for simultaneous detection of recurrent fusion genes in acute myeloid leukemia.

    Science.gov (United States)

    Dolz, Sandra; Barragán, Eva; Fuster, Óscar; Llop, Marta; Cervera, José; Such, Esperanza; De Juan, Inmaculada; Palanca, Sarai; Murria, Rosa; Bolufer, Pascual; Luna, Irene; Gómez, Inés; López, María; Ibáñez, Mariam; Sanz, Miguel A

    2013-09-01

    The recent World Health Organization classification recognizes different subtypes of acute myeloid leukemia (AML) according to the presence of several recurrent genetic abnormalities. Detection of these abnormalities and other molecular changes is of increasing interest because it contributes to a refined diagnosis and prognostic assessment in AML and enables monitoring of minimal residual disease. These genetic abnormalities can be detected using single RT-PCR, although the screening is still labor intensive and costly. We have developed a novel real-time RT-PCR assay to simultaneously detect 15 AML-associated rearrangements that is a simple and easily applicable method for use in clinical diagnostic laboratories. This method showed 100% specificity and sensitivity (95% confidence interval, 91% to 100% and 92% to 100%, respectively). The procedure was validated in a series of 105 patients with AML. The method confirmed all translocations detected using standard cytogenetics and fluorescence in situ hybridization and some additional undetected rearrangements. Two patients demonstrated two molecular rearrangements simultaneously, with BCR-ABL1 implicated in both, in addition to RUNX1-MECOM in one patient and PML-RARA in another. In conclusion, this novel real-time RT-PCR assay for simultaneous detection of multiple AML-associated fusion genes is a versatile and sensitive method for reliable screening of recurrent rearrangements in AML. Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

  6. Simultaneous detection of respiratory syncytial virus types A and B ...

    African Journals Online (AJOL)

    Tharwat Ezzat Deraz

    2012-02-28

    Feb 28, 2012 ... Abstract Background: Respiratory syncytial virus (RSV) types A and B and influenza A and B cause about .... plied Science, Mannheim, Germany; Cat. ..... detection of human rhinoviruses, paramyxoviruses, corona viruses, and ...

  7. Simultaneous Determination of Different Anions in Milk Samples Using Ion Chromatography with Conductivity Detection

    Directory of Open Access Journals (Sweden)

    Gülçin Gümüş Yılmaz

    2016-12-01

    Full Text Available The description of a simple method for simultaneous determination of chloride, nitrate, sulfate, iodide, phosphate, thiocyanate, perchlorate, and orotic acid in milk samples was outlined. The method involves the use of dialysis cassettes for matrix elimination, followed by ion chromatography on a high capacity anion exchange column with suppressed conductivity detection. The novelty of dialysis process was that it did not need any chemical and organic solvent for elimination of macromolecules such as fat, carbohydrates and proteins from milk samples. External standard calibration curves for these analytes were linear with great correlation coefficients. The relative standard deviations of analyte concentrations were acceptable both inter-day and intra-day evaluations. Under optimized conditions, the limit of detection (Signal-to-Noise ratio = 3 for chloride, phosphate, thiocyanate, perchlorate, iodide, nitrate, sulfate, and orotate was found to be 0.012, 0.112, 0.140, 0.280, 0.312, 0.516, 0.520, and 0.840 mg L−1, respectively. Significant results were obtained for various spiked milk samples with % recovery in the range of 93.88 - 109.75 %. The proposed method was successfully applied to milk samples collected from Istanbul markets. The advantages of the method described herein are reagent-free, simple, and reliable.

  8. Simultaneous Detection of Brown Rot- and Soft Rot-Causing Bacterial Pathogens from Potato Tubers Through Multiplex PCR.

    Science.gov (United States)

    Ranjan, R K; Singh, Dinesh; Baranwal, V K

    2016-11-01

    Ralstonia solanacearum (Smith) Yabuuchi et al. and Erwinia carotovora subsp. carotovora (Jones) Bergey et al. (Pectobacterium carotovorum subsp. carotovorum) are the two major bacterial pathogens of potato causing brown rot (wilt) and soft rot diseases, respectively, in the field and during storage. Reliable and early detection of these pathogens are keys to avoid occurrence of these diseases in potato crops and reduce yield loss. In the present study, multiplex polymerase chain reaction (PCR) protocol was developed for simultaneous detection of R. solanacearum and E. carotovora subsp. carotovora from potato tubers. A set of oligos targeting the pectatelyase (pel) gene of E. carotovora subsp. carotovora and the universal primers based on 16S r RNA gene of R. solanacearum were used. The standardized multiplex PCR protocol could detect R. solanacearum and E. carotovora subsp. carotovora up to 0.01 and 1.0 ng of genomic DNA, respectively. The protocol was further validated on 96 stored potato tuber samples, collected from different potato-growing states of India, viz. Uttarakhand, Odisha, Meghalaya and Delhi. 53.1 % tuber samples were positive for R. solanacearum, and 15.1 % of samples were positive for E. carotovora subsp. carotovora, and both the pathogens were positive in 26.0 % samples when BIO-PCR was used. This method offers sensitive, specific, reliable and fast detection of two major bacterial pathogens from potato tubers simultaneously, particularly pathogen-free seed certification in large scale.

  9. Simultaneous detection of Apple mosaic virus in cultivated hazelnuts ...

    African Journals Online (AJOL)

    The most economically damaging ilarvirus affecting hazelnut on a worldwide scale is the related apple mosaic virus (ApMV). Attempts were made to isolate the virus RNA from hazelnut tissues using different extraction methods. The most suitable extraction method that could detect the virus occurring naturally in hazelnut by ...

  10. Simultaneous parameter and tolerance optimization of structures via probability-interval mixed reliability model

    DEFF Research Database (Denmark)

    Luo, Yangjun; Wu, Xiaoxiang; Zhou, Mingdong

    2015-01-01

    Both structural sizes and dimensional tolerances strongly influence the manufacturing cost and the functional performance of a practical product. This paper presents an optimization method to simultaneously find the optimal combination of structural sizes and dimensional tolerances. Based...... transformed into their equivalent formulations by using the performance measure approach. The optimization problem is then solved with the sequential approximate programming. Meanwhile, a numerically stable algorithm based on the trust region method is proposed to efficiently update the target performance...

  11. Reverse Transcription Polymerase Chain Reaction-based System for Simultaneous Detection of Multiple Lily-infecting Viruses

    Directory of Open Access Journals (Sweden)

    Ji Yeon Kwon

    2013-09-01

    Full Text Available A detection system based on a multiplex reverse transcription (RT polymerase chain reaction (PCR was developed to simultaneously identify multiple viruses in the lily plant. The most common viruses infecting lily plants are the cucumber mosaic virus (CMV, lily mottle virus (LMoV, lily symptomless virus (LSV. Leaf samples were collected at lily-cultivation facilities located in the Kangwon province of Korea and used to evaluate the detection system. Simplex and multiplex RT-PCR were performed using virus-specific primers to detect single-or mixed viral infections in lily plants. Our results demonstrate the selective detection of 3 different viruses (CMV, LMoV and LSV by using specific primers as well as the potential of simultaneously detecting 2 or 3 different viruses in lily plants with mixed infections. Three sets of primers for each target virus, and one set of internal control primers were used to evaluate the detection system for efficiency, reliability, and reproducibility.

  12. Simultaneous mass detection for direct inlet mass spectrometry

    International Nuclear Information System (INIS)

    Gordon, R.L.

    1979-05-01

    The evolution of analytical techniques for application in trace analysis has led to interest in practical methods for real-time monitoring. Direct inlet mass spectrometry (DIMS) has been the subject of considerable activity in recent years. A DIMS instrument is described which consists of an inlet system designed to permit particles entrained in the inlet air stream to strike a hot, oxidized rhenium filament which serves as a surface ionization source. A mass analyzer and detection system then permits identification of the elemental composition of particulates which strike the filament

  13. Detecting binary black holes with efficient and reliable templates

    International Nuclear Information System (INIS)

    Damour, T.; Iyer, B.R.; Sathyaprakash, B.S.

    2001-01-01

    Detecting binary black holes in interferometer data requires an accurate knowledge of the orbital phase evolution of the system. From the point of view of data analysis one also needs fast algorithms to compute the templates that will be employed in searching for black hole binaries. Recently, there has been progress on both these fronts: On one hand, re-summation techniques have made it possible to accelerate the convergence of poorly convergent asymptotic post-Newtonian series and derive waveforms beyond the conventional adiabatic approximation. We now have a waveform model that extends beyond the inspiral regime into the plunge phase followed by the quasi-normal mode ringing. On the other hand, explicit Fourier domain waveforms have been derived that make the generation of waveforms fast enough so as not to be a burden on the computational resources required in filtering the detector data. These new developments should make it possible to efficiently and reliably search for black hole binaries in data from first interferometers. (author)

  14. Reliability

    OpenAIRE

    Condon, David; Revelle, William

    2017-01-01

    Separating the signal in a test from the irrelevant noise is a challenge for all measurement. Low test reliability limits test validity, attenuates important relationships, and can lead to regression artifacts. Multiple approaches to the assessment and improvement of reliability are discussed. The advantages and disadvantages of several different approaches to reliability are considered. Practical advice on how to assess reliability using open source software is provided.

  15. A one-step multiplex RT-PCR assay for simultaneous detection of four viruses that infect peach.

    Science.gov (United States)

    Yu, Y; Zhao, Z; Jiang, D; Wu, Z; Li, S

    2013-10-01

    A multiplex reverse transcription polymerase chain reaction (mRT-PCR) assay was developed to enable the simultaneous detection and differentiation of four viruses that infect peach, namely Apple chlorotic leaf spot virus (ACLSV), Cherry green ring mottle virus (CGRMV), Prunus necrotic ringspot virus (PNRSV) and Apricot pseudo-chlorotic leaf spot virus (APCLSV). In this study, four pairs of primers, one specific for each virus, were designed; the corresponding PCR products were 632, 439, 346 and 282 bp in length for ACLSV, CGRMV, PNRSV and APCLSV, respectively, and the fragments could be distinguished clearly by agarose gel electrophoresis. The sensitivity and specificity of the method were tested using individual RT-PCR and enzyme-linked immunosorbent assay (ELISA), and the identity of the RT-PCR amplification products was also confirmed by DNA sequencing. The results of RT-PCR and ELISA, along with batch detection using samples collected from peach orchards, revealed that this rapid and simple technique is an effective way to identify the four viruses simultaneously. The mRT-PCR assay described in this study was developed for the simultaneous detection of four peach viruses from infected peach samples is reliable and sensitive. In contrast to conventional uniplex RT-PCR, mRT-PCR is more efficient, reducing costs, time and handling when testing large numbers of samples. This rapid and simple method is useful for large-scale surveys of viruses that infect peach. © 2013 The Society for Applied Microbiology.

  16. Reliable simultaneous zymographic method of characterization of cellulolytic enzymes from fungal cellulase complex.

    Science.gov (United States)

    Dojnov, Biljana; Grujić, Marica; Vujčić, Zoran

    2015-08-01

    A method for zymographic detection of specific cellulases in a complex (endocellulase, exocellulase, and cellobiase) from crude fermentation extracts, after a single electrophoretic separation, is described in this paper. Cellulases were printed onto a membrane and, subsequently, substrate gel. Cellobiase isoforms were detected on the membrane using esculine as substrate, endocellulase isoforms on substrate gel with copolymerized carboxymethyl cellulose (CMC), while exocellulase isoforms were detected in electrophoresis gel with 4-methylumbelliferyl-β-d-cellobioside (MUC). This can be a useful additional tool for monitoring and control of fungal cellulase production in industrial processes and fundamental research, screening for particular cellulase producers, or testing of new lignocellulose substrates. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Simultaneous detection of Hepatitis B surface antigen and its antibody by radioimmunoassay

    International Nuclear Information System (INIS)

    Crouzat-Reynes, Gerard; Perigois, Francois; Lecureuil, Michel; Lejeune, Bernard

    1981-01-01

    The authors describe an original radioimmunoassay which allows the simultaneous detection of hepatitis B surface antigen and its antibody in a biological sample. Antigen and antibody are indiscriminately detected in a first step and then distinguished in a second step using the same reagents [fr

  18. Electrophoresis microchip with integrated waveguides for simultaneous native UV fluorescence and absorbance detection

    DEFF Research Database (Denmark)

    Ohlsson, Pelle Daniel; Sala, Olga Ordeig; Mogensen, Klaus Bo

    2009-01-01

    Simultaneous label-free detection of UV absorbance and native UV-excited fluorescence in an electrophoresis microchip is presented. UV transparent integrated waveguides launch light at a wavelength of 254 nm from a mercury lamp along the length of a 1-mm. long detection cell. Transmitted UV light...

  19. Reliability of leak detection systems in light water reactors

    International Nuclear Information System (INIS)

    Kupperman, D.S.

    1987-01-01

    US Nuclear Regulatory Commission Guide 1.45 recommends the use of at least three different detection methods in reactors to detect leakage. Monitoring of both sump-flow and airborne particulate radioactivity is recommended. A third method can involve either monitoring of condensate flow rate from air coolers or monitoring of airborne gaseous radioactivity. Although the methods currently used for leak detection reflect the state of the art, other techniques may be developed and used. Since the recommendations of Regulatory Guide 1.45 are not mandatory, the technical specifications for 74 operating plants have been reviewed to determine the types of leak detection methods employed. In addition, Licensee Event Report (LER) Compilations from June 1985 to June 1986 have been reviewed to help establish actual capabilities for detecting leaks and determining their source. Work at Argonne National Laboratory has demonstrated that improvements in leak detection, location, and sizing are possible with advanced acoustic leak detection technology

  20. Using multiple PCR and CE with chemiluminescence detection for simultaneous qualitative and quantitative analysis of genetically modified organism.

    Science.gov (United States)

    Guo, Longhua; Qiu, Bin; Chi, Yuwu; Chen, Guonan

    2008-09-01

    In this paper, an ultrasensitive CE-CL detection system coupled with a novel double-on-column coaxial flow detection interface was developed for the detection of PCR products. A reliable procedure based on this system had been demonstrated for qualitative and quantitative analysis of genetically modified organism-the detection of Roundup Ready Soy (RRS) samples was presented as an example. The promoter, terminator, function and two reference genes of RRS were amplified with multiplex PCR simultaneously. After that, the multiplex PCR products were labeled with acridinium ester at the 5'-terminal through an amino modification and then analyzed by the proposed CE-CL system. Reproducibility of analysis times and peak heights for the CE-CL analysis were determined to be better than 0.91 and 3.07% (RSD, n=15), respectively, for three consecutive days. It was shown that this method could accurately and qualitatively detect RRS standards and the simulative samples. The evaluation in terms of quantitative analysis of RRS provided by this new method was confirmed by comparing our assay results with those of the standard real-time quantitative PCR (RT-QPCR) using SYBR Green I dyes. The results showed a good coherence between the two methods. This approach demonstrated the possibility for accurate qualitative and quantitative detection of GM plants in a single run.

  1. Simultaneous Detection of Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes at a Very Low Level Using Simultaneous Enrichment Broth and Multichannel SPR Biosensor.

    Science.gov (United States)

    Zhang, Xiaoguang; Tsuji, Sachiko; Kitaoka, Hayato; Kobayashi, Hiroshi; Tamai, Mitsuru; Honjoh, Ken-Ichi; Miyamoto, Takahisa

    2017-10-01

    Detection of foodborne pathogens at very low levels is still a challenge. A custom-built multichannel surface plasmon resonance (SPR) biosensor and simultaneous enrichment broth (SEB) were used to develop a simultaneous detection method for 3 important foodborne pathogens, Escherichia coli O157:H7 (O157:H7), Salmonella enteritidis, and Listeria monocytogenes, at a very low level. These 3 foodborne pathogens at a very low level (14, 6, and 28 CFU/25 g (mL) for O157:H7, S. enteritidis, and L. monocytogenes, respectively) were inoculated in SEB and incubated at 37 ˚C for 24 h. Sample prepared from the simultaneous enrichment culture was analyzed using the multichannel SPR biosensor and sensor chip immobilized with polyclonal antibodies specific to each of the target pathogens. O157:H7, S. enteritidis, and L. monocytogenes in chicken were detected simultaneously at an inoculum dose of 14, 6, and 28 CFU/25 g, respectively. Our method using a custom-built multichannel SPR biosensor and enrichment in SEB is expected as a rapid and simultaneous detection method for low levels of O157:H7, S. enteritidis, and L. monocytogenes in food. Our method is expected as a rapid and simultaneous detection method for pathogens at very low levels. It has great potential for safety control of food and microbiological detection applications. © 2017 Institute of Food Technologists®.

  2. Detecting and Remembering Simultaneous Pictures in a Rapid Serial Visual Presentation

    Science.gov (United States)

    Potter, Mary C.; Fox, Laura F.

    2009-01-01

    Viewers can easily spot a target picture in a rapid serial visual presentation (RSVP), but can they do so if more than 1 picture is presented simultaneously? Up to 4 pictures were presented on each RSVP frame, for 240 to 720 ms/frame. In a detection task, the target was verbally specified before each trial (e.g., "man with violin"); in a…

  3. Reliability of semiquantitative 18F-FDG PET parameters derived from simultaneous brain PET/MRI: A feasibility study

    International Nuclear Information System (INIS)

    Jena, Amarnath; Taneja, Sangeeta; Goel, Reema; Renjen, Pushpendranath; Negi, Pradeep

    2014-01-01

    Purpose: Simultaneous brain PET/MRI faces an important issue of validation of accurate MRI based attenuation correction (AC) method for precise quantitation of brain PET data unlike in PET/CT systems where the use of standard, validated CT based AC is routinely available. The aim of this study was to investigate the feasibility of evaluation of semiquantitative 18 F-FDG PET parameters derived from simultaneous brain PET/MRI using ultrashort echo time (UTE) sequences for AC and to assess their agreement with those obtained from PET/CT examination. Methods: Sixteen patients (age range 18–73 years; mean age 49.43 (19.3) years; 13 men 3 women) underwent simultaneous brain PET/MRI followed immediately by PET/CT. Quantitative analysis of brain PET images obtained from both studies was undertaken using Scenium v.1 brain analysis software package. Twenty ROIs for various brain regions were system generated and 6 semiquantitative parameters including maximum standardized uptake value (SUV max), SUV mean, minimum SUV (SUV min), minimum standard deviation (SD min), maximum SD (SD max) and SD from mean were calculated for both sets of PET data for each patient. Intra-class correlation coefficients (ICCs) were determined to assess agreement between the various semiquantitative parameters for the two PET data sets. Results: Intra-class co-relation between the two PET data sets for SUV max, SUV mean and SD max was highly significant (p < 0.00) for all the 20 predefined brain regions with ICC > 0.9. SD from mean was also found to be statistically significant for all the predefined brain regions with ICC > 0.8. However, SUV max and SUV mean values obtained from PET/MRI were significantly lower compared to those of PET/CT for all the predefined brain regions. Conclusion: PET quantitation accuracy using the MRI based UTE sequences for AC in simultaneous brain PET/MRI is reliable in a clinical setting, being similar to that obtained using PET/CT

  4. PV Systems Reliability Final Technical Report: Ground Fault Detection

    Energy Technology Data Exchange (ETDEWEB)

    Lavrova, Olga [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Flicker, Jack David [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Johnson, Jay [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2016-01-01

    We have examined ground faults in PhotoVoltaic (PV) arrays and the efficacy of fuse, current detection (RCD), current sense monitoring/relays (CSM), isolation/insulation (Riso) monitoring, and Ground Fault Detection and Isolation (GFID) using simulations based on a Simulation Program with Integrated Circuit Emphasis SPICE ground fault circuit model, experimental ground faults installed on real arrays, and theoretical equations.

  5. Simultaneous fluorescent detection of multiple metal ions based on the DNAzymes and graphene oxide.

    Science.gov (United States)

    Yun, Wen; Wu, Hong; Liu, Xingyan; Fu, Min; Jiang, Jiaolai; Du, Yunfeng; Yang, Lizhu; Huang, Yu

    2017-09-15

    A novel fluorescent detection strategy for simultaneous detection of Cu 2+ , Pb 2+ and Mg 2+ based on DNAzyme branched junction structure with three kinds of DNAzymes and graphene oxide (GO) was presented. Three fluorophores labeled DNA sequences consisted with enzyme-strand (E-DNA) and substrate strand (S-DNA) were annealed to form DNAzyme branched junction structure. In the presence of target metal ion, the DNAzyme was activated to cleave the fluorophore labeled S-DNA. The S-DNA fragments were released and adsorbed onto GO surface to quench the fluorescent signal. The detection limit was calculated to be 1 nM for Cu 2+ , 200 nM for Mg 2+ , and 0.3 nM for Pb 2+ , respectively. This strategy was successfully used for simultaneous detection of Cu 2+ , Mg 2+ and Pb 2+ in human serum. Moreover, it had potential application for simultaneous detection of multiple metal ions in environmental and biological samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Using detection dogs to conduct simultaneous surveys of northern spotted (Strix occidentalis caurina and barred owls (Strix varia.

    Directory of Open Access Journals (Sweden)

    Samuel K Wasser

    Full Text Available State and federal actions to conserve northern spotted owl (Strix occidentalis caurina habitat are largely initiated by establishing habitat occupancy. Northern spotted owl occupancy is typically assessed by eliciting their response to simulated conspecific vocalizations. However, proximity of barred owls (Strix varia-a significant threat to northern spotted owls-can suppress northern spotted owl responsiveness to vocalization surveys and hence their probability of detection. We developed a survey method to simultaneously detect both species that does not require vocalization. Detection dogs (Canis familiaris located owl pellets accumulated under roost sites, within search areas selected using habitat association maps. We compared success of detection dog surveys to vocalization surveys slightly modified from the U.S. Fish and Wildlife Service's Draft 2010 Survey Protocol. Seventeen 2 km × 2 km polygons were each surveyed multiple times in an area where northern spotted owls were known to nest prior to 1997 and barred owl density was thought to be low. Mitochondrial DNA was used to confirm species from pellets detected by dogs. Spotted owl and barred owl detection probabilities were significantly higher for dog than vocalization surveys. For spotted owls, this difference increased with number of site visits. Cumulative detection probabilities of northern spotted owls were 29% after session 1, 62% after session 2, and 87% after session 3 for dog surveys, compared to 25% after session 1, increasing to 59% by session 6 for vocalization surveys. Mean detection probability for barred owls was 20.1% for dog surveys and 7.3% for vocal surveys. Results suggest that detection dog surveys can complement vocalization surveys by providing a reliable method for establishing occupancy of both northern spotted and barred owl without requiring owl vocalization. This helps meet objectives of Recovery Actions 24 and 25 of the Revised Recovery Plan for the

  7. A gas sensor array for the simultaneous detection of multiple VOCs.

    Science.gov (United States)

    Zhang, Yumin; Zhao, Jianhong; Du, Tengfei; Zhu, Zhongqi; Zhang, Jin; Liu, Qingju

    2017-05-16

    Air quality around the globe is declining and public health is seriously threatened by indoor air pollution. Typically, indoor air pollutants are composed of a series of volatile organic compounds (VOCs) that are generally harmful to the human body, especially VOCs with low molecular weights (less than 100 Da). Moreover, in some situations, more than one type of VOC is present; thus, a device that can detect one or more VOCs simultaneously would be most beneficial. Here, we synthesized a sensor array with 4 units to detect 4 VOCs: acetone (unit 1), benzene (unit 2), methanol (unit 3) and formaldehyde (unit 4) simultaneously. All units were simultaneously exposed to 2.5 ppm of all four VOCs. The sensitivity of unit 1 was 14.67 for acetone and less than 2.54 for the other VOCs. The sensitivities of units 2, 3 and 4 to benzene, methanol and formaldehyde were 2 18.64, 20.98 and 17.26, respectively, and less than 4.01 for the other VOCs. These results indicated that the sensor array exhibited good selectivity and could be used for the real-time monitoring of indoor air quality. Thus, this device will be useful in situations requiring the simultaneous detection of multiple VOCs.

  8. Speech detection in noise and spatial unmasking in children with simultaneous versus sequential bilateral cochlear implants.

    Science.gov (United States)

    Chadha, Neil K; Papsin, Blake C; Jiwani, Salima; Gordon, Karen A

    2011-09-01

    To measure speech detection in noise performance for children with bilateral cochlear implants (BiCI), to compare performance in children with simultaneous implant versus those with sequential implant, and to compare performance to normal-hearing children. Prospective cohort study. Tertiary academic pediatric center. Children with early-onset bilateral deafness and 2-year BiCI experience, comprising the "sequential" group (>2 yr interimplantation delay, n = 12) and "simultaneous group" (no interimplantation delay, n = 10) and normal-hearing controls (n = 8). Thresholds to speech detection (at 0-degree azimuth) were measured with noise at 0-degree azimuth or ± 90-degree azimuth. Spatial unmasking (SU) as the noise condition changed from 0-degree azimuth to ± 90-degree azimuth and binaural summation advantage (BSA) of 2 over 1 CI. Speech detection in noise was significantly poorer than controls for both BiCI groups (p simultaneous group approached levels found in normal controls (7.2 ± 0.6 versus 8.6 ± 0.6 dB, p > 0.05) and was significantly better than that in the sequential group (3.9 ± 0.4 dB, p simultaneous group but, in the sequential group, was significantly better when noise was moved to the second rather than the first implanted ear (4.8 ± 0.5 versus 3.0 ± 0.4 dB, p sequential group's second rather than first CI. Children with simultaneously implanted BiCI demonstrated an advantage over children with sequential implant by using spatial cues to improve speech detection in noise.

  9. Simultaneous detection of pH changes and histamine release from oxyntic glands in isolated stomach.

    Science.gov (United States)

    Bitziou, Eleni; O'Hare, Danny; Patel, Bhavik Anil

    2008-11-15

    Real-time simultaneous detection of changes in pH and levels of histamine over the oxyntic glands of guinea pig stomach have been investigated. An iridium oxide pH microelectrode was used in a potentiometric mode to record the pH decrease associated with acid secretion when the sensor approached the isolated tissue. A boron-doped diamond (BDD) microelectrode was used in an amperometric mode to detect histamine when the electrode was placed over the tissue. Both sensors provided stable and reproducible responses that were qualitatively consistent with the signaling mechanism for acid secretion at the stomach. Simultaneous measurements in the presence of pharmacological treatments produced significant variations in the signals obtained by both sensors. As the H2 receptor antagonist cimetidine was perfused to the tissue, histamine levels increased that produced an increase in the signal of the BDD electrode whereas the pH sensor recorded a decrease in acid secretion as expected. Addition of acetylcholine (ACh) stimulated additional acid secretion detected with the pH microelectrode whereas the BDD sensor recorded the histamine levels decreasing significantly. This result shows that the primary influence of ACh is directly on the parietal cell receptors rather then the ECL cell receptors of the oxyntic glands. These results highlight the power of this simultaneous detection technique in the monitoring and diagnosis of physiological significant signaling mechanisms and pathways.

  10. Epigallocatechin Gallate-Modified Graphite Paste Electrode for Simultaneous Detection of Redox-Active Biomolecules

    Directory of Open Access Journals (Sweden)

    Hashwin V. S. Ganesh

    2017-12-01

    Full Text Available In this study, simultaneous electrochemical detection of ascorbic acid (AA, dopamine (DA, and uric acid (UA was performed using a modified graphite paste electrode (MGPE with epigallocatechin gallate (EGCG and green tea (GT powder. It was shown that the anodic peak current increased in comparison with that of the graphite paste electrode (GPE in the cyclic voltammograms. The optimal pH for simultaneous determination of a quaternary mixture of AA–DA–UA was determined to be pH 2. The anodic peak potentials for a mixture containing AA–DA–UA were well separated from each other. The catalytic peak currents obtained at the surface of the MGPE/EGCG were linearly dependent on the AA, DA, and UA concentrations up to 23, 14, and 14 µM, respectively. The detection limits for AA, DA, and UA were 190, 90, and 70 nM, respectively. The analytical performance of this sensor has been evaluated for simultaneous detection of AA, DA, and UA in real samples. Finally, a modified electrode was prepared using GT and used for simultaneous determination of AA, DA, and UA. Based on the results, MPGE/GT showed two oxidation peaks at 0.43 and 0.6 V for DA and UA, respectively, without any oxidation peak for AA. The calibration curves at the surface of MGPE/GT were linear up to 14 µM with a detection limit of 0.18 and 0.33 µM for DA and UA, respectively. MGPEs provide a promising platform for the future development of sensors for multiplexed electrochemical detection of clinically important analytes.

  11. Continuous wave protocol for simultaneous polarization and optical detection of P1-center electron spin resonance

    Science.gov (United States)

    Kamp, E. J.; Carvajal, B.; Samarth, N.

    2018-01-01

    The ready optical detection and manipulation of bright nitrogen vacancy center spins in diamond plays a key role in contemporary quantum information science and quantum metrology. Other optically dark defects such as substitutional nitrogen atoms (`P1 centers') could also become potentially useful in this context if they could be as easily optically detected and manipulated. We develop a relatively straightforward continuous wave protocol that takes advantage of the dipolar coupling between nitrogen vacancy and P1 centers in type 1b diamond to detect and polarize the dark P1 spins. By combining mutual spin flip transitions with radio frequency driving, we demonstrate the simultaneous optical polarization and detection of the electron spin resonance of the P1 center. This technique should be applicable to detecting and manipulating a broad range of dark spin populations that couple to the nitrogen vacancy center via dipolar fields, allowing for quantum metrology using these spin populations.

  12. Objective Methods for Reliable Detection of Concealed Depression

    Directory of Open Access Journals (Sweden)

    Cynthia eSolomon

    2015-04-01

    Full Text Available Recent research has shown that it is possible to automatically detect clinical depression from audio-visual recordings. Before considering integration in a clinical pathway, a key question that must be asked is whether such systems can be easily fooled. This work explores the potential of acoustic features to detect clinical depression in adults both when acting normally and when asked to conceal their depression. Nine adults diagnosed with mild to moderate depression as per the Beck Depression Inventory (BDI-II and Patient Health Questionnaire (PHQ-9 were asked a series of questions and to read a excerpt from a novel aloud under two different experimental conditions. In one, participants were asked to act naturally and in the other, to suppress anything that they felt would be indicative of their depression. Acoustic features were then extracted from this data and analysed using paired t-tests to determine any statistically significant differences between healthy and depressed participants. Most features that were found to be significantly different during normal behaviour remained so during concealed behaviour. In leave-one-subject-out automatic classification studies of the 9 depressed subjects and 8 matched healthy controls, an 88% classification accuracy and 89% sensitivity was achieved. Results remained relatively robust during concealed behaviour, with classifiers trained on only non-concealed data achieving 81% detection accuracy and 75% sensitivity when tested on concealed data. These results indicate there is good potential to build deception-proof automatic depression monitoring systems.

  13. Research Note The reliability of a field test kit for the detection and ...

    African Journals Online (AJOL)

    Research Note The reliability of a field test kit for the detection and the persistence of ... Open Access DOWNLOAD FULL TEXT ... The objectives were to test a field kit for practicality and reliability, to assess the spread of the bacteria among ...

  14. Terahertz-wave differential detection based on simultaneous dual-wavelength up-conversion

    Directory of Open Access Journals (Sweden)

    Yuma Takida

    2017-03-01

    Full Text Available We report a terahertz (THz-wave differential detection based on simultaneous dual-wavelength up-conversion in a nonlinear optical MgO:LiNbO3 crystal with optical and electronic THz-wave sources. The broadband parametric gain and noncollinear phase-matching of MgO:LiNbO3 provide efficient conversion from superposed THz waves to spatially distributed near-infrared (NIR beams to function as a dispersive THz-wave spectrometer without any additional dispersive element. We show that the μW-level THz waves from two independent sources, a 0.78-THz injection-seeded THz-wave parametric generator (is-TPG and a 1.14-THz resonant tunneling diode (RTD, are simultaneously up-converted to two NIR waves and then detected with two NIR photodetectors. By applying a balanced detection scheme to this dual-frequency detection, we demonstrate THz-wave differential imaging of maltose and polyethylene pellets in the transmission geometry. This dual-wavelength detection is applicable to more than three frequencies and broadband THz-wave radiation for real-time THz-wave spectroscopic detection and imaging.

  15. Three-dimensional fluorescent microscopy via simultaneous illumination and detection at multiple planes.

    Science.gov (United States)

    Ma, Qian; Khademhosseinieh, Bahar; Huang, Eric; Qian, Haoliang; Bakowski, Malina A; Troemel, Emily R; Liu, Zhaowei

    2016-08-16

    The conventional optical microscope is an inherently two-dimensional (2D) imaging tool. The objective lens, eyepiece and image sensor are all designed to capture light emitted from a 2D 'object plane'. Existing technologies, such as confocal or light sheet fluorescence microscopy have to utilize mechanical scanning, a time-multiplexing process, to capture a 3D image. In this paper, we present a 3D optical microscopy method based upon simultaneously illuminating and detecting multiple focal planes. This is implemented by adding two diffractive optical elements to modify the illumination and detection optics. We demonstrate that the image quality of this technique is comparable to conventional light sheet fluorescent microscopy with the advantage of the simultaneous imaging of multiple axial planes and reduced number of scans required to image the whole sample volume.

  16. Towards Reliable Evaluation of Anomaly-Based Intrusion Detection Performance

    Science.gov (United States)

    Viswanathan, Arun

    2012-01-01

    This report describes the results of research into the effects of environment-induced noise on the evaluation process for anomaly detectors in the cyber security domain. This research was conducted during a 10-week summer internship program from the 19th of August, 2012 to the 23rd of August, 2012 at the Jet Propulsion Laboratory in Pasadena, California. The research performed lies within the larger context of the Los Angeles Department of Water and Power (LADWP) Smart Grid cyber security project, a Department of Energy (DoE) funded effort involving the Jet Propulsion Laboratory, California Institute of Technology and the University of Southern California/ Information Sciences Institute. The results of the present effort constitute an important contribution towards building more rigorous evaluation paradigms for anomaly-based intrusion detectors in complex cyber physical systems such as the Smart Grid. Anomaly detection is a key strategy for cyber intrusion detection and operates by identifying deviations from profiles of nominal behavior and are thus conceptually appealing for detecting "novel" attacks. Evaluating the performance of such a detector requires assessing: (a) how well it captures the model of nominal behavior, and (b) how well it detects attacks (deviations from normality). Current evaluation methods produce results that give insufficient insight into the operation of a detector, inevitably resulting in a significantly poor characterization of a detectors performance. In this work, we first describe a preliminary taxonomy of key evaluation constructs that are necessary for establishing rigor in the evaluation regime of an anomaly detector. We then focus on clarifying the impact of the operational environment on the manifestation of attacks in monitored data. We show how dynamic and evolving environments can introduce high variability into the data stream perturbing detector performance. Prior research has focused on understanding the impact of this

  17. Comparison of Two Suspension Arrays for Simultaneous Detection of Five Biothreat Bacterial in Powder Samples

    Directory of Open Access Journals (Sweden)

    Yu Yang

    2012-01-01

    Full Text Available We have developed novel Bio-Plex assays for simultaneous detection of Bacillus anthracis, Yersinia pestis, Brucella spp., Francisella tularensis, and Burkholderia pseudomallei. Universal primers were used to amplify highly conserved region located within the 16S rRNA amplicon, followed by hybridized to pathogen-specific probes for identification of these five organisms. The other assay is based on multiplex PCR to simultaneously amplify five species-specific pathogen identification-targeted regions unique to individual pathogen. Both of the two arrays are validated to be flexible and sensitive for simultaneous detection of bioterrorism bacteria. However, universal primer PCR-based array could not identify Bacillus anthracis, Yersinia pestis, and Brucella spp. at the species level because of the high conservation of 16S rDNA of the same genus. The two suspension arrays can be utilized to detect Bacillus anthracis sterne spore and Yersinia pestis EV76 from mimic “write powder” samples, they also proved that the suspension array system will be valuable tools for diagnosis of bacterial biothreat agents in environmental samples.

  18. Simultaneous determination of ethamsylate, tramadol and lidocaine in human urine by capillary electrophoresis with electrochemiluminescence detection.

    Science.gov (United States)

    Li, Jianguo; Ju, Huangxian

    2006-09-01

    Ethamsylate, tramadol and lidocaine, partly excreted by the kidney, are generally used as hemostatic, analgesic and local anesthetic in surgery. We developed a simple and sensitive method for their simultaneous monitoring in human urine based on CE coupled with electrochemiluminescence detection by end-column mode. Under optimized conditions the proposed method yielded linear ranges from 5.0 x 10(-8) to 5.0 x 10(-5), 1.0 x 10(-7) to 1.0 x 10(-4) and 1.0 x 10(-7) to 1.0 x 10(-4) M with LODs of 8.0 x 10(-9) M (36 amol), 1.6 x 10(-8) M (72 amol) and 1.0 x 10(-8) M (45 amol) (S/N = 3) for ethamsylate, tramadol and lidocaine, respectively. The RSD for their simultaneous detection at 1.0 x 10(-6) M was 2.1, 2.8 and 3.2% (n = 7), respectively. For practical application an extraction step with ethyl acetate at pH 11 was performed to eliminate the influence of the sample ionic strength. The recoveries of ethamsylate, tramadol and lidocaine at different levels in human urine were between 87 and 95%. This method was used for simultaneous detection of ethamsylate, tramadol and lidocaine in clinic urine samples from two medicated patients. It was valuable in clinical and biochemical laboratories for monitoring these drugs for various purposes.

  19. A Turbidity Test Based Centrifugal Microfluidics Diagnostic System for Simultaneous Detection of HBV, HCV, and CMV

    Directory of Open Access Journals (Sweden)

    Hung-Cheng Chang

    2015-01-01

    Full Text Available This paper presents a LAMP- (loop-mediated isothermal amplification- based lab-on-disk optical system that allows the simultaneous detection of hepatitis B virus, hepatitis C virus, and cytomegalovirus. The various flow stages are controlled in the proposed system using different balance among centrifugal pumping, Coriolis pumping, and the capillary force. We have implemented a servo system for positioning and speed control for the heating and centrifugal pumping. We have also successfully employed a polymer light-emitting diode section for turbidity detection. The easy-to-use one-click system can perform diagnostics in less than 1 hour.

  20. Rapid Simultaneous Amplification and Detection of the MBR/JH Chromosomal Translocation by Fluorescence Melting Curve Analysis

    Science.gov (United States)

    Bohling, Sandra D.; King, Thomas C.; Wittwer, Carl T.; Elenitoba-Johnson, Kojo S. J.

    1999-01-01

    Polymerase chain reaction (PCR) amplification and product analysis for the detection of chromosomal translocations, such as the t(14;18), has traditionally been a two-step process. PCR product detection has generally entailed gel electrophoresis and/or hybridization or sequencing for confirmation of assay specificity. Using a microvolume fluorimeter integrated with a thermal cycler and a PCR-compatible double-stranded DNA (dsDNA) binding fluorescent dye (SYBR Green I), we investigated the feasibility of simultaneous thermal amplification and detection of MBR/JH translocation products by fluorescence melting curve analysis. We analyzed DNA from 30 cases of lymphoproliferative disorders comprising 19 cases of previously documented MBR/JH-positive follicle center lymphoma and 11 reactive lymphadenopathies. The samples were coded and analyzed blindly for the presence of MBR/JH translocations by fluorescence melting curve analysis. We also performed dilutional assays using the MBR/JH-positive cell line SUDHL-6. Multiplex PCR for MBR/JH and β-globin was used to simultaneously assess sample adequacy. All (100%) of the 19 cases previously determined to be MBR/JH positive by conventional PCR analysis showed a characteristic sharp decrease in fluorescence at ∼90°C by melting curve analysis after amplification. Fluorescence melting peaks obtained by plotting the negative derivative of fluorescence over temperature (−dF/dT) versus temperature (T) showed melting temperatures (Tm) at 88.85 ± 1.15°C. In addition, multiplex assays using both MBR/JH and β-globin primers yielded easily distinguishable fluorescence melting peaks at ∼90°C and 81.2°C, respectively. Dilutional assays revealed that fluorescence melting curve analysis was more sensitive than conventional PCR and agarose gel electrophoresis with ultraviolet transillumination by as much as 100-fold. Simultaneous amplification and fluorescence melting curve analysis is a simple, reliable, and sensitive method

  1. Bedside ultrasound reliability in locating catheter and detecting complications

    Directory of Open Access Journals (Sweden)

    Payman Moharamzadeh

    2016-10-01

    Full Text Available Introduction: Central venous catheterization is one of the most common medical procedures and is associated with such complications as misplacement and pneumothorax. Chest X-ray is among good ways for evaluation of these complications. However, due to patient’s excessive exposure to radiation, time consumption and low diagnostic value in detecting pneumothorax in the supine patient, the present study intends to examine bedside ultrasound diagnostic value in locating tip of the catheter and pneumothorax. Materials and methods: In the present cross-sectional study, all referred patients requiring central venous catheterization were examined. Central venous catheterization was performed by a trained emergency medicine specialist, and the location of catheter and the presence of pneumothorax were examined and compared using two modalities of ultrasound and x-ray (as the reference standard. Sensitivity, specificity, and positive and negative predicting values were reported. Results: A total of 200 non-trauma patients were included in the study (58% men. Cohen’s Kappa consistency coefficients for catheterization and diagnosis of pneumothorax were found as 0.49 (95% CI: 0.43-0.55, 0.89 (P<0.001, (95% CI: 97.8-100, respectively. Also, ultrasound sensitivity and specificity in diagnosing pneumothorax were 75% (95% CI: 35.6-95.5, and 100% (95% CI: 97.6-100, respectively. Conclusion: The present study results showed low diagnostic value of ultrasound in determining catheter location and in detecting pneumothorax. With knowledge of previous studies, the search still on this field.   Keywords: Central venous catheterization; complications; bedside ultrasound; radiography;

  2. Reliability considerations of electronics components for the deep underwater muon and neutrino detection system

    International Nuclear Information System (INIS)

    Leskovar, B.

    1980-02-01

    The reliability of some electronics components for the Deep Underwater Muon and Neutrino Detection (DUMAND) System is discussed. An introductory overview of engineering concepts and technique for reliability assessment is given. Component reliability is discussed in the contest of major factors causing failures, particularly with respect to physical and chemical causes, process technology and testing, and screening procedures. Failure rates are presented for discrete devices and for integrated circuits as well as for basic electronics components. Furthermore, the military reliability specifications and standards for semiconductor devices are reviewed

  3. A reliable and validated LC-MS/MS method for the simultaneous quantification of 4 cannabinoids in 40 consumer products.

    Directory of Open Access Journals (Sweden)

    Qingfang Meng

    Full Text Available In the past 50 years, Cannabis sativa (C. sativa has gone from a substance essentially prohibited worldwide to one that is gaining acceptance both culturally and legally in many countries for medicinal and recreational use. As additional jurisdictions legalize Cannabis products and the variety and complexity of these products surpass the classical dried plant material, appropriate methods for measuring the biologically active constituents is paramount to ensure safety and regulatory compliance. While there are numerous active compounds in C. sativa the primary cannabinoids of regulatory and safety concern are (--Δ⁹-tetrahydrocannabinol (THC, cannabidiol (CBD, and their respective acidic forms THCA-A and CBDA. Using the US Food and Drug Administration (FDA bioanalytical method validation guidelines we developed a sensitive, selective, and accurate method for the simultaneous analysis CBD, CBDA, THC, and THCA-A in oils and THC & CBD in more complex matrices. This HPLC-MS/MS method was simple and reliable using standard sample dilution and homogenization, an isocratic chromatographic separation, and a triple quadrupole mass spectrometer. The lower limit of quantification (LLOQ for analytes was 0.195 ng/mL over a 0.195-50.0 ng/mL range of quantification with a coefficient of correlation of >0.99. Average intra-day and inter-day accuracies were 94.2-112.7% and 97.2-110.9%, respectively. This method was used to quantify CBD, CBDA, THC, and THCA-A in 40 commercial hemp products representing a variety of matrices including oils, plant materials, and creams/cosmetics. All products tested met the federal regulatory restrictions on THC content in Canada (1,000 (an oil-based product. Overall, the method proved amenable to the analysis of various commercial products including oils, creams, and plant material and may be diagnostically indicative of adulteration with non-hemp C. sativa, specialized hemp cultivars, or unique manufacturing methods.

  4. SIMULTANEOUS MULTI-BAND DETECTION OF LOW SURFACE BRIGHTNESS GALAXIES WITH MARKOVIAN MODELING

    International Nuclear Information System (INIS)

    Vollmer, B.; Bonnarel, F.; Louys, M.; Perret, B.; Petremand, M.; Lavigne, F.; Collet, Ch.; Van Driel, W.; Sabatini, S.; MacArthur, L. A.

    2013-01-01

    We present to the astronomical community an algorithm for the detection of low surface brightness (LSB) galaxies in images, called MARSIAA (MARkovian Software for Image Analysis in Astronomy), which is based on multi-scale Markovian modeling. MARSIAA can be applied simultaneously to different bands. It segments an image into a user-defined number of classes, according to their surface brightness and surroundings—typically, one or two classes contain the LSB structures. We have developed an algorithm, called DetectLSB, which allows the efficient identification of LSB galaxies from among the candidate sources selected by MARSIAA. The application of the method to two and three bands simultaneously was tested on simulated images. Based on our tests, we are confident that we can detect LSB galaxies down to a central surface brightness level of only 1.5 times the standard deviation from the mean pixel value in the image background. To assess the robustness of our method, the method was applied to a set of 18 B- and I-band images (covering 1.3 deg 2 in total) of the Virgo Cluster to which Sabatini et al. previously applied a matched-filter dwarf LSB galaxy search algorithm. We have detected all 20 objects from the Sabatini et al. catalog which we could classify by eye as bona fide LSB galaxies. Our method has also detected four additional Virgo Cluster LSB galaxy candidates undetected by Sabatini et al. To further assess the completeness of the results of our method, both MARSIAA, SExtractor, and DetectLSB were applied to search for (1) mock Virgo LSB galaxies inserted into a set of deep Next Generation Virgo Survey (NGVS) gri-band subimages and (2) Virgo LSB galaxies identified by eye in a full set of NGVS square degree gri images. MARSIAA/DetectLSB recovered ∼20% more mock LSB galaxies and ∼40% more LSB galaxies identified by eye than SExtractor/DetectLSB. With a 90% fraction of false positives from an entirely unsupervised pipeline, a completeness of 90% is

  5. Development of recombinant antigen array for simultaneous detection of viral antibodies.

    Directory of Open Access Journals (Sweden)

    Yi Liu

    Full Text Available Protein microarrays have been developed to study antibody reactivity against a large number of antigens, demonstrating extensive perspective for clinical application. We developed a viral antigen array by spotting four recombinant antigens and synthetic peptide, including glycoprotein G of herpes simplex virus (HSV type 1 and 2, phosphoprotein 150 of cytomegalovirus (CMV, Rubella virus (RV core plus glycoprotein E1 and E2 as well as a E1 peptide with the optimal concentrations on activated glass slides to simultaneously detect IgG and IgM against HSV1, HSV2, CMV and RV in clinical specimens of sera and cerebrospinal fluids (CSFs. The positive reference sera were initially used to measure the sensitivity and specificity of the array with the optimal conditions. Then clinical specimens of 144 sera and 93 CSFs were tested for IgG and IgM antibodies directed against HSV1, HSV2, CMV and RV by the antigen array. Specificity of the antigen array for viral antibodies detection was satisfying compared to commercial ELISA kits but sensitivity of the array varied relying on quality and antigenic epitopes of the spotting antigens. In short, the recombinant antigen array has potential to simultaneous detect multiple viral antibodies using minute amount (3 µl of samples, which holds the particularly advantage to detect viral antibodies in clinical CSFs being suspicious of neonatal meningitis and encephalitis.

  6. The Reliability and Effectiveness of a Radar-Based Animal Detection System

    Science.gov (United States)

    2017-09-22

    This document contains data on the reliability and effectiveness of an animal detection system along U.S. Hwy 95 near Bonners Ferry, Idaho. The system uses a Doppler radar to detect large mammals (e.g., deer and elk) when they approach the highway. T...

  7. The Reliability and Effectiveness of a Radar-Based Animal Detection System

    Science.gov (United States)

    2017-09-01

    This document contains data on the reliability and effectiveness of an animal detection system along U.S. Hwy 95 near Bonners Ferry, Idaho. The system uses a Doppler radar to detect large mammals (e.g., deer and elk) when they approach the highway. T...

  8. Simultaneous and rapid detection of six different mycotoxins using an immunochip.

    Science.gov (United States)

    Wang, Ying; Liu, Nan; Ning, Baoan; Liu, Ming; Lv, Zhiqiang; Sun, Zhiyong; Peng, Yuan; Chen, Cuicui; Li, Junwen; Gao, Zhixian

    2012-04-15

    Mycotoxins are highly toxic contaminants in food, animal feed, and commodities. The study has developed an immunochip for quantifying the concentrations of six mycotoxins: aflatoxin B1, aflatoxin M1, deoxynivalenol, ochratoxin A, T-2 toxin, and zearalenone, which were added to drinking water. The complete antigens (Ags) of the mycotoxins were contact printed and immobilized onto agarose-modified glass slides with 12 physically isolated subarrays, based on the reaction of both diffusion and covalent bond. The optimal concentration of each antigen and antibody (Ab) was obtained using an Ag-Ab immunoassay. Based on the indirect competitive immunoassay for the simultaneous detection of six mycotoxins in one single chip, six standard curves with good logistic correlation (R(2)>0.97) were respectively plotted. The working ranges (0.04-1.69, 0.45-3.90, 20.20-69.23, 35.68-363.18, 0.11-1.81, and 0.08-7.47 ng/mL, respectively) were calculated, as well as the median inhibitory concentrations (0.31±0.04, 1.49±0.21, 34.54±1.30, 134.06±11.75, 0.49±0.05, and 1.54±0.22 ng/mL, respectively), when six mycotoxins were detected simultaneously. Finally, the recovery rates in drinking water generally ranged from 80% to 120% on the same chip, with an intra-assay coefficient of variation lower than 15%. We successfully established an immunochip for simultaneous detection of six mycotoxins within 4h, with advantages of using minimal samples and being visually semiquantitative with our naked eyes. In summary, the method could be developed on one single chip for detecting multiple contaminants in actual samples. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Simultaneous Whole-Brain Segmentation and White Matter Lesion Detection Using Contrast-Adaptive Probabilistic Models

    DEFF Research Database (Denmark)

    Puonti, Oula; Van Leemput, Koen

    2016-01-01

    In this paper we propose a new generative model for simultaneous brain parcellation and white matter lesion segmentation from multi-contrast magnetic resonance images. The method combines an existing whole-brain segmentation technique with a novel spatial lesion model based on a convolutional...... restricted Boltzmann machine. Unlike current state-of-the-art lesion detection techniques based on discriminative modeling, the proposed method is not tuned to one specific scanner or imaging protocol, and simultaneously segments dozens of neuroanatomical structures. Experiments on a public benchmark dataset...... in multiple sclerosis indicate that the method’s lesion segmentation accuracy compares well to that of the current state-of-the-art in the field, while additionally providing robust whole-brain segmentations....

  10. Simultaneous detection of iodine and iodide on boron doped diamond electrodes.

    Science.gov (United States)

    Fierro, Stéphane; Comninellis, Christos; Einaga, Yasuaki

    2013-01-15

    Individual and simultaneous electrochemical detection of iodide and iodine has been performed via cyclic voltammetry on boron doped diamond (BDD) electrodes in a 1M NaClO(4) (pH 8) solution, representative of typical environmental water conditions. It is feasible to compute accurate calibration curve for both compounds using cyclic voltammetry measurements by determining the peak current intensities as a function of the concentration. A lower detection limit of about 20 μM was obtained for iodide and 10 μM for iodine. Based on the comparison between the peak current intensities reported during the oxidation of KI, it is probable that iodide (I(-)) is first oxidized in a single step to yield iodine (I(2)). The latter is further oxidized to obtain IO(3)(-). This technique, however, did not allow for a reasonably accurate detection of iodate (IO(3)(-)) on a BDD electrode. Copyright © 2012 Elsevier B.V. All rights reserved.

  11. Rapid, highly sensitive and highly specific gene detection by combining enzymatic amplification and DNA chip detection simultaneously

    Directory of Open Access Journals (Sweden)

    Koji Hashimoto

    2016-05-01

    Full Text Available We have developed a novel gene detection method based on the loop-mediated isothermal amplification (LAMP reaction and the DNA dissociation reaction on the same DNA chip surface to achieve a lower detection limit, broader dynamic range and faster detection time than are attainable with a conventional DNA chip. Both FAM- and thiol-labeled DNA probe bound to the complementary sequence accompanying Dabcyl was immobilized on the gold surface via Au/thiol bond. The LAMP reaction was carried out on the DNA probe fixed gold surface. At first, Dabcyl molecules quenched the FAM fluorescence. According to the LAMP reaction, the complementary sequence with Dabcyl was competitively reacted with the amplified targeted sequence. As a result, the FAM fluorescence increased owing to dissociation of the complementary sequence from the DNA probe. The simultaneous reaction of LAMP and DNA chip detection was achieved, and 103 copies of the targeted gene were detected within an hour by measuring fluorescence intensity of the DNA probe. Keywords: Biosensor, DNA chip, Loop-mediated isothermal amplification (LAMP, Fluorescence detection, Gold substrate, Au/thiol bond

  12. Ultrasensitive Detection of RNA and DNA Viruses Simultaneously Using Duplex UNDP-PCR Assay

    Science.gov (United States)

    Wang, Zengguo; Zhang, Xiujuan; Zhao, Xiaomin; Du, Qian; Chang, Lingling; Tong, Dewen

    2015-01-01

    Mixed infection of multiple viruses is common in modern intensive pig rearing. However, there are no methods available to detect DNA and RNA viruses in the same reaction system in preclinical level. In this study, we aimed to develop a duplex ultrasensitive nanoparticle DNA probe-based PCR assay (duplex UNDP-PCR) that was able to simultaneously detect DNA and RNA viruses in the same reaction system. PCV2 and TGEV are selected as representatives of the two different types of viruses. PCV2 DNA and TGEV RNA were simultaneously released from the serum sample by boiling with lysis buffer, then magnetic beads and gold nanoparticles coated with single and/or duplex specific probes for TGEV and PCV2 were added to form a sandwich-like complex with nucleic acids released from viruses. After magnetic separation, DNA barcodes specific for PCV2 and TGEV were eluted using DTT and characterized by specific PCR assay for specific DNA barcodes subsequently. The duplex UNDP-PCR showed similar sensitivity as that of single UNDP-PCR and was able to detect 20 copies each of PCV2 and TGEV in the serum, showing approximately 250-fold more sensitivity than conventional duplex PCR/RT-PCR assays. No cross-reaction was observed with other viruses. The positive detection rate of single MMPs- and duplex MMPs-based duplex UNDP-PCR was identical, with 29.6% for PCV2, 9.3% for TGEV and 3.7% for PCV2 and TGEV mixed infection. This duplex UNDP-PCR assay could detect TGEV (RNA virus) and PCV2 (DNA virus) from large-scale serum samples simultaneously without the need for DNA/RNA extraction, purification and reverse transcription of RNA, and showed a significantly increased positive detection rate for PCV2 (29%) and TGEV (11.7%) preclinical infection than conventional duplex PCR/RT-PCR. Therefore, the established duplex UNDP-PCR is a rapid and economical detection method, exhibiting high sensitivity, specificity and reproducibility. PMID:26544710

  13. Ultrasensitive Detection of RNA and DNA Viruses Simultaneously Using Duplex UNDP-PCR Assay.

    Directory of Open Access Journals (Sweden)

    Yong Huang

    Full Text Available Mixed infection of multiple viruses is common in modern intensive pig rearing. However, there are no methods available to detect DNA and RNA viruses in the same reaction system in preclinical level. In this study, we aimed to develop a duplex ultrasensitive nanoparticle DNA probe-based PCR assay (duplex UNDP-PCR that was able to simultaneously detect DNA and RNA viruses in the same reaction system. PCV2 and TGEV are selected as representatives of the two different types of viruses. PCV2 DNA and TGEV RNA were simultaneously released from the serum sample by boiling with lysis buffer, then magnetic beads and gold nanoparticles coated with single and/or duplex specific probes for TGEV and PCV2 were added to form a sandwich-like complex with nucleic acids released from viruses. After magnetic separation, DNA barcodes specific for PCV2 and TGEV were eluted using DTT and characterized by specific PCR assay for specific DNA barcodes subsequently. The duplex UNDP-PCR showed similar sensitivity as that of single UNDP-PCR and was able to detect 20 copies each of PCV2 and TGEV in the serum, showing approximately 250-fold more sensitivity than conventional duplex PCR/RT-PCR assays. No cross-reaction was observed with other viruses. The positive detection rate of single MMPs- and duplex MMPs-based duplex UNDP-PCR was identical, with 29.6% for PCV2, 9.3% for TGEV and 3.7% for PCV2 and TGEV mixed infection. This duplex UNDP-PCR assay could detect TGEV (RNA virus and PCV2 (DNA virus from large-scale serum samples simultaneously without the need for DNA/RNA extraction, purification and reverse transcription of RNA, and showed a significantly increased positive detection rate for PCV2 (29% and TGEV (11.7% preclinical infection than conventional duplex PCR/RT-PCR. Therefore, the established duplex UNDP-PCR is a rapid and economical detection method, exhibiting high sensitivity, specificity and reproducibility.

  14. Ultrasensitive Detection of RNA and DNA Viruses Simultaneously Using Duplex UNDP-PCR Assay.

    Science.gov (United States)

    Huang, Yong; Xing, Na; Wang, Zengguo; Zhang, Xiujuan; Zhao, Xiaomin; Du, Qian; Chang, Lingling; Tong, Dewen

    2015-01-01

    Mixed infection of multiple viruses is common in modern intensive pig rearing. However, there are no methods available to detect DNA and RNA viruses in the same reaction system in preclinical level. In this study, we aimed to develop a duplex ultrasensitive nanoparticle DNA probe-based PCR assay (duplex UNDP-PCR) that was able to simultaneously detect DNA and RNA viruses in the same reaction system. PCV2 and TGEV are selected as representatives of the two different types of viruses. PCV2 DNA and TGEV RNA were simultaneously released from the serum sample by boiling with lysis buffer, then magnetic beads and gold nanoparticles coated with single and/or duplex specific probes for TGEV and PCV2 were added to form a sandwich-like complex with nucleic acids released from viruses. After magnetic separation, DNA barcodes specific for PCV2 and TGEV were eluted using DTT and characterized by specific PCR assay for specific DNA barcodes subsequently. The duplex UNDP-PCR showed similar sensitivity as that of single UNDP-PCR and was able to detect 20 copies each of PCV2 and TGEV in the serum, showing approximately 250-fold more sensitivity than conventional duplex PCR/RT-PCR assays. No cross-reaction was observed with other viruses. The positive detection rate of single MMPs- and duplex MMPs-based duplex UNDP-PCR was identical, with 29.6% for PCV2, 9.3% for TGEV and 3.7% for PCV2 and TGEV mixed infection. This duplex UNDP-PCR assay could detect TGEV (RNA virus) and PCV2 (DNA virus) from large-scale serum samples simultaneously without the need for DNA/RNA extraction, purification and reverse transcription of RNA, and showed a significantly increased positive detection rate for PCV2 (29%) and TGEV (11.7%) preclinical infection than conventional duplex PCR/RT-PCR. Therefore, the established duplex UNDP-PCR is a rapid and economical detection method, exhibiting high sensitivity, specificity and reproducibility.

  15. Detection of focal epileptic activity using combined simultaneous electroencephalogram-functional MRI

    International Nuclear Information System (INIS)

    Zhang Zhiqiang; Lu Guangming; Tian Lei; Sun Kanjian; Tan Qifu; Zhu Jianguo; Nie Cong; Hao Shaowei; Jiang Li; Liu Yijun

    2007-01-01

    Objective: To observe the brain activation of interictal epiletiform discharges (IEDs) and to localize the epileptogenic foci of epilepsy. Methods: The electroencephalogram (EEG) and functional MRI data of 12 focal epileptic patients were acquired using a combination of EEG and functional MRI simultaneously. The IEDs onset time detected with EEG were set as the time parameters in an event- related paradigm of functional MRI analysis. The spatial and temporal characters of IEDs activation were analyzed in detail. In order to confirm the consistency of this method, all patients were scanned repeatedly and the results were correlated with clinical evaluation. Results: Of the 12 patients, valid data from EEG- fMRI were obtained from 10 patients in a total of 18 sessions. Compared with the structural foci, the epileptic foci localization results of eleven sessions were good, five sessions were fairly good, and two sessions were poor. The results obtained from six patients in two separate sessions were concordant, respectively. Moreover, thalamic activation was detected in ten sessions, cerebellar activation was detected in all sessions, and the deactivation was found in the default mode loci in nine sessions. Conclusion: The method of performing EEG and fMRI simultaneously can potentially be a useful tool in epilepsy research. (authors)

  16. Simultaneous Detection of Five Pathogens from Cerebrospinal Fluid Specimens Using Luminex Technology

    Directory of Open Access Journals (Sweden)

    Linfu Zhou

    2016-02-01

    Full Text Available Early diagnosis and treatment are crucial for the outcome of central nervous system (CNS infections. In this study, we developed a multiplex PCR-Luminex assay for the simultaneous detection of five major pathogens, including Mycobacterium tuberculosis, Cryptococcus neoformans, Streptococcus pneumoniae, and herpes simplex virus types 1 and 2, which frequently cause CNS infections. Through the hybridization reaction between multiplex PCR-amplified targets and oligonucleotide “anti-TAG” sequences, we found that the PCR-Luminex assay could detect as low as 101–102 copies of synthetic pathogen DNAs. Furthermore, 163 cerebrospinal fluid (CSF specimens from patients with suspected CNS infections were used to evaluate the efficiency of this multiplex PCR-Luminex method. Compared with Ziehl-Neelsen stain, this assay showed a high diagnostic accuracy for tuberculosis meningitis (sensitivity, 90.7% and specificity, 99.1%. For cryptococcal meningitis, the sensitivity and specificity were 92% and 97.1%, respectively, compared with the May Grunwald Giemsa (MGG stain. For herpes simplex virus types 1 and 2 encephalitis, the sensitivities were 80.8% and 100%, and the specificities were 94.2% and 99%, respectively, compared with Enzyme Linked Immunosorbent Assay (ELISA assays. Taken together, this multiplex PCR-Luminex assay showed potential efficiency for the simultaneous detection of five pathogens and may be a promising supplement to conventional methods for diagnosing CNS infections.

  17. Simultaneous detection of ascorbic acid, uric acid and homovanillic acid at copper modified electrode

    International Nuclear Information System (INIS)

    Selvaraju, T.; Ramaraj, R.

    2007-01-01

    The copper was deposited on glassy carbon (GC) and indium tin oxide (ITO) electrodes by electrochemical method. The copper structures on electrode were characterized by atomic force microscope, X-ray diffractometeric pattern and differential pulse voltammetric studies. Optimal conditions for uniform growth of copper structures on the electrode were established. Voltammetric sensor was fabricated using the copper deposited GC electrode for the simultaneous detection and determination of uric acid (UA) and homovanillic acid (HVA) in the presence of excess concentrations of ascorbic acid (AA). The voltammetric signals due to AA and UA oxidation were well separated with a potential difference of 400 mV and AA did not interfere with the measurement of UA and HVA at the GC/Cu electrode. Linear calibration curves were obtained in the concentration range 1-40 μM for AA and 20-50 μM for UA at physiological pH and a detection limit of 10 nM of UA in the presence of 10-fold excess concentrations of AA was achieved. The simultaneous detection of submicromolar concentrations of AA, UA and HVA was achieved at the GC/Cu electrode. The practical utility of the present GC/Cu modified electrode was demonstrated by measuring the AA content in Vitamin C tablet, UA content in human urine and blood serum samples with satisfactory results

  18. Simultaneous detection of lactate and glucose by integrated printed circuit board based array sensing chip

    Energy Technology Data Exchange (ETDEWEB)

    Li, Xuelian [Institute for Clean Energy and Advanced Materials, Southwest University, Chongqing 400715 (China); School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Zang, Jianfeng [Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708 (United States); Liu, Yingshuai; Lu, Zhisong [Institute for Clean Energy and Advanced Materials, Southwest University, Chongqing 400715 (China); Li, Qing, E-mail: Qli@swu.edu.cn [School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Li, Chang Ming, E-mail: ecmli@swu.edu.cn [Institute for Clean Energy and Advanced Materials, Southwest University, Chongqing 400715 (China)

    2013-04-10

    Highlights: ► An integrated printed circuit board (PCB) based array sensing chip was developed. ► Simultaneous detection of lactate and glucose in serum has been demonstrated. ► The array electronic biochip has high signal to noise ratio and high sensitivity. ► Additional electrodes were designed on the chip to correct interferences. -- Abstract: An integrated printed circuit board (PCB) based array sensing chip was developed to simultaneously detect lactate and glucose in mouse serum. The novelty of the chip relies on a concept demonstration of inexpensive high-throughput electronic biochip, a chip design for high signal to noise ratio and high sensitivity by construction of positively charged chitosan/redox polymer Polyvinylimidazole-Os (PVI-Os)/carbon nanotube (CNT) composite sensing platform, in which the positively charged chitosan/PVI-Os is mediator and electrostatically immobilizes the negatively charged enzyme, while CNTs function as an essential cross-linker to network PVI-Os and chitosan due to its negative charged nature. Additional electrodes on the chip with the same sensing layer but without enzymes were prepared to correct the interferences for high specificity. Low detection limits of 0.6 μM and 5 μM were achieved for lactate and glucose, respectively. This work could be extended to inexpensive array sensing chips with high sensitivity, good specificity and high reproducibility for various sensor applications.

  19. Simultaneous aptasensor for multiplex pathogenic bacteria detection based on multicolor upconversion nanoparticles labels.

    Science.gov (United States)

    Wu, Shijia; Duan, Nuo; Shi, Zhao; Fang, Congcong; Wang, Zhouping

    2014-03-18

    A highly sensitive and specific multiplex method for the simultaneous detection of three pathogenic bacteria was fabricated using multicolor upconversion nanoparticles (UCNPs) as luminescence labels coupled with aptamers as the molecular recognition elements. Multicolor UCNPs were synthesized via doping with various rare-earth ions to obtain well-separated emission peaks. The aptamer sequences were selected using the systematic evolution of ligands by exponential enrichment (SELEX) strategy for Staphylococcus aureus, Vibrio parahemolyticus, and Salmonella typhimurium. When applied in this method, aptamers can be used for the specific recognition of the bacteria from complex mixtures, including those found in real food matrixes. Aptamers and multicolor UCNPs were employed to selectively capture and simultaneously quantify the three target bacteria on the basis of the independent peaks. Under optimal conditions, the correlation between the concentration of three bacteria and the luminescence signal was found to be linear from 50-10(6) cfu mL(-1). Improved by the magnetic separation and concentration effect of Fe3O4 magnetic nanoparticles, the limits of detection of the developed method were found to be 25, 10, and 15 cfu mL(-1) for S. aureus, V. parahemolyticus, and S. typhimurium, respectively. The capability of the bioassay in real food samples was also investigated, and the results were consistent with experimental results obtained from plate-counting methods. This proposed method for the detection of various pathogenic bacteria based on multicolor UCNPs has great potential in the application of food safety and multiplex nanosensors.

  20. Simultaneous capture and sequential detection of two malarial biomarkers on magnetic microparticles.

    Science.gov (United States)

    Markwalter, Christine F; Ricks, Keersten M; Bitting, Anna L; Mudenda, Lwiindi; Wright, David W

    2016-12-01

    We have developed a rapid magnetic microparticle-based detection strategy for malarial biomarkers Plasmodium lactate dehydrogenase (pLDH) and Plasmodium falciparum histidine-rich protein II (PfHRPII). In this assay, magnetic particles functionalized with antibodies specific for pLDH and PfHRPII as well as detection antibodies with distinct enzymes for each biomarker are added to parasitized lysed blood samples. Sandwich complexes for pLDH and PfHRPII form on the surface of the magnetic beads, which are washed and sequentially re-suspended in detection enzyme substrate for each antigen. The developed simultaneous capture and sequential detection (SCSD) assay detects both biomarkers in samples as low as 2.0parasites/µl, an order of magnitude below commercially available ELISA kits, has a total incubation time of 35min, and was found to be reproducible between users over time. This assay provides a simple and efficient alternative to traditional 96-well plate ELISAs, which take 5-8h to complete and are limited to one analyte. Further, the modularity of the magnetic bead-based SCSD ELISA format could serve as a platform for application to other diseases for which multi-biomarker detection is advantageous. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  1. A reliable and validated LC-MS/MS method for the simultaneous quantification of 4 cannabinoids in 40 consumer products.

    Science.gov (United States)

    Meng, Qingfang; Buchanan, Beth; Zuccolo, Jonathan; Poulin, Mathieu-Marc; Gabriele, Joseph; Baranowski, David Charles

    2018-01-01

    In the past 50 years, Cannabis sativa (C. sativa) has gone from a substance essentially prohibited worldwide to one that is gaining acceptance both culturally and legally in many countries for medicinal and recreational use. As additional jurisdictions legalize Cannabis products and the variety and complexity of these products surpass the classical dried plant material, appropriate methods for measuring the biologically active constituents is paramount to ensure safety and regulatory compliance. While there are numerous active compounds in C. sativa the primary cannabinoids of regulatory and safety concern are (-)-Δ⁹-tetrahydrocannabinol (THC), cannabidiol (CBD), and their respective acidic forms THCA-A and CBDA. Using the US Food and Drug Administration (FDA) bioanalytical method validation guidelines we developed a sensitive, selective, and accurate method for the simultaneous analysis CBD, CBDA, THC, and THCA-A in oils and THC & CBD in more complex matrices. This HPLC-MS/MS method was simple and reliable using standard sample dilution and homogenization, an isocratic chromatographic separation, and a triple quadrupole mass spectrometer. The lower limit of quantification (LLOQ) for analytes was 0.195 ng/mL over a 0.195-50.0 ng/mL range of quantification with a coefficient of correlation of >0.99. Average intra-day and inter-day accuracies were 94.2-112.7% and 97.2-110.9%, respectively. This method was used to quantify CBD, CBDA, THC, and THCA-A in 40 commercial hemp products representing a variety of matrices including oils, plant materials, and creams/cosmetics. All products tested met the federal regulatory restrictions on THC content in Canada (CBD, the majority of analyzed products contained low CBD levels and a CBD: CBDA ratio of CBD and a CBD: CBDA ratio of >1,000 (an oil-based product). Overall, the method proved amenable to the analysis of various commercial products including oils, creams, and plant material and may be diagnostically indicative of

  2. Apparatus and method for the simultaneous detection of neutrons and ionizing electromagnetic radiation

    Science.gov (United States)

    Bell, Zane W.

    2000-01-01

    A sensor for simultaneously detecting neutrons and ionizing electromagnetic radiation comprising: a sensor for the detection of gamma radiation, the sensor defining a sensing head; the sensor further defining an output end in communication with the sensing head; and an exterior neutron-sensitive material configured to form around the sensing head; wherein the neutron-sensitive material, subsequent to the capture of the neutron, fissions into an alpha-particle and a .sup.7 Li ion that is in a first excited state in a majority of the fissions, the first excited state decaying via the emission of a single gamma ray at 478 keV which can in turn be detected by the sensing head; and wherein the sensing head can also detect the ionizing electromagnetic radiation from an incident radiation field without significant interference from the neutron-sensitive material. A method for simultaneously detecting neutrons and ionizing electromagnetic radiation comprising the steps of: providing a gamma ray sensitive detector comprising a sensing head and an output end; conforming an exterior neutron-sensitive material configured to form around the sensing head of the detector; capturing neutrons by the sensing head causing the neutron-sensitive material to fission into an alpha-particle and a .sup.7 Li ion that is in a first excited state in a majority of the fissions, the state decaying via the emission of a single gamma ray at 478 keV; sensing gamma rays entering the detector through the neutron-sensitive material; and producing an output through a readout device coupled to the output end; wherein the detector provides an output which is proportional to the energy of the absorbed ionizing electromagnetic radiation.

  3. Simultaneous Detection of Genetically Modified Organisms in a Mixture by Multiplex PCR-Chip Capillary Electrophoresis.

    Science.gov (United States)

    Patwardhan, Supriya; Dasari, Srikanth; Bhagavatula, Krishna; Mueller, Steffen; Deepak, Saligrama Adavigowda; Ghosh, Sudip; Basak, Sanjay

    2015-01-01

    An efficient PCR-based method to trace genetically modified food and feed products is in demand due to regulatory requirements and contaminant issues in India. However, post-PCR detection with conventional methods has limited sensitivity in amplicon separation that is crucial in multiplexing. The study aimed to develop a sensitive post-PCR detection method by using PCR-chip capillary electrophoresis (PCR-CCE) to detect and identify specific genetically modified organisms in their genomic DNA mixture by targeting event-specific nucleotide sequences. Using the PCR-CCE approach, novel multiplex methods were developed to detect MON531 cotton, EH 92-527-1 potato, Bt176 maize, GT73 canola, or GA21 maize simultaneously when their genomic DNAs in mixtures were amplified using their primer mixture. The repeatability RSD (RSDr) of the peak migration time was 0.06 and 3.88% for the MON531 and Bt176, respectively. The RSD (RSDR) of the Cry1Ac peak ranged from 0.12 to 0.40% in multiplex methods. The method was sensitive in resolving amplicon of size difference up to 4 bp. The PCR-CCE method is suitable to detect multiple genetically modified events in a composite DNA sample by tagging their event specific sequences.

  4. Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR

    Directory of Open Access Journals (Sweden)

    Roman Wölfel

    2012-08-01

    Full Text Available Ricin and abrin are two of the most potent plant toxins known and may be easily obtained in high yield from the seeds using rather simple technology. As a result, both toxins are potent and available toxins for criminal or terrorist acts. However, as the production of highly purified ricin or abrin requires sophisticated equipment and knowledge, it may be more likely that crude extracts would be used by non-governmental perpetrators. Remaining plant-specific nucleic acids in these extracts allow the application of a real-time PCR (qPCR assay for the detection and identification of abrin or ricin genomic material. Therefore, we have developed a duplex real-time PCR assays for simultaneous detection of ricin and abrin DNA based on the OmniMix HS bead PCR reagent mixture. Novel primers and hybridization probes were designed for detection on a SmartCycler instrument by using 5′-nuclease technology. The assay was thoroughly optimized and validated in terms of analytical sensitivity. Evaluation of the assay sensitivity by probit analysis demonstrated a 95% probability of detection at 3 genomes per reaction for ricin DNA and 1.2 genomes per reaction for abrin DNA. The suitability of the assays was exemplified by detection of ricin and abrin contaminations in a food matrix.

  5. Identification of volatiles by headspace gas chromatography with simultaneous flame ionization and mass spectrometric detection.

    Science.gov (United States)

    Tiscione, Nicholas B; Yeatman, Dustin Tate; Shan, Xiaoqin; Kahl, Joseph H

    2013-10-01

    Volatiles are frequently abused as inhalants. The methods used for identification are generally nonspecific if analyzed concurrently with ethanol or require an additional analytical procedure that employs mass spectrometry. A previously published technique utilizing a capillary flow technology splitter to simultaneously quantitate and confirm ethyl alcohol by flame ionization and mass spectrometric detection after headspace sampling and gas chromatographic separation was evaluated for the detection of inhalants. Methanol, isopropanol, acetone, acetaldehyde, toluene, methyl ethyl ketone, isoamyl alcohol, isobutyl alcohol, n-butyl alcohol, 1,1-difluoroethane, 1,1,1-trifluoroethane, 1,1,1,2-tetrafluoroethane (Norflurane, HFC-134a), chloroethane, trichlorofluoromethane (Freon®-11), dichlorodifluoromethane (Freon®-12), dichlorofluoromethane (Freon®-21), chlorodifluoromethane (Freon®-22) and 1,2-dichlorotetrafluoroethane (Freon®-114) were validated for qualitative identification by this method. The validation for qualitative identification included evaluation of matrix effects, sensitivity, carryover, specificity, repeatability and ruggedness/robustness.

  6. Simultaneous macula detection and optic disc boundary segmentation in retinal fundus images

    Science.gov (United States)

    Girard, Fantin; Kavalec, Conrad; Grenier, Sébastien; Ben Tahar, Houssem; Cheriet, Farida

    2016-03-01

    The optic disc (OD) and the macula are important structures in automatic diagnosis of most retinal diseases inducing vision defects such as glaucoma, diabetic or hypertensive retinopathy and age-related macular degeneration. We propose a new method to detect simultaneously the macula and the OD boundary. First, the color fundus images are processed to compute several maps highlighting the different anatomical structures such as vessels, the macula and the OD. Then, macula candidates and OD candidates are found simultaneously and independently using seed detectors identified on the corresponding maps. After selecting a set of macula/OD pairs, the top candidates are sent to the OD segmentation method. The segmentation method is based on local K-means applied to color coordinates in polar space followed by a polynomial fitting regularization step. Pair scores are updated, resulting in the final best macula/OD pair. The method was evaluated on two public image databases: ONHSD and MESSIDOR. The results show an overlapping area of 0.84 on ONHSD and 0.90 on MESSIDOR, which is better than recent state of the art methods. Our segmentation method is robust to contrast and illumination problems and outputs the exact boundary of the OD, not just a circular or elliptical model. The macula detection has an accuracy of 94%, which again outperforms other macula detection methods. This shows that combining the OD and macula detections improves the overall accuracy. The computation time for the whole process is 6.4 seconds, which is faster than other methods in the literature.

  7. Ethanol analysis by headspace gas chromatography with simultaneous flame-ionization and mass spectrometry detection.

    Science.gov (United States)

    Tiscione, Nicholas B; Alford, Ilene; Yeatman, Dustin Tate; Shan, Xiaoqin

    2011-09-01

    Ethanol is the most frequently identified compound in forensic toxicology. Although confirmation involving mass spectrometry is desirable, relatively few methods have been published to date. A novel technique utilizing a Dean's Switch to simultaneously quantitate and confirm ethyl alcohol by flame-ionization (FID) and mass spectrometric (MS) detection after headspace sampling and gas chromatographic separation is presented. Using 100 μL of sample, the limits of detection and quantitation were 0.005 and 0.010 g/dL, respectively. The zero-order linear range (r(2) > 0.990) was determined to span the concentrations of 0.010 to 1.000 g/dL. The coefficient of variation of replicate analyses was less than 3.1%. Quantitative accuracy was within ±8%, ±6%, ±3%, and ±1.5% at concentrations of 0.010, 0.025, 0.080, and 0.300 g/dL, respectively. In addition, 1,1-difluoroethane was validated for qualitative identification by this method. The validated FID-MS method provides a procedure for the quantitation of ethyl alcohol in blood by FID with simultaneous confirmation by MS and can also be utilized as an identification method for inhalants such as 1,1-difluoroethane.

  8. Eyewitness decisions in simultaneous and sequential lineups: a dual-process signal detection theory analysis.

    Science.gov (United States)

    Meissner, Christian A; Tredoux, Colin G; Parker, Janat F; MacLin, Otto H

    2005-07-01

    Many eyewitness researchers have argued for the application of a sequential alternative to the traditional simultaneous lineup, given its role in decreasing false identifications of innocent suspects (sequential superiority effect). However, Ebbesen and Flowe (2002) have recently noted that sequential lineups may merely bring about a shift in response criterion, having no effect on discrimination accuracy. We explored this claim, using a method that allows signal detection theory measures to be collected from eyewitnesses. In three experiments, lineup type was factorially combined with conditions expected to influence response criterion and/or discrimination accuracy. Results were consistent with signal detection theory predictions, including that of a conservative criterion shift with the sequential presentation of lineups. In a fourth experiment, we explored the phenomenological basis for the criterion shift, using the remember-know-guess procedure. In accord with previous research, the criterion shift in sequential lineups was associated with a reduction in familiarity-based responding. It is proposed that the relative similarity between lineup members may create a context in which fluency-based processing is facilitated to a greater extent when lineup members are presented simultaneously.

  9. Development and Validation of Protein Microarray Technology for Simultaneous Inflammatory Mediator Detection in Human Sera

    Directory of Open Access Journals (Sweden)

    Senthooran Selvarajah

    2014-01-01

    Full Text Available Biomarkers, including cytokines, can help in the diagnosis, prognosis, and prediction of treatment response across a wide range of disease settings. Consequently, the recent emergence of protein microarray technology, which is able to quantify a range of inflammatory mediators in a large number of samples simultaneously, has become highly desirable. However, the cost of commercial systems remains somewhat prohibitive. Here we show the development, validation, and implementation of an in-house microarray platform which enables the simultaneous quantitative analysis of multiple protein biomarkers. The accuracy and precision of the in-house microarray system were investigated according to the Food and Drug Administration (FDA guidelines for pharmacokinetic assay validation. The assay fell within these limits for all but the very low-abundant cytokines, such as interleukin- (IL- 10. Additionally, there were no significant differences between cytokine detection using our microarray system and the “gold standard” ELISA format. Crucially, future biomarker detection need not be limited to the 16 cytokines shown here but could be expanded as required. In conclusion, we detail a bespoke protein microarray system, utilizing well-validated ELISA reagents, that allows accurate, precise, and reproducible multiplexed biomarker quantification, comparable with commercial ELISA, and allowing customization beyond that of similar commercial microarrays.

  10. Advances in developing rapid, reliable and portable detection systems for alcohol.

    Science.gov (United States)

    Thungon, Phurpa Dema; Kakoti, Ankana; Ngashangva, Lightson; Goswami, Pranab

    2017-11-15

    Development of portable, reliable, sensitive, simple, and inexpensive detection system for alcohol has been an instinctive demand not only in traditional brewing, pharmaceutical, food and clinical industries but also in rapidly growing alcohol based fuel industries. Highly sensitive, selective, and reliable alcohol detections are currently amenable typically through the sophisticated instrument based analyses confined mostly to the state-of-art analytical laboratory facilities. With the growing demand of rapid and reliable alcohol detection systems, an all-round attempt has been made over the past decade encompassing various disciplines from basic and engineering sciences. Of late, the research for developing small-scale portable alcohol detection system has been accelerated with the advent of emerging miniaturization techniques, advanced materials and sensing platforms such as lab-on-chip, lab-on-CD, lab-on-paper etc. With these new inter-disciplinary approaches along with the support from the parallel knowledge growth on rapid detection systems being pursued for various targets, the progress on translating the proof-of-concepts to commercially viable and environment friendly portable alcohol detection systems is gaining pace. Here, we summarize the progress made over the years on the alcohol detection systems, with a focus on recent advancement towards developing portable, simple and efficient alcohol sensors. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Simultaneous detection of mRNA and protein stem cell markers in live cells

    Directory of Open Access Journals (Sweden)

    Bao Gang

    2009-04-01

    Full Text Available Abstract Background Biological studies and medical application of stem cells often require the isolation of stem cells from a mixed cell population, including the detection of cancer stem cells in tumor tissue, and isolation of induced pluripotent stem cells after eliciting the expression of specific genes in adult cells. Here we report the detection of Oct-4 mRNA and SSEA-1 protein in live carcinoma stem cells using respectively molecular beacon and dye-labeled antibody, aiming to establish a new method for stem cells detection and isolation. Results Quantification of Oct-4 mRNA and protein in P19 mouse carcinoma stem cells using respectively RT-PCR and immunocytochemistry confirmed that their levels drastically decreased after differentiation. To visualize Oct-4 mRNA in live stem cells, molecular beacons were designed, synthesized and validated, and the detection specificity was confirmed using control studies. We found that the fluorescence signal from Oct-4-targeting molecular beacons provides a clear discrimination between undifferentiated and retinoic acid-induced differentiated cells. Using deconvolution fluorescence microscopy, Oct-4 mRNAs were found to reside on one side of the cytosol. We demonstrated that, using a combination of Oct-4 mRNA-targeting molecular beacon with SSEA-1 antibody in flow cytometric analysis, undifferentiated stem cells can be clearly distinguished from differentiated cells. We revealed that Oct-4 targeting molecular beacons do not seem to affect stem cell biology. Conclusion Molecular beacons have the potential to provide a powerful tool for highly specific detection and isolation of stem cells, including cancer stem cells and induced pluripotent stem (iPS cells without disturbing cell physiology. It is advantageous to perform simultaneous detection of intracellular (mRNA and cell-surface (protein stem cell markers in flow cytometric analysis, which may lead to high detection sensitivity and efficiency.

  12. SIMULTANEOUS MULTI-BAND DETECTION OF LOW SURFACE BRIGHTNESS GALAXIES WITH MARKOVIAN MODELING

    Energy Technology Data Exchange (ETDEWEB)

    Vollmer, B.; Bonnarel, F.; Louys, M. [CDS, Observatoire Astronomique, UMR 7550, 11 rue de l' universite, F-67000 Strasbourg (France); Perret, B.; Petremand, M.; Lavigne, F.; Collet, Ch. [LSIIT, Universite de Strasbourg, 7, Rue Rene Descartes, F-67084 Strasbourg (France); Van Driel, W. [GEPI, Observatoire de Paris, CNRS, Universite Paris Diderot, 5 place Jules Janssen, F-92190 Meudon (France); Sabatini, S. [INAF/IASF-Roma, via Fosso de Cavaliere 100, I-00133 Roma (Italy); MacArthur, L. A., E-mail: Bernd.Vollmer@astro.unistra.fr [Herzberg Institute of Astrophysics, National Research Council of Canada, Victoria, BC V9E 2E7 (Canada)

    2013-02-01

    We present to the astronomical community an algorithm for the detection of low surface brightness (LSB) galaxies in images, called MARSIAA (MARkovian Software for Image Analysis in Astronomy), which is based on multi-scale Markovian modeling. MARSIAA can be applied simultaneously to different bands. It segments an image into a user-defined number of classes, according to their surface brightness and surroundings-typically, one or two classes contain the LSB structures. We have developed an algorithm, called DetectLSB, which allows the efficient identification of LSB galaxies from among the candidate sources selected by MARSIAA. The application of the method to two and three bands simultaneously was tested on simulated images. Based on our tests, we are confident that we can detect LSB galaxies down to a central surface brightness level of only 1.5 times the standard deviation from the mean pixel value in the image background. To assess the robustness of our method, the method was applied to a set of 18 B- and I-band images (covering 1.3 deg{sup 2} in total) of the Virgo Cluster to which Sabatini et al. previously applied a matched-filter dwarf LSB galaxy search algorithm. We have detected all 20 objects from the Sabatini et al. catalog which we could classify by eye as bona fide LSB galaxies. Our method has also detected four additional Virgo Cluster LSB galaxy candidates undetected by Sabatini et al. To further assess the completeness of the results of our method, both MARSIAA, SExtractor, and DetectLSB were applied to search for (1) mock Virgo LSB galaxies inserted into a set of deep Next Generation Virgo Survey (NGVS) gri-band subimages and (2) Virgo LSB galaxies identified by eye in a full set of NGVS square degree gri images. MARSIAA/DetectLSB recovered {approx}20% more mock LSB galaxies and {approx}40% more LSB galaxies identified by eye than SExtractor/DetectLSB. With a 90% fraction of false positives from an entirely unsupervised pipeline, a completeness of

  13. Multifunctional nanopipette for simultaneous ionic current and potential detection of nanoparticles

    Science.gov (United States)

    Panday, Namuna; He, Jin

    Nanopipette has been demonstrated as a nanopore type biosensor for DNA, protein, nanoparticle and virus analysis. In the last two decades, nanopore based technologies have made remarkable progress for single entity detection and analysis. Multifunctional nanopipette for multi-parameter detection is a new trend for nanopore based technique. We have developed a technique to fabricate multifunctional nanopipette which contains both nanopore and carbon nanoelectrode (CNE) at the nanopipette tip. It can be quickly, cheaply and reproducibly fabricated from theta pipettes. We have been able to use this multifunctional nanopieptte for simultaneous detection of ionic current and local electrical potential changes during translocation of charged gold nanoparticles (GNPs) which is used as a model experiment. The CNE functions as a local potential probe. We have demonstrated that it can detect the local potential change during translocation of a single GNP as well as collective potential change due to cluster of GNPs outside the nanopore entrance. From the potential change, we can also have insight of motion of GNPs before entering the nanopore. We have also tested insulating and biological NPs with various size and charge. Observed results have shown correlations between ionic current and potential change during translocation of these NPs. Florida International University.

  14. Methods for simultaneous detection of the cyanotoxins BMAA, DABA, and anatoxin-a in environmental samples.

    Science.gov (United States)

    Al-Sammak, Maitham Ahmed; Hoagland, Kyle D; Snow, Daniel D; Cassada, David

    2013-12-15

    Blue-green algae, also known as cyanobacteria, can produce several different groups of toxins in the environment including hepatotoxins (microcystins), neurotoxic non-protein amino acids β-methylamino-l-alanine (BMAA), and 2,4-diaminobutyric (DABA), as well as the bicyclic amine alkaloid anatoxin-a. Few studies have addressed the methods necessary for an accurate determination of cyanotoxins in environmental samples, and none have been published that can detect these cyanotoxins together in a single sample. Cyanotoxins occur in a wide range of environmental samples including water, fish, and aquatic plant samples. Using polymeric cation exchange solid phase extraction (SPE) coupled with liquid chromatography and fluorescence detection (HPLC/FD), and liquid chromatography ion trap tandem mass spectrometry (LC/MS/MS), these compounds can for the first time be simultaneously quantified in a variety of environmental sample types. The extraction method for biological samples can distinguish bound and free cyanotoxins. Detection limits for water ranged from 5 to 7 μg/L using HPLC/FD, while detection limits for and LC/MS were in the range of 0.8-3.2 μg/L. Copyright © 2013 Elsevier Ltd. All rights reserved.

  15. Simultaneous separation and detection of actinides in acidic solutions using an extractive scintillating resin.

    Science.gov (United States)

    Roane, J E; DeVol, T A

    2002-11-01

    An extractive scintillating resin was evaluated for the simultaneous separation and detection of actinides in acidic solutions. The transuranic extractive scintillating (TRU-ES) resin is composed of an inert macroporous polystyrene core impregnated with organic fluors (diphenyloxazole and 1,4-bis-(4-methyl-5-phenyl-2-oxazolyl)benzene) and an extractant (octyl(phenyl)-N,N-diisobutylcarbamoylmethylphosphine oxide in tributyl phosphate). The TRU-ES resin was packed into FEP Teflon tubing to produce a flow cell (0.2-mL free column volume), which is placed into a scintillation detection system to obtain pulse height spectra and time series data during loading and elution of actinides onto/from the resin. The alpha-particle absolute detection efficiencies ranged from 77% to 96.5%, depending on the alpha energy and quench. In addition to the on-line analyses, off-line analyses of the effluent can be conducted using conventional detection methods. The TRU-ES resin was applied to the quantification of a mixed radionuclide solution and two actual waste samples. The on-line characterization of the mixed radionuclide solution was within 10% of the reported activities whereas the agreement with the waste samples was not as good due to sorption onto the sample container walls and the oxidation state of plutonium. Agreement between the on-line and off-line analyses was within 35% of one another for both waste samples.

  16. Simultaneous electrochemical detection of dopamine and uric acid over ceria supported three dimensional gold nanoclusters

    Science.gov (United States)

    Palanisamy, Sivakumar

    2014-12-01

    CeO2 is well known for being an active material to support the growth of Au nanoclusters (Au NCs). In this work, three dimensional (3D) Au NCs were deposited on three different shaped CeO2 nanostructures such as nanoparticles (NPs), nanorod arrays (NRAs) and nanoflowers (NFs) modified Ti substrate for electrochemical simultaneous detection of dopamine (DA) and uric acid (UA). The electrodeposition of 3D Au NCs were carried out via cyclic voltammetric (CV) method at over-potential, while CeO2 nanostructures were deposited by galvanostatic constant current method under the optimized conditions. The morphology and elemental composition analysis of 3D Au NCs with CeO2 nanostructures were characterized by SEM, XRD, XPS and EDAX measurements. The electrocatalytic activity of 3D Au NCs on different CeO2 supports were thoroughly investigated by using voltammetric and amperometric techniques. According to the obtained results, CeO2 NPs supported 3D Au NCs (3D Au NCs@CeO2 NPs) displayed strong signal for DA as compared to that of CeO2 NRAs (3D Au NCs@CeO2 NRAs) and CeO2 NFs supported 3D Au NCs (3D Au NCs@CeO2 NFs). In addition, the 3D Au NCs@CeO2 NPs electrode resulted in more sensitive and simultaneous detection of DA in the presence of excess UA. Thus, the 3D Au NCs@CeO2 NPs electrode can practically be applied for the detection of DA using biological samples.

  17. Simultaneous detection of IgG antibodies associated with viral hemorrhagic fever by a multiplexed Luminex-based immunoassay.

    Science.gov (United States)

    Wu, Wei; Zhang, Shuo; Qu, Jing; Zhang, Quanfu; Li, Chuan; Li, Jiandong; Jin, Cong; Liang, Mifang; Li, Dexin

    2014-07-17

    Viral hemorrhagic fevers (VHFs) are worldwide diseases caused by several kinds of viruses. With the emergence of new viruses, advanced diagnostic methods are urgently needed for identification of VHFs. Based on Luminex xMAP technology, a rapid, sensitive, multi-pathogen and high-throughput method which could simultaneously detect hemorrhagic fever viruses (HFVs) specific IgG antibodies was developed. Recombinant antigens of nine HFVs including Hantaan virus (HTNV), Seoul virus (SEOV), Puumala virus (PUUV), Andes virus (ANDV), Sin Nombre virus (SNV), Crimean-Congo hemorrhagic fever virus (CCHFV), Rift Valley fever virus (RVFV), Severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) and dengue virus (DENV) were produced and purified from a prokaryotic expression system and the influence of the coupling amount was investigated. Cross-reactions among antigens and their rabbit immune sera were evaluated. Serum samples collected from 51 laboratory confirmed hemorrhagic fever with renal syndrome (HFRS) patients, 43 confirmed SFTS patients and 88 healthy donors were analyzed. Results showed that recombinant nucleocapsid protein of the five viruses belonging to the genus Hantavirus, had serological cross-reactivity with their corresponding rabbit immune sera, but not apparent with immune sera of other four viruses. Evaluation of this new method with clinical serum samples showed 98.04% diagnostic sensitivity for HFRS, 90.70% for SFTS detection and the specificity was ranging from 66.67% to 100.00%. The multiplexed Luminex-based immunoassay has firstly been established in our study, which provides a potentially reliable diagnostic tool for IgG antibody detection of VHFs. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Reliability and minimal detectable difference in multisegment foot kinematics during shod walking and running.

    Science.gov (United States)

    Milner, Clare E; Brindle, Richard A

    2016-01-01

    There has been increased interest recently in measuring kinematics within the foot during gait. While several multisegment foot models have appeared in the literature, the Oxford foot model has been used frequently for both walking and running. Several studies have reported the reliability for the Oxford foot model, but most studies to date have reported reliability for barefoot walking. The purpose of this study was to determine between-day (intra-rater) and within-session (inter-trial) reliability of the modified Oxford foot model during shod walking and running and calculate minimum detectable difference for common variables of interest. Healthy adult male runners participated. Participants ran and walked in the gait laboratory for five trials of each. Three-dimensional gait analysis was conducted and foot and ankle joint angle time series data were calculated. Participants returned for a second gait analysis at least 5 days later. Intraclass correlation coefficients and minimum detectable difference were determined for walking and for running, to indicate both within-session and between-day reliability. Overall, relative variables were more reliable than absolute variables, and within-session reliability was greater than between-day reliability. Between-day intraclass correlation coefficients were comparable to those reported previously for adults walking barefoot. It is an extension in the use of the Oxford foot model to incorporate wearing a shoe while maintaining marker placement directly on the skin for each segment. These reliability data for walking and running will aid in the determination of meaningful differences in studies which use this model during shod gait. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. The reliability of magnetic resonance imaging in traumatic brain injury lesion detection

    NARCIS (Netherlands)

    Geurts, B.H.J.; Andriessen, T.M.J.C.; Goraj, B.M.; Vos, P.E.

    2012-01-01

    Objective: This study compares inter-rater-reliability, lesion detection and clinical relevance of T2-weighted imaging (T2WI), Fluid Attenuated Inversion Recovery (FLAIR), T2*-gradient recalled echo (T2*-GRE) and Susceptibility Weighted Imaging (SWI) in Traumatic Brain Injury (TBI). Methods: Three

  20. Electronic logic to enhance switch reliability in detecting openings and closures of redundant switches

    Science.gov (United States)

    Cooper, James A.

    1986-01-01

    A logic circuit is used to enhance redundant switch reliability. Two or more switches are monitored for logical high or low output. The output for the logic circuit produces a redundant and failsafe representation of the switch outputs. When both switch outputs are high, the output is high. Similarly, when both switch outputs are low, the logic circuit's output is low. When the output states of the two switches do not agree, the circuit resolves the conflict by memorizing the last output state which both switches were simultaneously in and produces the logical complement of this output state. Thus, the logic circuit of the present invention allows the redundant switches to be treated as if they were in parallel when the switches are open and as if they were in series when the switches are closed. A failsafe system having maximum reliability is thereby produced.

  1. Simultaneous determination of eight cyclopolypeptide antibiotics in feed by high performance liquid chromatography coupled with evaporation light scattering detection.

    Science.gov (United States)

    Song, Xuqin; Xie, Jingmeng; Zhang, Meiyu; Zhang, Yingxia; Li, Jiufeng; Huang, Qiwen; He, Limin

    2018-02-15

    A high throughput, reliable and reproducible analysis strategy based on high performance liquid chromatography combined to evaporative light scattering detector (HPLC-ELSD) was developed for simultaneous determination of eight cyclopolypeptide antibiotics including vancomycin, polymyxin B (polymyxin B1 and polymyxin B2), polymyxin E (colistin A and colistin B), teicoplanin, bacitracin A, daptomycin and virginiamycin M1 in animal Feed. Feed samples were extracted with methanol-2% formic acid aqueous solution, followed by a solid-phase extraction step using a HLB cartridge. Under the optimum chromatographic conditions and ELSD parameters, target compounds were separated well on a short column filled with biphenyl stationary phase. The method was developed in accordance with pig complete feed and then extended to detect polypeptide antibiotics in piglet premix, pig feed additive, poultry complete feed and fattening pig premix. The results showed that logarithmic calibration curves of eight analytes were linear (r 2  > 0.99) within the concentration range of 5-200 mg mL -1 . The developed method provided good accuracy and precision for quantification of eight polypeptides in five kinds of feeds with recoveries ranging from 72.0% to 105.4% with relative standard deviations antibiotics in commercial feed. Copyright © 2018 Elsevier B.V. All rights reserved.

  2. Simultaneous determination of quinolones for veterinary use by high-performance liquid chromatography with electrochemical detection.

    Science.gov (United States)

    Rodríguez Cáceres, M I; Guiberteau Cabanillas, A; Galeano Díaz, T; Martínez Cañas, M A

    2010-02-01

    A selective method based on high-performance liquid chromatography with electrochemical detection (HPLC-ECD) has been developed to enable simultaneous determination of three fluoroquinolones (FQs), namely danofloxacin (DANO), difloxacin (DIFLO) and sarafloxacin (SARA). The fluoroquinolones are separated on a Novapack C-18 column and detected in a high sensitivity amperometric cell at a potential of +0.8 V. Solid-phase extraction was used for the extraction of the analytes in real samples. The range of concentration examined varied from 10 to 150 ng g(-1) for danofloxacin, from 25 to 100 ng g(-1) for sarafloxacin and from 50 to 315 ng g(-1) for difloxacin, respectively. The method presents detection limits under 10 ng g(-1) and recoveries around 90% for the three analytes have been obtained in the experiments with fortified samples. This HPLC-ECD approach can be useful in the routine analysis of antibacterial residues being less expensive and less complicated than other more powerful tools as hyphenated techniques. 2009 Elsevier B.V. All rights reserved.

  3. Quantitative 3D-KPFM imaging with simultaneous electrostatic force and force gradient detection

    International Nuclear Information System (INIS)

    Collins, L; Rodriguez, B J; Okatan, M B; Li, Q; Kravenchenko, I I; Lavrik, N V; Kalinin, S V; Jesse, S

    2015-01-01

    Kelvin probe force microscopy (KPFM) is a powerful characterization technique for imaging local electrochemical and electrostatic potential distributions and has been applied across a broad range of materials and devices. Proper interpretation of the local KPFM data can be complicated, however, by convolution of the true surface potential under the tip with additional contributions due to long range capacitive coupling between the probe (e.g. cantilever, cone, tip apex) and the sample under test. In this work, band excitation (BE)-KPFM is used to negate such effects. In contrast to traditional single frequency KPFM, multifrequency BE-KPFM is shown to afford dual sensitivity to both the electrostatic force and the force gradient detection, analogous to simultaneous amplitude modulated and frequency modulated KPFM imaging. BE-KPFM is demonstrated on a Pt/Au/SiO x test structure and electrostatic force gradient detection is found to lead to an improved lateral resolution compared to electrostatic force detection. Finally, a 3D-KPFM imaging technique is developed. Force volume (FV) BE-KPFM allows the tip–sample distance dependence of the electrostatic interactions (force and force gradient) to be recorded at each point across the sample surface. As such, FVBE-KPFM provides a much needed pathway towards complete tip–sample capacitive de-convolution in KPFM measurements and will enable quantitative surface potential measurements with nanoscale resolution. (paper)

  4. Aptamer/quantum dot-based simultaneous electrochemical detection of multiple small molecules

    Energy Technology Data Exchange (ETDEWEB)

    Zhang Haixia [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Jiang Bingying [School of Chemistry and Chemical Engineering, Chongqing University of Technology, Chongqing 400040 (China); Xiang Yun, E-mail: yunatswu@swu.edu.cn [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Zhang Yuyong; Chai Yaqin [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Yuan Ruo, E-mail: yuanruo@swu.edu.cn [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China)

    2011-03-04

    A novel strategy for 'signal on' and sensitive one-spot simultaneous detection of multiple small molecular analytes based on electrochemically encoded barcode quantum dot (QD) tags is described. The target analytes, adenosine triphosphate (ATP) and cocaine, respectively, are sandwiched between the corresponding set of surface-immobilized primary binding aptamers and the secondary binding aptamer/QD bioconjugates. The captured QDs yield distinct electrochemical signatures after acid dissolution, whose position and size reflect the identity and level, respectively, of the corresponding target analytes. Due to the inherent amplification feature of the QD labels and the 'signal on' detection scheme, as well as the sensitive monitoring of the metal ions released upon acid dissolution of the QD labels, low detection limits of 30 nM and 50 nM were obtained for ATP and cocaine, respectively, in our assays. Our multi-analyte sensing system also shows high specificity to target analytes and promising applicability to complex sample matrix, which makes the proposed assay protocol an attractive route for screening of small molecules in clinical diagnosis.

  5. Simultaneous Detection of 13 Key Bacterial Respiratory Pathogens by Combination of Multiplex PCR and Capillary Electrophoresis.

    Science.gov (United States)

    Jiang, Lu Xi; Ren, Hong Yu; Zhou, Hai Jian; Zhao, Si Hong; Hou, Bo Yan; Yan, Jian Ping; Qin, Tian; Chen, Yu

    2017-08-01

    Lower respiratory tract infections continue to pose a significant threat to human health. It is important to accurately and rapidly detect respiratory bacteria. To compensate for the limits of current respiratory bacteria detection methods, we developed a combination of multiplex polymerase chain reaction (PCR) and capillary electrophoresis (MPCE) assay to detect thirteen bacterial pathogens responsible for lower respiratory tract infections, including Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Mycoplasma pneumoniae, Legionella spp., Bordetella pertussis, Mycobacterium tuberculosis complex, Corynebacterium diphtheriae, and Streptococcus pyogenes. Three multiplex PCR reactions were built, and the products were analyzed by capillary electrophoresis using the high-throughput DNA analyzer. The specificity of the MPCE assay was examined and the detection limit was evaluated using DNA samples from each bacterial strain and the simulative samples of each strain. This assay was further evaluated using 152 clinical specimens and compared with real-time PCR reactions. For this assay, three nested-multiplex-PCRs were used to detect these clinical specimens. The detection limits of the MPCE assay for the 13 pathogens were very low and ranged from 10-7 to 10-2 ng/μL. Furthermore, analysis of the 152 clinical specimens yielded a specificity ranging from 96.5%-100.0%, and a sensitivity of 100.0% for the 13 pathogens. This study revealed that the MPCE assay is a rapid, reliable, and high-throughput method with high specificity and sensitivity. This assay has great potential in the molecular epidemiological survey of respiratory pathogens. Copyright © 2017 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

  6. Highly multiplexed simultaneous detection of RNAs and proteins in single cells.

    Science.gov (United States)

    Frei, Andreas P; Bava, Felice-Alessio; Zunder, Eli R; Hsieh, Elena W Y; Chen, Shih-Yu; Nolan, Garry P; Gherardini, Pier Federico

    2016-03-01

    To enable the detection of expression signatures specific to individual cells, we developed PLAYR (proximity ligation assay for RNA), a method for highly multiplexed transcript quantification by flow and mass cytometry that is compatible with standard antibody staining. When used with mass cytometry, PLAYR allowed for the simultaneous quantification of more than 40 different mRNAs and proteins. In primary cells, we quantified multiple transcripts, with the identity and functional state of each analyzed cell defined on the basis of the expression of a separate set of transcripts or proteins. By expanding high-throughput deep phenotyping of cells beyond protein epitopes to include RNA expression, PLAYR opens a new avenue for the characterization of cellular metabolism.

  7. Simultaneous Fault Detection and Sensor Selection for Condition Monitoring of Wind Turbines

    Directory of Open Access Journals (Sweden)

    Wenna Zhang

    2016-04-01

    Full Text Available Data collected from the supervisory control and data acquisition (SCADA system are used widely in wind farms to obtain operation and performance information about wind turbines. The paper presents a three-way model by means of parallel factor analysis (PARAFAC for wind turbine fault detection and sensor selection, and evaluates the method with SCADA data obtained from an operational farm. The main characteristic of this new approach is that it can be used to simultaneously explore measurement sample profiles and sensors profiles to avoid discarding potentially relevant information for feature extraction. With K-means clustering method, the measurement data indicating normal, fault and alarm conditions of the wind turbines can be identified, and the sensor array can be optimised for effective condition monitoring.

  8. Etched FBG coated with polyimide for simultaneous detection the salinity and temperature

    Science.gov (United States)

    Luo, Dong; Ma, Jianxun; Ibrahim, Zainah; Ismail, Zubaidah

    2017-06-01

    In marine environment, concrete structures can corrode because of the PH alkalinity of concrete paste; and the salinity PH is heavily related with the concentration of salt in aqueous solutions. In this study, an optical fiber salinity sensor is proposed on the basis of an etched FBG (EFBG) coated with a layer of polyimide. Chemical etching is employed to reduce the diameter of FBG and to excite Cladding Mode Resonance Wavelengths (CMRWs). CMRW and Fundamental Mode Resonance Wavelength (FMRW) can be used to measure the Refractive index (RI) and temperature of salinity. The proposed sensor is then characterized with a matrix equation. Experimental results show that FMRW and 5th CMRW have the detection sensitivities of 15.407 and 125.92 nm/RIU for RI and 0.0312 and 0.0435 nm/°C for temperature, respectively. The proposed sensor can measure salinity and temperature simultaneously.

  9. Simultaneous Detection of Bovine Theileria and Babesia Species by Reverse Line Blot Hybridization

    Science.gov (United States)

    Gubbels, J. M.; de Vos, A. P.; van der Weide, M.; Viseras, J.; Schouls, L. M.; de Vries, E.; Jongejan, F.

    1999-01-01

    A reverse line blot (RLB) assay was developed for the identification of cattle carrying different species of Theileria and Babesia simultaneously. We included Theileria annulata, T. parva, T. mutans, T. taurotragi, and T. velifera in the assay, as well as parasites belonging to the T. sergenti-T. buffeli-T. orientalis group. The Babesia species included were Babesia bovis, B. bigemina, and B. divergens. The assay employs one set of primers for specific amplification of the rRNA gene V4 hypervariable regions of all Theileria and Babesia species. PCR products obtained from blood samples were hybridized to a membrane onto which nine species-specific oligonucleotides were covalently linked. Cross-reactions were not observed between any of the tested species. No DNA sequences from Bos taurus or other hemoparasites (Trypanosoma species, Cowdria ruminantium, Anaplasma marginale, and Ehrlichia species) were amplified. The sensitivity of the assay was determined at 0.000001% parasitemia, enabling detection of the carrier state of most parasites. Mixed DNAs from five different parasites were correctly identified. Moreover, blood samples from cattle experimentally infected with two different parasites reacted only with the corresponding species-specific oligonucleotides. Finally, RLB was used to screen blood samples collected from carrier cattle in two regions of Spain. T. annulata, T. orientalis, and B. bigemina were identified in these samples. In conclusion, the RLB is a versatile technique for simultaneous detection of all bovine tick-borne protozoan parasites. We recommend its use for integrated epidemiological monitoring of tick-borne disease, since RLB can also be used for screening ticks and can easily be expanded to include additional hemoparasite species. PMID:10325324

  10. Reliability of recordings of subgingival calculus detected using an ultrasonic device.

    Science.gov (United States)

    Corraini, Priscila; López, Rodrigo

    2015-04-01

    To assess the intra-examiner reliability of recordings of subgingival calculus detected using an ultrasonic device, and to investigate the influence of subject-, tooth- and site-level factors on the reliability of these subgingival calculus recordings. On two occasions, within a 1-week interval, 147 adult periodontitis patients received a full-mouth clinical periodontal examination by a single trained examiner. Duplicate subgingival calculus recordings, in six sites per tooth, were obtained using an ultrasonic device for calculus detection and removal. Agreement was observed in 65 % of the 22,584 duplicate subgingival calculus recordings, ranging 45 % to 83 % according to subject. Using hierarchical modeling, disagreements in the subgingival calculus duplicate recordings were more likely in all other sites than the mid-buccal, and in sites harboring supragingival calculus. Disagreements were less likely in sites with PD ≥  4 mm and with furcation involvement  ≥  degree 2. Bleeding on probing or suppuration did not influence the reliability of subgingival calculus. At the subject-level, disagreements were less likely in patients presenting with the highest and lowest extent categories of the covariate subgingival calculus. The reliability of subgingival calculus recordings using the ultrasound technology is reasonable. The results of the present study suggest that the reliability of subgingival calculus recordings is not influenced by the presence of inflammation. Moreover, subgingival calculus can be more reliably detected using the ultrasound device at sites with higher need for periodontal therapy, i.e., sites presenting with deep pockets and premolars and molars with furcation involvement.

  11. Test-retest reliability of myofascial trigger point detection in hip and thigh areas.

    Science.gov (United States)

    Rozenfeld, E; Finestone, A S; Moran, U; Damri, E; Kalichman, L

    2017-10-01

    Myofascial trigger points (MTrP's) are a primary source of pain in patients with musculoskeletal disorders. Nevertheless, they are frequently underdiagnosed. Reliable MTrP palpation is the necessary for their diagnosis and treatment. The few studies that have looked for intra-tester reliability of MTrPs detection in upper body, provide preliminary evidence that MTrP palpation is reliable. Reliability tests for MTrP palpation on the lower limb have not yet been performed. To evaluate inter- and intra-tester reliability of MTrP recognition in hip and thigh muscles. Reliability study. 21 patients (15 males and 6 females, mean age 21.1 years) referred to the physical therapy clinic, 10 with knee or hip pain and 11 with pain in an upper limb, low back, shin or ankle. Two experienced physical therapists performed the examinations, blinded to the subjects' identity, medical condition and results of the previous MTrP evaluation. Each subject was evaluated four times, twice by each examiner in a random order. Dichotomous findings included a palpable taut band, tenderness, referred pain, and relevance of referred pain to patient's complaint. Based on these, diagnosis of latent MTrP's or active MTrP's was established. The evaluation was performed on both legs and included a total of 16 locations in the following muscles: rectus femoris (proximal), vastus medialis (middle and distal), vastus lateralis (middle and distal) and gluteus medius (anterior, posterior and distal). Inter- and intra-tester reliability (Cohen's kappa (κ)) values for single sites ranged from -0.25 to 0.77. Median intra-tester reliability was 0.45 and 0.46 for latent and active MTrP's, and median inter-tester reliability was 0.51 and 0.64 for latent and active MTrPs, respectively. The examination of the distal vastus medialis was most reliable for latent and active MTrP's (intra-tester k = 0.27-0.77, inter-tester k = 0.77 and intra-tester k = 0.53-0.72, inter-tester k = 0.72, correspondingly

  12. Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria.

    Science.gov (United States)

    Zhang, D F; Zhang, Q Q; Li, A H

    2014-11-01

    Species of genus Aeromonas, Vibrio, Edwardsiella and Streptococcus are the most common fish pathogenic bacteria that cause economically devastating losses in aquaculture. A multiplex polymerase chain reaction (mPCR) was developed for the simultaneous detection and differentiation of the four genera of fish pathogenic bacteria. Through the use of genus-specific primers instead of species-specific ones, the current mPCR covered much more target bacterial species compared with previously reported species-specific mPCR methods. The specificity of the four putative genus-specific primers was validated experimentally while used exclusively (uniplex PCR) or combined (mPCR) against bacterial genomic DNA templates of the target bacteria and nontarget bacteria. The PCR amplicons for the following genera were obtained as expected: Aeromonas (875 bp), Vibrio (524 bp), Edwardsiella (302 bp) and Streptococcus (197 bp), and the fragments could be separated clearly on the agarose gel electrophoresis. The mPCR did not produce nonspecific amplification products when used to amplify 21 nontarget species of bacteria. The mPCR detection limits for each target bacterial genera were 50 colony-forming units (CFU) in pure culture and 100 CFU in fish tissue samples. In conclusion, the mPCR assay was proven to be a powerful alternative to the conventional culture-based method, given its rapid, specific, sensitive and reliable detection of target pathogens. The fish pathogenic bacteria of genus Aeromonas, Vibrio, Edwardsiella and Streptococcus frequently cause severe outbreaks of diseases in cultured fish, and the genus-specific multiplex PCR assay developed in this study can detect the bacteria of the four genera when present in the samples either alone or mixed. The mPCR assay is expected to identify the causative agents more efficiently than uniplex PCR or species-specific multiplex PCR for clinical diagnosis, resulting in the earlier implementation of control measures. This m

  13. Development of a GeXP-multiplex PCR assay for the simultaneous detection and differentiation of six cattle viruses.

    Directory of Open Access Journals (Sweden)

    Qing Fan

    Full Text Available Foot-and-mouth disease virus (FMDV, Bluetongue virus (BTV, Vesicular stomatitis Virus (VSV, Bovine viral diarrheal (BVDV, Bovine rotavirus (BRV, and Bovine herpesvirus 1 (IBRV are common cattle infectious viruses that cause a great economic loss every year in many parts of the world. A rapid and high-throughput GenomeLab Gene Expression Profiler (GeXP analyzer-based multiplex PCR assay was developed for the simultaneous detection and differentiation of these six cattle viruses. Six pairs of chimeric primers consisting of both the gene-specific primer and a universal primer were designed and used for amplification. Then capillary electrophoresis was used to separate the fluorescent labeled PCR products according to the amplicons size. The specificity of GeXP-multiplex PCR assay was examined with samples of the single template and mixed template of six viruses. The sensitivity was evaluated using the GeXP-multiplex PCR assay on serial 10-fold dilutions of ssRNAs obtained via in vitro transcription. To further evaluate the reliability, 305 clinical samples were tested by the GeXP-multiplex PCR assay. The results showed that the corresponding virus specific fragments of genes were amplified. The detection limit of the GeXP-multiplex PCR assay was 100 copies/μL in a mixed sample of ssRNAs containing target genes of six different cattle viruses, whereas the detection limit for the Gexp-mono PCR assay for a single target gene was 10 copies/μL. In detection of viruses in 305 clinical samples, the results of GeXP were consistent with simplex real-time PCR. Analysis of positive samples by sequencing demonstrated that the GeXP-multiplex PCR assay had no false positive samples of nonspecific amplification. In conclusion, this GeXP-multiplex PCR assay is a high throughput, specific, sensitive, rapid and simple method for the detection and differentiation of six cattle viruses. It is an effective tool that can be applied for the rapid differential diagnosis

  14. Rapid focused sequencing: a multiplexed assay for simultaneous detection and strain typing of Bacillus anthracis, Francisella tularensis, and Yersinia pestis.

    Directory of Open Access Journals (Sweden)

    Rosemary S Turingan

    Full Text Available BACKGROUND: The intentional release of Bacillus anthracis in the United States in 2001 has heightened concern about the use of pathogenic microorganisms in bioterrorism attacks. Many of the deadliest bacteria, including the Class A Select Agents Bacillus anthracis, Francisella tularensis, and Yersinia pestis, are highly infectious via the pulmonary route when released in aerosolized form. Hence, rapid, sensitive, and reliable methods for detection of these biothreats and characterization of their potential impact on the exposed population are of critical importance to initiate and support rapid military, public health, and clinical responses. METHODOLOGY/PRINCIPAL FINDINGS: We have developed microfluidic multiplexed PCR and sequencing assays based on the simultaneous interrogation of three pathogens per assay and ten loci per pathogen. Microfluidic separation of amplified fluorescently labeled fragments generated characteristic electrophoretic signatures for identification of each agent. The three sets of primers allowed significant strain typing and discrimination from non-pathogenic closely-related species and environmental background strains based on amplicon sizes alone. Furthermore, sequencing of the 10 amplicons per pathogen, termed "Rapid Focused Sequencing," allowed an even greater degree of strain discrimination and, in some cases, can be used to determine virulence. Both amplification and sequencing assays were performed in microfluidic biochips developed for fast thermal cycling and requiring 7 µL per reaction. The 30-plex sequencing assay resulted in genotypic resolution of 84 representative strains belonging to each of the three biothreat species. CONCLUSIONS/SIGNIFICANCE: The microfluidic multiplexed assays allowed identification and strain differentiation of the biothreat agents Bacillus anthracis, Francisella tularensis, and Yersinia pestis and clear discrimination from closely-related species and several environmental

  15. Capillary gel electrophoresis-coupled aptamer enzymatic cleavage protection strategy for the simultaneous detection of multiple small analytes.

    Science.gov (United States)

    Perrier, Sandrine; Zhu, Zhenyu; Fiore, Emmanuelle; Ravelet, Corinne; Guieu, Valérie; Peyrin, Eric

    2014-05-06

    This novel, multi small-analyte sensing strategy is the result of combining the target-induced aptamer enzymatic protection approach with the CGE-LIF (capillary gel electrophoresis with laser-induced fluorescence) technique. The implemented assay principle is based on an analysis of the phosphodiesterase I (PDE I)-mediated size variation of a fluorescein-labeled aptamer (FApt), the enzyme catalyzing the removal of nucleotides from DNA in the 3' to 5' direction. In the absence of the target, the unfolded aptamer was enzymatically cleaved into short DNA fragments. Upon target binding, the DNA substrate was partially protected against enzymatic hydrolysis. The amount of bound aptamer remaining after the exonuclease reaction was proportional to the concentration of the target. The CGE technique, which was used to determine the separation of FApt species from DNA digested products, permitted the quantification of adenosine (A), ochratoxin A (O), and tyrosinamide (T) under the same optimized enzymatic conditions. This assay strategy was subsequently applied to the simultaneous detection of A, O, and T in a single capillary under buffered conditions using corresponding FApt probes of different lengths (23, 36, and 49 nucleotides, respectively). Additionally, the detection of these three small molecules was successfully achieved in a complex medium (diluted, heat-treated human serum) showing a good recovery. It is worth noting that the multiplexed analysis was accomplished for targets with different charge states by using aptamers possessing various structural features. This sensing platform constitutes a rationalized and reliable approach with an expanded potential for a high-throughput determination of small analytes in a single capillary.

  16. Matrix approach to the simultaneous detection of multiple potato pathogens by real-time PCR.

    Science.gov (United States)

    Nikitin, M M; Statsyuk, N V; Frantsuzov, P A; Dzhavakhiya, V G; Golikov, A G

    2018-03-01

    Create a method for highly sensitive, selective, rapid and easy-to-use detection and identification of economically significant potato pathogens, including viruses, bacteria and oomycetes, be it single pathogen, or a range of various pathogens occurring simultaneously. Test-systems for real-time PCR, operating in the unified amplification regime, have been developed for Phytophthora infestans, Pectobacterium atrosepticum, Dickeya dianthicola, Dickeya solani, Ralstonia solanacearum, Pectobacterium carotovorum, Clavibacter michiganensis subsp. sepedonicus, potato viruses Y (ordinary and necrotic forms as well as indiscriminative test system, detecting all forms), A, X, S, M, potato leaf roll virus, potato mop top virus and potato spindle tuber viroid. The test-systems (including polymerase and revertase) were immobilized and lyophilized in miniature microreactors (1·2 μl) on silicon DNA/RNA microarrays (micromatrices) to be used with a mobile AriaDNA ® amplifier. Preloaded 30-reaction micromatrices having shelf life of 3 and 6 months (for RNA- and DNA-based pathogens, respectively) at room temperature with no special conditions were successfully tested on both reference and field samples in comparison with traditional ELISA and microbiological methods, showing perfect performance and sensitivity (1 pg). The accurate, rapid and user-friendly diagnostic system in a micromatrix format may significantly contribute to pathogen screening and phytopathological studies. © 2018 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of The Society for Applied Microbiology.

  17. Biomarker Detection in Association Studies: Modeling SNPs Simultaneously via Logistic ANOVA

    KAUST Repository

    Jung, Yoonsuh; Huang, Jianhua Z.; Hu, Jianhua

    2014-01-01

    In genome-wide association studies, the primary task is to detect biomarkers in the form of Single Nucleotide Polymorphisms (SNPs) that have nontrivial associations with a disease phenotype and some other important clinical/environmental factors. However, the extremely large number of SNPs comparing to the sample size inhibits application of classical methods such as the multiple logistic regression. Currently the most commonly used approach is still to analyze one SNP at a time. In this paper, we propose to consider the genotypes of the SNPs simultaneously via a logistic analysis of variance (ANOVA) model, which expresses the logit transformed mean of SNP genotypes as the summation of the SNP effects, effects of the disease phenotype and/or other clinical variables, and the interaction effects. We use a reduced-rank representation of the interaction-effect matrix for dimensionality reduction, and employ the L 1-penalty in a penalized likelihood framework to filter out the SNPs that have no associations. We develop a Majorization-Minimization algorithm for computational implementation. In addition, we propose a modified BIC criterion to select the penalty parameters and determine the rank number. The proposed method is applied to a Multiple Sclerosis data set and simulated data sets and shows promise in biomarker detection.

  18. A bio-image sensor for simultaneous detection of multi-neurotransmitters.

    Science.gov (United States)

    Lee, You-Na; Okumura, Koichi; Horio, Tomoko; Iwata, Tatsuya; Takahashi, Kazuhiro; Hattori, Toshiaki; Sawada, Kazuaki

    2018-03-01

    We report here a new bio-image sensor for simultaneous detection of spatial and temporal distribution of multi-neurotransmitters. It consists of multiple enzyme-immobilized membranes on a 128 × 128 pixel array with read-out circuit. Apyrase and acetylcholinesterase (AChE), as selective elements, are used to recognize adenosine 5'-triphosphate (ATP) and acetylcholine (ACh), respectively. To enhance the spatial resolution, hydrogen ion (H + ) diffusion barrier layers are deposited on top of the bio-image sensor and demonstrated their prevention capability. The results are used to design the space among enzyme-immobilized pixels and the null H + sensor to minimize the undesired signal overlap by H + diffusion. Using this bio-image sensor, we can obtain H + diffusion-independent imaging of concentration gradients of ATP and ACh in real-time. The sensing characteristics, such as sensitivity and detection of limit, are determined experimentally. With the proposed bio-image sensor the possibility exists for customizable monitoring of the activities of various neurochemicals by using different kinds of proton-consuming or generating enzymes. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Biomarker Detection in Association Studies: Modeling SNPs Simultaneously via Logistic ANOVA

    KAUST Repository

    Jung, Yoonsuh

    2014-10-02

    In genome-wide association studies, the primary task is to detect biomarkers in the form of Single Nucleotide Polymorphisms (SNPs) that have nontrivial associations with a disease phenotype and some other important clinical/environmental factors. However, the extremely large number of SNPs comparing to the sample size inhibits application of classical methods such as the multiple logistic regression. Currently the most commonly used approach is still to analyze one SNP at a time. In this paper, we propose to consider the genotypes of the SNPs simultaneously via a logistic analysis of variance (ANOVA) model, which expresses the logit transformed mean of SNP genotypes as the summation of the SNP effects, effects of the disease phenotype and/or other clinical variables, and the interaction effects. We use a reduced-rank representation of the interaction-effect matrix for dimensionality reduction, and employ the L 1-penalty in a penalized likelihood framework to filter out the SNPs that have no associations. We develop a Majorization-Minimization algorithm for computational implementation. In addition, we propose a modified BIC criterion to select the penalty parameters and determine the rank number. The proposed method is applied to a Multiple Sclerosis data set and simulated data sets and shows promise in biomarker detection.

  20. Improvement on simultaneous determination of chromium species in aqueous solution by ion chromatography and chemiluminescence detection

    DEFF Research Database (Denmark)

    Gammelgaard, Bente; Liao, Y.P.; Jons, O.

    1997-01-01

    A sensitive method for the simultaneous determination of chromium(III) and chromium(VI) was chromatography and chemiluminescence detection. Two Dionex ion-exchange guard columns in series, CG5 and AG7, were used to separate chromium(III) from chromium(VI). Chromium(VI) was reduced by potassium......, the stabilities of reductant and luminol solutions were studied. The linear range of the calibration curve for chromium(III) and chromium(VI) was 1-400 mu g l(-1). The detection limit was 0.12 mu g l(-1) for chromium(III) and 0.09 mu g l(-1) for chromium(VI), respectively. The precision at the 20 mu g l(-1) level...... was 1.4% for chromium(III) and 2.5% for chromium(VI), respectively. The accuracy of the chromium(III) determination was determined by analysis of the NIST standard reference material 1643c, Trace elements in water with the result 19.1 +/- 1.0 mu g Cr(III) l(-1) (certified value 19.0 +/- 0.6 mu g Cr...

  1. Reliability assessment for thickness measurements of pipe wall using probability of detection

    International Nuclear Information System (INIS)

    Nakamoto, Hiroyuki; Kojima, Fumio; Kato, Sho

    2013-01-01

    This paper proposes a reliability assessment method for thickness measurements of pipe wall using probability of detection (POD). Thicknesses of pipes are measured by qualified inspectors with ultrasonic thickness gauges. The inspection results are affected by human factors of the inspectors and include some errors, because the inspectors have different experiences and frequency of inspections. In order to ensure reliability for inspection results, first, POD evaluates experimental results of pipe-wall thickness inspection. We verify that the results have differences depending on inspectors including qualified inspectors. Second, two human factors that affect POD are indicated. Finally, it is confirmed that POD can identify the human factors and ensure reliability for pipe-wall thickness inspections. (author)

  2. Comparing apples and oranges: fold-change detection of multiple simultaneous inputs.

    Directory of Open Access Journals (Sweden)

    Yuval Hart

    Full Text Available Sensory systems often detect multiple types of inputs. For example, a receptor in a cell-signaling system often binds multiple kinds of ligands, and sensory neurons can respond to different types of stimuli. How do sensory systems compare these different kinds of signals? Here, we consider this question in a class of sensory systems - including bacterial chemotaxis- which have a property known as fold-change detection: their output dynamics, including amplitude and response time, depends only on the relative changes in signal, rather than absolute changes, over a range of several decades of signal. We analyze how fold-change detection systems respond to multiple signals, using mathematical models. Suppose that a step of fold F1 is made in input 1, together with a step of F2 in input 2. What total response does the system provide? We show that when both input signals impact the same receptor with equal number of binding sites, the integrated response is multiplicative: the response dynamics depend only on the product of the two fold changes, F1F2. When the inputs bind the same receptor with different number of sites n1 and n2, the dynamics depend on a product of power laws, [Formula: see text]. Thus, two input signals which vary over time in an inverse way can lead to no response. When the two inputs affect two different receptors, other types of integration may be found and generally the system is not constrained to respond according to the product of the fold-change of each signal. These predictions can be readily tested experimentally, by providing cells with two simultaneously varying input signals. The present study suggests how cells can compare apples and oranges, namely by comparing each to its own background level, and then multiplying these two fold-changes.

  3. Gold nanoparticles enhanced SERS aptasensor for the simultaneous detection of Salmonella typhimurium and Staphylococcus aureus.

    Science.gov (United States)

    Zhang, Hui; Ma, Xiaoyuan; Liu, Ying; Duan, Nuo; Wu, Shijia; Wang, Zhouping; Xu, Baocai

    2015-12-15

    Salmonella typhimurium and Staphylococcus aureus are most common causes of food-associated disease. A Raman based biosensor was developed for S. typhimurium and S. aureus detection simultaneously. The biosensor was based on nanoparticles enhanced Raman intensity and the specific recognition of aptamer. The Raman signal probe and the capture probe are built. Gold nanoparticles (GNPs) modified with Raman molecules (Mercaptobenzoic acid and 5,5'-Dithiobis(2-nitrobenzoic acid)) and aptamer are used as the signal probe for S. typhimurium and S. aureus, respectively. Fe3O4 magnetic gold nanoparticles (MGNPs) immobilized with both aptamer of S. typhimurium and S. aureus are used as the capture probe. When S. typhimurium and S. aureus are added in the reaction system, the capture probe will capture the target bacteria through the specific binding effect of aptamer. And then the signal probe will be connected to the bacteria also by the effect of aptamer to form the sandwich like detection structure. The Raman intensified spectrum was measured to quantify S. typhimurium and S. aureus. Under optimal conditions, the SERS intensity of MBA at 1582 cm(-1) are used to measure S. typhimurium (y=186.4762+704.8571x, R(2)=0.9921) and the SERS intensity of DNTB at 1333 cm(-1) are used to measure S. aureus (y=135.2381+211.4286x, R(2)=0.9946) in the range of 10(2)-10(7) cfu mL(-1). The LOD is 35 cfu mL(-1) for S. aureus and 15 cfu mL(-1) for S. typhimurium. This method is simple and rapid, results in high sensitivity and specificity, and can be used to detect actual samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Rapid and Reliable HPLC Method for the Simultaneous Determination of Dihydroxyacetone, Methylglyoxal and 5-Hydroxymethylfurfural in Leptospermum Honeys.

    Directory of Open Access Journals (Sweden)

    Matthew Pappalardo

    Full Text Available A reliable determination of dihydroxyacetone, methylglyoxal and 5-hydroxymethylfurfural is essential to establishing the commercial value and antimicrobial potential of honeys derived from the Leptospermum species endemic to Australia and New Zealand. We report a robust method for quantitation of all three compounds in a single HPLC run. Honey samples (n = 6 that are derivatized with o-(2,3,4,5,6-Pentafluorobenzyl hydroxylamine were quantitated against a stable anisole internal standard. Linear regression analysis was performed using calibration standards for each compound (n = 6 and results indicated a high degree of accuracy (R2 = 0.999 for this method. The reliability of some commercial methylglyoxal solutions were found to be questionable. Effective quantitation of methylglyoxal content in honey is critical for researchers and industry, and the use of some commercial standards may bias data. Two accurate methylglyoxal standards are proposed, including a commercial standard and a derivative that can be prepared within the laboratory.

  5. Reliability Study Regarding the Use of Histogram Similarity Methods for Damage Detection

    Directory of Open Access Journals (Sweden)

    Nicoleta Gillich

    2013-01-01

    Full Text Available The paper analyses the reliability of three dissimilarity estimators to compare histograms, as support for a frequency-based damage detection method, able to identify structural changes in beam-like structures. First a brief presentation of the own developed damage detection method is made, with focus on damage localization. It consists actually in comparing a histogram derived from measurement results, with a large series of histograms, namely the damage location indexes for all locations along the beam, obtained by calculus. We tested some dissimilarity estimators like the Minkowski-form Distances, the Kullback-Leibler Divergence and the Histogram Intersection and found the Minkowski Distance as the method providing best results. It was tested for numerous locations, using real measurement results and with results artificially debased by noise, proving its reliability.

  6. Indian program for development of technologies relevant to reliable, non-intrusive, concealed-contraband detection

    International Nuclear Information System (INIS)

    Auluck, S.K.H.

    2007-01-01

    Generating capability for reliable, non-intrusive detection of concealed-contraband, particularly, organic contraband like explosives and narcotics, has become a national priority. This capability spans a spectrum of technologies. If a technology mission addressing the needs of a highly sophisticated technology like PFNA is set up, the capabilities acquired would be adequate to meet the requirements of many other sets of technologies. This forms the background of the Indian program for development of technologies relevant to reliable, non-intrusive, concealed contraband detection. One of the central themes of the technology development programs would be modularization of the neutron source and detector technologies, so that common elements can be combined in different ways for meeting a variety of application requirements. (author)

  7. Scenario based approach to structural damage detection and its value in a risk and reliability perspective

    DEFF Research Database (Denmark)

    Hovgaard, Mads Knude; Hansen, Jannick Balleby; Brincker, Rune

    2013-01-01

    A scenario- and vibration based structural damage detection method is demonstrated though simulation. The method is Finite Element (FE) based. The value of the monitoring is calculated using structural reliability theory. A high cycle fatigue crack propagation model is assumed as the damage mecha......- and without monitoring. Monte Carlo Sampling (MCS) is used to estimate the probabilities and the tower of an onshore NREL 5MW wind turbine is given as a calculation case......A scenario- and vibration based structural damage detection method is demonstrated though simulation. The method is Finite Element (FE) based. The value of the monitoring is calculated using structural reliability theory. A high cycle fatigue crack propagation model is assumed as the damage...

  8. NDE reliability and probability of detection (POD) evolution and paradigm shift

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Surendra [NDE Engineering, Materials and Process Engineering, Honeywell Aerospace, Phoenix, AZ 85034 (United States)

    2014-02-18

    The subject of NDE Reliability and POD has gone through multiple phases since its humble beginning in the late 1960s. This was followed by several programs including the important one nicknamed “Have Cracks – Will Travel” or in short “Have Cracks” by Lockheed Georgia Company for US Air Force during 1974–1978. This and other studies ultimately led to a series of developments in the field of reliability and POD starting from the introduction of fracture mechanics and Damaged Tolerant Design (DTD) to statistical framework by Bernes and Hovey in 1981 for POD estimation to MIL-STD HDBK 1823 (1999) and 1823A (2009). During the last decade, various groups and researchers have further studied the reliability and POD using Model Assisted POD (MAPOD), Simulation Assisted POD (SAPOD), and applying Bayesian Statistics. All and each of these developments had one objective, i.e., improving accuracy of life prediction in components that to a large extent depends on the reliability and capability of NDE methods. Therefore, it is essential to have a reliable detection and sizing of large flaws in components. Currently, POD is used for studying reliability and capability of NDE methods, though POD data offers no absolute truth regarding NDE reliability, i.e., system capability, effects of flaw morphology, and quantifying the human factors. Furthermore, reliability and POD have been reported alike in meaning but POD is not NDE reliability. POD is a subset of the reliability that consists of six phases: 1) samples selection using DOE, 2) NDE equipment setup and calibration, 3) System Measurement Evaluation (SME) including Gage Repeatability and Reproducibility (Gage R and R) and Analysis Of Variance (ANOVA), 4) NDE system capability and electronic and physical saturation, 5) acquiring and fitting data to a model, and data analysis, and 6) POD estimation. This paper provides an overview of all major POD milestones for the last several decades and discuss rationale for using

  9. A novel approach for reliable detection of cathepsin S activities in mouse antigen presenting cells.

    Science.gov (United States)

    Steimle, Alex; Kalbacher, Hubert; Maurer, Andreas; Beifuss, Brigitte; Bender, Annika; Schäfer, Andrea; Müller, Ricarda; Autenrieth, Ingo B; Frick, Julia-Stefanie

    2016-05-01

    Cathepsin S (CTSS) is a eukaryotic protease mostly expressed in professional antigen presenting cells (APCs). Since CTSS activity regulation plays a role in the pathogenesis of various autoimmune diseases like multiple sclerosis, atherosclerosis, Sjögren's syndrome and psoriasis as well as in cancer progression, there is an ongoing interest in the reliable detection of cathepsin S activity. Various applications have been invented for specific detection of this enzyme. However, most of them have only been shown to be suitable for human samples, do not deliver quantitative results or the experimental procedure requires technical equipment that is not commonly available in a standard laboratory. We have tested a fluorogen substrate, Mca-GRWPPMGLPWE-Lys(Dnp)-DArg-NH2, that has been described to specifically detect CTSS activities in human APCs for its potential use for mouse samples. We have modified the protocol and thereby offer a cheap, easy, reproducible and quick activity assay to detect CTSS activities in mouse APCs. Since most of basic research on CTSS is performed in mice, this method closes a gap and offers a possibility for reliable and quantitative CTSS activity detection that can be performed in almost every laboratory. Copyright © 2016. Published by Elsevier B.V.

  10. Testing effort dependent software reliability model for imperfect debugging process considering both detection and correction

    International Nuclear Information System (INIS)

    Peng, R.; Li, Y.F.; Zhang, W.J.; Hu, Q.P.

    2014-01-01

    This paper studies the fault detection process (FDP) and fault correction process (FCP) with the incorporation of testing effort function and imperfect debugging. In order to ensure high reliability, it is essential for software to undergo a testing phase, during which faults can be detected and corrected by debuggers. The testing resource allocation during this phase, which is usually depicted by the testing effort function, considerably influences not only the fault detection rate but also the time to correct a detected fault. In addition, testing is usually far from perfect such that new faults may be introduced. In this paper, we first show how to incorporate testing effort function and fault introduction into FDP and then develop FCP as delayed FDP with a correction effort. Various specific paired FDP and FCP models are obtained based on different assumptions of fault introduction and correction effort. An illustrative example is presented. The optimal release policy under different criteria is also discussed

  11. Reliable Grid Condition Detection and Control of Single-Phase Distributed Power Generation Systems

    DEFF Research Database (Denmark)

    Ciobotaru, Mihai

    standards addressed to the grid-connected systems will harmonize the combination of the DPGS and the classical power plants. Consequently, the major tasks of this thesis were to develop new grid condition detection techniques and intelligent control in order to allow the DPGS not only to deliver power...... to the utility grid but also to sustain it. This thesis was divided into two main parts, namely "Grid Condition Detection" and "Control of Single-Phase DPGS". In the first part, the main focus was on reliable Phase Locked Loop (PLL) techniques for monitoring the grid voltage and on grid impedance estimation...... techniques. Additionally, a new technique for detecting the islanding mode has been developed and successfully tested. In the second part, the main reported research was concentrated around adaptive current controllers based on the information provided by the grid condition detection techniques. To guarantee...

  12. Simultaneous determination of psychotropic drugs in human urine by capillary electrophoresis with electrochemiluminescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Li Jianguo [Key Laboratory of Analytical Chemistry for Life Science (Education Ministry of China), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093 (China); School of Chemistry and Chemical Engineering, Suzhou University, Suzhou 215006 (China); Zhao Fengjuan [Key Laboratory of Analytical Chemistry for Life Science (Education Ministry of China), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093 (China); Ju Huangxian [Key Laboratory of Analytical Chemistry for Life Science (Education Ministry of China), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093 (China)]. E-mail: hxju@nju.edu.cn

    2006-08-04

    Amitriptyline, doxepin and chlorpromazine are often used as psychotropic drugs in treatment of the various mental diseases, and are also partly excreted by kidney. This work developed a simple, selective and sensitive method for their simultaneous monitoring in human urine using capillary electrophoresis coupled with electrochemiluminescence (ECL) detection based on end-column ECL reaction of tris-(2,2'-bipyridyl)ruthenium(II) with aliphatic tertiary amino moieties. Acetone was used as an additive to the running buffer to obtain their absolute separation. Under optimized conditions the proposed method displayed a linear range from 5.0 to 800 ng mL{sup -1} for the three drugs with the correlation coefficients more than 0.995 (n = 8). Their limits of detection were 0.8 ng mL{sup -1} (3.6 fg), 1.0 ng mL{sup -1} (4.5 fg) and 1.5 ng mL{sup -1} (6.8 fg) at a signal to noise ratio of 3, respectively. The relative standard deviations for five determinations of 20 ng mL{sup -1} amitriptyline, doxepin and chlorpromazine were 1.7%, 4.2% and 3.6%, respectively. For practical application an extract step with 90:10 heptane/ethyl acetate (v/v) was performed to eliminate the influence of ionic strength in sample. The recoveries of amitriptyline, doxepin and chlorpromazine at different levels in human urine were between 83% and 93%, which showed that the method was valuable in clinical and biochemical laboratories for monitoring amitriptyline, doxepin and chlorpromazine.

  13. Simultaneous determination of inorganic anions and cations by supercritical fluid chromatography using evaporative light scattering detection.

    Science.gov (United States)

    Foulon, Catherine; Di Giulio, Pauline; Lecoeur, Marie

    2018-01-26

    Supercritical fluid chromatography (SFC) is commonly used for the analysis of non-polar compounds, but remains poorly explored for the separation of polar and ionized molecules. In this paper, SFC has been investigated for the separation of 14 inorganic ions sampled in aqueous solutions. Four polar stationary phases were first screened using CO 2 -methanol-based mobile phases containing water or different acidic or basic additives, in order to select the most efficient conditions for the simultaneous retention of inorganic cations and anions and to favor their detection using evaporative light scattering detector (ELSD). Orthogonal selectivity was obtained depending on the stationary phase used: whereas anions are less retained on HILIC stationary phase, 2-ethylpyridine (2-EP) stationary phase exhibits strong interaction for anions. Best results were obtained under gradient elution mode using a 2-EP stationary phase and by adding 0.2% triethylamine in the CO 2 -methanol-based mobile phase. The composition of the injection solvent was also investigated. The results showed that a methanolic sample containing a percentage of water not exceeding 20% does not affect the analytical performances obtained on 2-EP. Moreover, the presence of triethylamine in the injection solvent contributes to eliminate peaks shoulders. Among the 14 inorganic ions tested, three cations (Li + , Ca 2+ and Mg 2+ ) and five anions (Cl - , Br - , NO 3 - , I - , SCN - ) were totally resolved in 15 min. NO 3 - and NO 2 - still coeluted in the final optimized conditions. The other investigated ions were either strongly retained on the stationary phase or not detected by the ELSD. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Is sequential cranial ultrasound reliable for detection of white matter injury in very preterm infants?

    International Nuclear Information System (INIS)

    Leijser, Lara M.; Steggerda, Sylke J.; Walther, Frans J.; Wezel-Meijler, Gerda van; Bruine, Francisca T. de; Grond, Jeroen van der

    2010-01-01

    Cranial ultrasound (cUS) may not be reliable for detection of diffuse white matter (WM) injury. Our aim was to assess in very preterm infants the reliability of a classification system for WM injury on sequential cUS throughout the neonatal period, using magnetic resonance imaging (MRI) as reference standard. In 110 very preterm infants (gestational age <32 weeks), serial cUS during admission (median 8, range 4-22) and again around term equivalent age (TEA) and a single MRI around TEA were performed. cUS during admission were assessed for presence of WM changes, and contemporaneous cUS and MRI around TEA additionally for abnormality of lateral ventricles. Sequential cUS (from birth up to TEA) and MRI were classified as normal/mildly abnormal, moderately abnormal, or severely abnormal, based on a combination of findings of the WM and lateral ventricles. Predictive values of the cUS classification were calculated. Sequential cUS were classified as normal/mildly abnormal, moderately abnormal, and severely abnormal in, respectively, 22%, 65%, and 13% of infants and MRI in, respectively, 30%, 52%, and 18%. The positive predictive value of the cUS classification for the MRI classification was high for severely abnormal WM (0.79) but lower for normal/mildly abnormal (0.67) and moderately abnormal (0.64) WM. Sequential cUS during the neonatal period detects severely abnormal WM in very preterm infants but is less reliable for mildly and moderately abnormal WM. MRI around TEA seems needed to reliably detect WM injury in very preterm infants. (orig.)

  15. A fast and reliable method for simultaneous waveform, amplitude and latency estimation of single-trial EEG/MEG data.

    Directory of Open Access Journals (Sweden)

    Wouter D Weeda

    Full Text Available The amplitude and latency of single-trial EEG/MEG signals may provide valuable information concerning human brain functioning. In this article we propose a new method to reliably estimate single-trial amplitude and latency of EEG/MEG signals. The advantages of the method are fourfold. First, no a-priori specified template function is required. Second, the method allows for multiple signals that may vary independently in amplitude and/or latency. Third, the method is less sensitive to noise as it models data with a parsimonious set of basis functions. Finally, the method is very fast since it is based on an iterative linear least squares algorithm. A simulation study shows that the method yields reliable estimates under different levels of latency variation and signal-to-noise ratioÕs. Furthermore, it shows that the existence of multiple signals can be correctly determined. An application to empirical data from a choice reaction time study indicates that the method describes these data accurately.

  16. Simultaneous determination of isoniazid and p-aminosalicylic acid by capillary electrophoresis using chemiluminescence detection.

    Science.gov (United States)

    Zhang, Xinfeng; Xuan, Yuelan; Sun, Aimin; Lv, Yi; Hou, Xiandeng

    2009-01-01

    It was found that isoniazid (ISO) or p-aminosalicylic acid (PAS) could enhance the chemiluminescence (CL) emission from Cu (II)-luminol-hydrogen peroxide system, and the increased chemiluminescence signals were proportional to their concentrations, respectively. Based on this phenomenon, a chemiluminescence method coupled to capillary electrophoresis (CE) was established for simultaneous determination of ISO and PAS. The CE conditions including running buffer and running voltage were investigated in detail. The effects of the pH of H(2)O(2) solution and the concentrations of luminol, H(2)O(2) and Cu (II) on the CL signal were also investigated carefully. Under the optimized conditions, the analysis could be accomplished within 10 min, with the limits of detection of 0.3 microg mL(-1) for ISO and 1.1 microg mL(-1) for PAS, corresponding to 7.2 and 26.4 pg per injection (24 nL), respectively. Finally, the method was validated by determining the two analytes in pharmaceutical preparation and spiked human serum samples. The results of pharmaceutical tablet analysis were in good agreement with the labeled amounts. The recoveries for ISO and PAS in human serum were in the range of 92-104% and 90-113%, respectively. Copyright 2008 John Wiley & Sons, Ltd.

  17. Simultaneous Observation Data of GB-SAR/PiSAR to Detect Flooding in an Urban Area

    Directory of Open Access Journals (Sweden)

    Manabu Watanabe

    2010-01-01

    Full Text Available We analyzed simultaneous observation data with ground-based synthetic aperture radar (GB-SAR and airborne SAR (PiSAR over a flood test site at which a simple house was constructed in a field. The PiSAR σ∘ under flood condition was 0.9 to 3.4 dB higher than that under nonflood condition. GB-SAR gives high spatial resolution as we could identify a single scattering component and a double bounce component from the house. GB-SAR showed that the σ∘ difference between the flooding and nonflooding conditions came from the double bounce scattering. We also confirm that the entropy is a sensitive parameter in the eigenvalue decomposition parameters, if the scattering process is dominated by the double bounce scattering. We conclude that σ∘ and entropy are a good parameter to be used to detect flooding, not only in agricultural and forest regions, but also in urban areas. We also conclude that GB-SAR is a powerful tool to supplement satellite and airborne observation, which has a relatively low spatial resolution.

  18. Simultaneous Observation Data of GB-SAR/PiSAR to Detect Flooding in an Urban Area

    Directory of Open Access Journals (Sweden)

    Shimada Masanobu

    2010-01-01

    Full Text Available Abstract We analyzed simultaneous observation data with ground-based synthetic aperture radar (GB-SAR and airborne SAR (PiSAR over a flood test site at which a simple house was constructed in a field. The PiSAR under flood condition was 0.9 to 3.4 dB higher than that under nonflood condition. GB-SAR gives high spatial resolution as we could identify a single scattering component and a double bounce component from the house. GB-SAR showed that the difference between the flooding and nonflooding conditions came from the double bounce scattering. We also confirm that the entropy is a sensitive parameter in the eigenvalue decomposition parameters, if the scattering process is dominated by the double bounce scattering. We conclude that and entropy are a good parameter to be used to detect flooding, not only in agricultural and forest regions, but also in urban areas. We also conclude that GB-SAR is a powerful tool to supplement satellite and airborne observation, which has a relatively low spatial resolution.

  19. FIRST SIMULTANEOUS DETECTION OF MOVING MAGNETIC FEATURES IN PHOTOSPHERIC INTENSITY AND MAGNETIC FIELD DATA

    International Nuclear Information System (INIS)

    Lim, Eun-Kyung; Yurchyshyn, Vasyl; Goode, Philip

    2012-01-01

    The formation and the temporal evolution of a bipolar moving magnetic feature (MMF) was studied with high-spatial and temporal resolution. The photometric properties were observed with the New Solar Telescope at Big Bear Solar Observatory using a broadband TiO filter (705.7 nm), while the magnetic field was analyzed using the spectropolarimetric data obtained by Hinode. For the first time, we observed a bipolar MMF simultaneously in intensity images and magnetic field data, and studied the details of its structure. The vector magnetic field and the Doppler velocity of the MMF were also studied. A bipolar MMF with its positive polarity closer to the negative penumbra formed, accompanied by a bright, filamentary structure in the TiO data connecting the MMF and a dark penumbral filament. A fast downflow (≤2 km s –1 ) was detected at the positive polarity. The vector magnetic field obtained from the full Stokes inversion revealed that a bipolar MMF has a U-shaped magnetic field configuration. Our observations provide a clear intensity counterpart of the observed MMF in the photosphere, and strong evidence of the connection between the MMF and the penumbral filament as a serpentine field.

  20. Simultaneous detection and analysis of optical and ultraviolet broad emission lines in quasars at z 2.2

    Science.gov (United States)

    Bisogni, S.; di Serego Alighieri, S.; Goldoni, P.; Ho, L. C.; Marconi, A.; Ponti, G.; Risaliti, G.

    2017-06-01

    We studied the spectra of six z 2.2 quasars obtained with the X-shooter spectrograph at the Very Large Telescope. The redshift of these sources and the X-shooter's spectral coverage allow us to cover the rest of the spectral range 1200-7000 Å for the simultaneous detection of optical and ultraviolet lines emitted by the broad-line region. Simultaneous measurements, avoiding issues related to quasars variability, help us understand the connection between the different broad-line region line profiles generally used as virial estimators of black hole masses in quasars. The goal of this work is to compare the different emission lines for each object to check on the reliability of Hα, Mg II and C iv with respect to Hβ. Hα and Mg II linewidths correlate well with Hβ, while C iv shows a poorer correlation, due to the presence of strong blueshifts and asymmetries in the profile. We compared our sample with the only other two whose spectra were taken with the same instrument and for all examined lines our results are in agreement with the ones obtained with X-shooter at z 1.5-1.7. We finally evaluate C III] as a possible substitute of C iv in the same spectral range and find that its behaviour is more coherent with those of the other lines: we believe that, when a high quality spectrum such as the ones we present is available and a proper modelization with the Fe II and Fe III emissions is performed, it is more appropriate to use this line than that of C iv if not corrected for the contamination by non-virialized components. Based on observations collected at the European Organisation for Astronomical Research in the Southern Hemisphere, Chile, under programme 086.B-0320(A).The reduced spectra (FITS files) are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (http://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/603/A1

  1. One-step multiplex quantitative RT-PCR for the simultaneous detection of viroids and phytoplasmas of pome fruit trees.

    Science.gov (United States)

    Malandraki, Ioanna; Varveri, Christina; Olmos, Antonio; Vassilakos, Nikon

    2015-03-01

    A one-step multiplex real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) based on TaqMan chemistry was developed for the simultaneous detection of Pear blister canker viroid and Apple scar skin viroid along with universal detection of phytoplasmas, in pome trees. Total nucleic acids (TNAs) extraction was performed according to a modified CTAB protocol. Primers and TaqMan MGB probes for specific detection of the two viroids were designed in this study, whereas for phytoplasma detection published universal primers and probe were used, with the difference that the later was modified to carry a MGB quencher. The pathogens were detected simultaneously in 10-fold serial dilutions of TNAs from infected plant material into TNAs of healthy plant up to dilutions 10(-5) for viroids and 10(-4) for phytoplasmas. The multiplex real-time assay was at least 10 times more sensitive than conventional protocols for viroid and phytoplasma detection. Simultaneous detection of the three targets was achieved in composite samples at least up to a ratio of 1:100 triple-infected to healthy tissue, demonstrating that the developed assay has the potential to be used for rapid and massive screening of viroids and phytoplasmas of pome fruit trees in the frame of certification schemes and surveys. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. reliability reliability

    African Journals Online (AJOL)

    eobe

    Corresponding author, Tel: +234-703. RELIABILITY .... V , , given by the code of practice. However, checks must .... an optimization procedure over the failure domain F corresponding .... of Concrete Members based on Utility Theory,. Technical ...

  3. Simultaneous detection and identification of four cherry viruses by two step multiplex RT-PCR with an internal control of plant nad5 mRNA.

    Science.gov (United States)

    Noorani, Md Salik; Awasthi, Prachi; Sharma, Maheshwar Prasad; Ram, Raja; Zaidi, Aijaz Asgar; Hallan, Vipin

    2013-10-01

    A multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed and standardized for the simultaneous detection of four cherry viruses: Cherry virus A (CVA, Genus; Capillovirus), Cherry necrotic rusty mottle virus (CNRMV, unassigned species of the Betaflexiviridae), Little cherry virus 1 (LChV-1, Genus; Closterovirus) and Prunus necrotic ringspot virus (PNRSV, Genus; Ilarvirus) with nad5 as plant internal control. A reliable and quick method for total plant RNA extraction from pome and stone fruit trees was also developed. To minimize primer dimer formation, a single antisense primer for CVA and CNRMV was used. A mixture of random hexamer and oligo (dT) primer was used for cDNA synthesis, which was highly suited and economic for multiplexing. All four viruses were detected successfully by mRT-PCR in artificially created viral RNA mixture and field samples of sweet cherry. The identity of the viruses was confirmed by sequencing. The assay could detect above viruses in diluted cDNA (10(-4)) and RNA (10(-3), except PNRSV which was detected only till ten times lesser dilution). The developed mRT-PCR will not only be useful for the detection of viruses from single or multiple infections of sweet cherry plants but also for other stone and pome fruits. The developed method will be therefore quite helpful for virus indexing, plant quarantine and certification programs. This is the first report for the simultaneous detection of four cherry viruses by mRT-PCR. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Quantitative Analysis of Humectants in Tobacco Products Using Gas Chromatography (GC with Simultaneous Mass Spectrometry (MSD and Flame Ionization Detection (FID

    Directory of Open Access Journals (Sweden)

    Rainey CL

    2014-12-01

    Full Text Available This paper describes the modification of an existing gas chromatographic (GC method to incorporate simultaneous mass spectrometric (MSD and flame ionization detection (FID into the analysis of tobacco humectants. Glycerol, propylene glycol, and triethylene glycol were analyzed in tobacco labeled as roll-your-own (RYO, cigar, cigarette, moist snuff, and hookah tobacco. Tobacco was extracted in methanol containing 1,3-butanediol (internal standard, filtered, and separated on a 15 m megabore DB-Wax column. Post-column flow was distributed using a microfluidic splitter between the MSD and FID for simultaneous detection. The limits of detection for the FID detector were 0.5 μg/mL (propylene glycol and triethylene glycol and 0.25 μg/mL (glycerol with a linear range of 2-2000 μg/mL (propylene glycol and triethylene glycol and 1-4000 μg/mL (glycerol. The limits of detection for the MSD detector were 2 μg/mL (propylene glycol and triethylene glycol and 4 μg/mL (glycerol with a linear range of 20-2000 μg/mL (propylene glycol and triethylene glycol and 40-4000 μg/mL (glycerol. Significant improvement in the sensitivity of the MSD can be achieved by employing selective ion monitoring (SIM detection mode. Although a high degree of correlation was observed between the results from FID and MSD analyses, marginal chromatographic resolution between glycerol and triethylene glycol limits the applicability of FID to samples containing low levels of both of these humectants. Utilizing MSD greatly improves the reliability of quantitative results because compensation for inadequate chromatographic resolution can be accomplished with mass selectivity in detection.

  5. A sensitive whole-cell biosensor for the simultaneous detection of a broad-spectrum of toxic heavy metal ions.

    Science.gov (United States)

    Cerminati, S; Soncini, F C; Checa, S K

    2015-04-07

    Bacterial biosensors are simple, cost-effective and efficient analytical tools for detecting bioavailable heavy metals in the environment. This work presents the design, construction and calibration of a novel whole-cell fluorescent biosensory device that, simultaneously and with high sensitivity, reports the presence of toxic mercury, lead, cadmium and/or gold ions in aqueous samples. This bio-reporter can be easily applied as an immediate alerting tool for detecting the presence of harmful pollutants in drinking water.

  6. Multiplex PCR Assay for Identification of Six Different Staphylococcus spp. and Simultaneous Detection of Methicillin and Mupirocin Resistance

    OpenAIRE

    Campos-Peña, E.; Martín-Nuñez, E.; Pulido-Reyes, G.; Martín-Padrón, J.; Caro-Carrillo, E.; Donate-Correa, J.; Lorenzo-Castrillejo, I.; Alcoba-Flórez, J.; Machín, F.; Méndez-Alvarez, S.

    2014-01-01

    We describe a new, efficient, sensitive, and fast single-tube multiple-PCR protocol for the identification of the most clinically significant Staphylococcus spp. and the simultaneous detection of the methicillin and mupirocin resistance loci. The protocol identifies at the species level isolates belonging to S. aureus, S. epidermidis, S. haemolyticus, S. hominis, S. lugdunensis, and S. saprophyticus.

  7. Simultaneous detection of multiple DNA targets by integrating dual-color graphene quantum dot nanoprobes and carbon nanotubes.

    Science.gov (United States)

    Qian, Zhaosheng; Shan, Xiaoyue; Chai, Lujing; Chen, Jianrong; Feng, Hui

    2014-12-01

    Simultaneous detection of multiple DNA targets was achieved based on a biocompatible graphene quantum dots (GQDs) and carbon nanotubes (CNTs) platform through spontaneous assembly between dual-color GQD-based probes and CNTs and subsequently self-recognition between DNA probes and targets. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Reliability and Minimum Detectable Change of Temporal-Spatial, Kinematic, and Dynamic Stability Measures during Perturbed Gait.

    Directory of Open Access Journals (Sweden)

    Christopher A Rábago

    Full Text Available Temporal-spatial, kinematic variability, and dynamic stability measures collected during perturbation-based assessment paradigms are often used to identify dysfunction associated with gait instability. However, it remains unclear which measures are most reliable for detecting and tracking responses to perturbations. This study systematically determined the between-session reliability and minimum detectable change values of temporal-spatial, kinematic variability, and dynamic stability measures during three types of perturbed gait. Twenty young healthy adults completed two identical testing sessions two weeks apart, comprised of an unperturbed and three perturbed (cognitive, physical, and visual walking conditions in a virtual reality environment. Within each session, perturbation responses were compared to unperturbed walking using paired t-tests. Between-session reliability and minimum detectable change values were also calculated for each measure and condition. All temporal-spatial, kinematic variability and dynamic stability measures demonstrated fair to excellent between-session reliability. Minimal detectable change values, normalized to mean values ranged from 1-50%. Step width mean and variability measures demonstrated the greatest response to perturbations with excellent between-session reliability and low minimum detectable change values. Orbital stability measures demonstrated specificity to perturbation direction and sensitivity with excellent between-session reliability and low minimum detectable change values. We observed substantially greater between-session reliability and lower minimum detectable change values for local stability measures than previously described which may be the result of averaging across trials within a session and using velocity versus acceleration data for reconstruction of state spaces. Across all perturbation types, temporal-spatial, orbital and local measures were the most reliable measures with the

  9. Mixed-substrate (glycerol tributyrate and fibrin) zymography for simultaneous detection of lipolytic and proteolytic enzymes on a single gel.

    Science.gov (United States)

    Choi, Nack-Shick; Choi, Jong Hyun; Kim, Bo-Hye; Han, Yun-Jon; Kim, Joong Su; Lee, Seung-Goo; Song, Jae Jun

    2009-06-01

    A new zymography method for simultaneous detection of two different enzymatic activities (lipolytic and proteolytic) using a single SDS-containing or native-conformation gel and a mixed-substrate (glycerol tributyrate and fibrin) (MS)(1) gel was developed. After routine electrophoresis, SDS in the gel was removed by treatment with Triton X-100. Gel proteins were electrotransferred to the MS gel. To visualize lipolytic activity, the MS gel was incubated at 37 degrees C (for 6 or 24 h) until clear bands against an opaque background were observed. To detect proteolytic activity, the same MS gel was stained with Coomassie brilliant blue. Using this method, we show that six lipolytic enzymes from Staphylococcus pasteuri NJ-1 and four proteolytic enzymes from two Bacillus strains, B. licheniformis DJ-2 and B. licheniformis NJ-5, isolated from soil, can be simultaneously detected.

  10. Simultaneous quantitative determination of six active components in traditional Chinese medicinal preparation Cerebralcare Granule® by RP-HPLC coupled with diode array detection for quality control.

    Science.gov (United States)

    Wang, Xiang-yang; Ma, Xiao-hui; Li, Wei; Chu, Yang; Guo, Jia-hua; Zhou, Shui-ping; Zhu, Yong-hong

    2014-09-01

    A simple, accurate and reliable method for the simultaneous separation and determination of six active components (protocatechuic acid, chlorogenic acid, caffeic acid, paeoniflorin, ferulic acid and rosmarinic acid) in traditional Chinese medicinal preparation Cerebralcare Granule(®) (CG) was developed using reverse-phase high-performance liquid chromatography coupled with diode array detector detection. The chromatographic separation was performed on a Hypersil GOLD C18 column with aqueous formic acid (0.1%, v/v) and acetonitrile as mobile phase at a flow rate of 0.2 ml/min at 30 °C. Because of the different UV characteristics of these components, change detection wavelength method was used for quantitative analysis. All of the analytes showed good linearity (r > 0.9992). The established method showed good precision and relative standard deviations (%) for intra-day and inter-day variations of 0.15-1.81 and 0.11-1.98%, respectively. The validated method was successfully applied to the simultaneously determination of six active components in CG from different batches. © The Author [2013]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  11. Pharyngeal pH alone is not reliable for the detection of pharyngeal reflux events: A study with oesophageal and pharyngeal pH-impedance monitoring

    Science.gov (United States)

    Desjardin, Marie; Roman, Sabine; des Varannes, Stanislas Bruley; Gourcerol, Guillaume; Coffin, Benoit; Ropert, Alain; Mion, François

    2013-01-01

    Background Pharyngeal pH probes and pH-impedance catheters have been developed for the diagnosis of laryngo-pharyngeal reflux. Objective To determine the reliability of pharyngeal pH alone for the detection of pharyngeal reflux events. Methods 24-h pH-impedance recordings performed in 45 healthy subjects with a bifurcated probe for detection of pharyngeal and oesophageal reflux events were reviewed. Pharyngeal pH drops to below 4 and 5 were analysed for the simultaneous occurrence of pharyngeal reflux, gastro-oesophageal reflux, and swallows, according to impedance patterns. Results Only 7.0% of pharyngeal pH drops to below 5 identified with impedance corresponded to pharyngeal reflux, while 92.6% were related to swallows and 10.2 and 13.3% were associated with proximal and distal gastro-oesophageal reflux events, respectively. Of pharyngeal pH drops to below 4, 13.2% were related to pharyngeal reflux, 87.5% were related to swallows, and 18.1 and 21.5% were associated with proximal and distal gastro-oesophageal reflux events, respectively. Conclusions This study demonstrates that pharyngeal pH alone is not reliable for the detection of pharyngeal reflux and that adding distal oesophageal pH analysis is not helpful. The only reliable analysis should take into account impedance patterns demonstrating the presence of pharyngeal reflux event preceded by a distal and proximal reflux event within the oesophagus. PMID:24917995

  12. Simultaneous quantitative detection of multiple tumor markers with a rapid and sensitive multicolor quantum dots based immunochromatographic test strip.

    Science.gov (United States)

    Wang, Chunying; Hou, Fei; Ma, Yicai

    2015-06-15

    A novel multicolor quantum dots (QDs) based immunochromatographic test strip (ICTS) was developed for simultaneous quantitative detection of multiple tumor markers, by utilizing alpha fetoprotein (AFP) and carcinoembryonic antigen (CEA) as models. The immunosensor could realize simultaneous quantitative detection of tumor markers with only one test line and one control line on the nitrocellulose membrane (NC membrane) due to the introduction of multicolor QDs. In this method, a mixture of mouse anti-AFP McAb and mouse anti-CEA McAb was coated on NC membrane as test line and goat anti-mouse IgG antibody was coated as control line. Anti-AFP McAb-QDs546 conjugates and anti-CEA McAb-QDs620 conjugates were mixed and applied to the conjugate pad. Simultaneous quantitative detection of multiple tumor markers was achieved by detecting the fluorescence intensity of captured QDs labels on test line and control line using a test strip reader. Under the optimum conditions, AFP and CEA could be detected as low as 3 ng/mL and 2 ng/mL in 15 min with a sample volume of 80 μL, and no obvious cross-reactivity was observed. The immunosensor was validated with 130 clinical samples and in which it exhibited high sensitivity (93% for AFP and 87% for CEA) and specificity (94% for AFP and 97% for CEA). The immunosensor also demonstrated high recoveries (87.5-113% for AFP and 90-97.3% for CEA) and low relative standard deviations (RSDs) (2.8-6.2% for AFP and 4.9-9.6% for CEA) when testing spiked human serum. This novel multicolor QDs based ICTS provides an easy and rapid, simultaneous quantitative detecting strategy for point-of-care testing of tumor markers. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. MOA-2010-BLG-311: A PLANETARY CANDIDATE BELOW THE THRESHOLD OF RELIABLE DETECTION

    International Nuclear Information System (INIS)

    Yee, J. C.; Hung, L.-W.; Gould, A.; Gaudi, B. S.; Bond, I. A.; Allen, W.; Monard, L. A. G.; Albrow, M. D.; Fouqué, P.; Dominik, M.; Tsapras, Y.; Udalski, A.; Zellem, R.; Bos, M.; Christie, G. W.; DePoy, D. L.; Dong, Subo; Drummond, J.; Gorbikov, E.; Han, C.

    2013-01-01

    We analyze MOA-2010-BLG-311, a high magnification (A max > 600) microlensing event with complete data coverage over the peak, making it very sensitive to planetary signals. We fit this event with both a point lens and a two-body lens model and find that the two-body lens model is a better fit but with only Δχ 2 ∼ 80. The preferred mass ratio between the lens star and its companion is q = 10 –3.7±0.1 , placing the candidate companion in the planetary regime. Despite the formal significance of the planet, we show that because of systematics in the data the evidence for a planetary companion to the lens is too tenuous to claim a secure detection. When combined with analyses of other high-magnification events, this event helps empirically define the threshold for reliable planet detection in high-magnification events, which remains an open question.

  14. MOA-2010-BLG-311: A PLANETARY CANDIDATE BELOW THE THRESHOLD OF RELIABLE DETECTION

    Energy Technology Data Exchange (ETDEWEB)

    Yee, J. C.; Hung, L.-W.; Gould, A.; Gaudi, B. S. [Department of Astronomy, Ohio State University, 140 West 18th Avenue, Columbus, OH 43210 (United States); Bond, I. A. [Institute for Information and Mathematical Sciences, Massey University, Private Bag 102-904, Auckland 1330 (New Zealand); Allen, W. [Vintage Lane Observatory, Blenheim (New Zealand); Monard, L. A. G. [Bronberg Observatory, Centre for Backyard Astrophysics, Pretoria (South Africa); Albrow, M. D. [Department of Physics and Astronomy, University of Canterbury, Private Bag 4800, Christchurch 8020 (New Zealand); Fouque, P. [IRAP, CNRS, Universite de Toulouse, 14 avenue Edouard Belin, F-31400 Toulouse (France); Dominik, M. [SUPA, University of St. Andrews, School of Physics and Astronomy, North Haugh, St. Andrews, KY16 9SS (United Kingdom); Tsapras, Y. [Las Cumbres Observatory Global Telescope Network, 6740B Cortona Drive, Goleta, CA 93117 (United States); Udalski, A. [Warsaw University Observatory, Al. Ujazdowskie 4, 00-478 Warszawa (Poland); Zellem, R. [Department of Planetary Sciences/LPL, University of Arizona, 1629 East University Boulevard, Tucson, AZ 85721 (United States); Bos, M. [Molehill Astronomical Observatory, North Shore City, Auckland (New Zealand); Christie, G. W. [Auckland Observatory, P.O. Box 24-180, Auckland (New Zealand); DePoy, D. L. [Department of Physics, Texas A and M University, 4242 TAMU, College Station, TX 77843-4242 (United States); Dong, Subo [Institute for Advanced Study, Einstein Drive, Princeton, NJ 08540 (United States); Drummond, J. [Possum Observatory, Patutahi (New Zealand); Gorbikov, E. [School of Physics and Astronomy, Raymond and Beverley Sackler Faculty of Exact Sciences, Tel-Aviv University, Tel Aviv 69978 (Israel); Han, C., E-mail: liweih@astro.ucla.edu, E-mail: rzellem@lpl.arizona.edu, E-mail: tim.natusch@aut.ac.nz [Department of Physics, Chungbuk National University, 410 Seongbong-Rho, Hungduk-Gu, Chongju 371-763 (Korea, Republic of); Collaboration: muFUN Collaboration; MOA Collaboration; OGLE Collaboration; PLANET Collaboration; RoboNet Collaboration; MiNDSTEp Consortium; and others

    2013-05-20

    We analyze MOA-2010-BLG-311, a high magnification (A{sub max} > 600) microlensing event with complete data coverage over the peak, making it very sensitive to planetary signals. We fit this event with both a point lens and a two-body lens model and find that the two-body lens model is a better fit but with only {Delta}{chi}{sup 2} {approx} 80. The preferred mass ratio between the lens star and its companion is q = 10{sup -3.7{+-}0.1}, placing the candidate companion in the planetary regime. Despite the formal significance of the planet, we show that because of systematics in the data the evidence for a planetary companion to the lens is too tenuous to claim a secure detection. When combined with analyses of other high-magnification events, this event helps empirically define the threshold for reliable planet detection in high-magnification events, which remains an open question.

  15. Analysis of non simultaneous common mode failures. Application to the reliability assessment of the decay heat removal of the RNR 1500 project

    International Nuclear Information System (INIS)

    Natta, M.; Bloch, M.

    1991-01-01

    The experience with the LMFBR PHENIX has shown many cases of failures on identical and redundant components, which were close in time but not simultaneous and due to the same causes such as a design error, an unappropriate material, corrosion, ... Since the decay heat removal (DHR) must be assured for a long period after shutdown of the reactor, the overall reliability of the DHR system depends much on this type of successive failures by common mode causes, for which the usual β factor methods are not appropriate since they imply that the several failures are simultaneous. In this communication, two methods will be presented. The first one was used to assess the reliability of the DHR system of the RNR 1500 project. In this method, one modelize the occurrence of successive failures on n identical files by a sudden jump of the failure rate from the value λ attributed to the first failure to the value λ' attributed to the (n-1) still available files. This method leads to a quite natural quantification of the interest of diversity for highly redundant systems. For the RNR 1500 project where, in case of the loss of normal DHR path through the steam generators, the decay heat is removed by four separated sodium loops of 26 MW unit capacity in forced convection, the probabilistic assessment shows that it is necessary to diversify the sodium-sodium heat exchanger in order to fullfil the upper limit of 10 -7 /year for the probability of failure of DHR. A separate assessment for the main sequence leading to DHR loss was performed using a different method in which the successive failures are interpreted as a premature end of life, the lifetimes being directly used as random variables. This Monte-Carlo type method, which can be applied to any type of lifetime distribution, leads to results consistent to those obtained with the first one

  16. HPLC Method for Simultaneous Quantitative Detection of Quercetin and Curcuminoids in Traditional Chinese Medicines

    Directory of Open Access Journals (Sweden)

    Lee Fung Ang

    2014-12-01

    Full Text Available Objectives: Quercetin and curcuminoids are important bioactive compounds found in many herbs. Previously reported high performance liquid chromatography ultraviolet (HPLC-UV methods for the detection of quercetin and curcuminoids have several disadvantages, including unsatisfactory separation times and lack of validation according the standard guidelines of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. Methods: A rapid, specific, reversed phase, HPLC-UV method with an isocratic elution of acetonitrile and 2% v/v acetic acid (40% : 60% v/v (pH 2.6 at a flow rate of 1.3 mL/minutes, a column temperature of 35°C, and ultraviolet (UV detection at 370 nm was developed. The method was validated and applied to the quantification of different types of market available Chinese medicine extracts, pills and tablets. Results: The method allowed simultaneous determination of quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin in the concentration ranges of 0.00488 ─ 200 μg/mL, 0.625 ─ 320 μg/mL, 0.07813 ─ 320 μg/mL and 0.03906 ─ 320 μg/mL, respectively. The limits of detection and quantification, respectively, were 0.00488 and 0.03906 μg/mL for quercetin, 0.62500 and 2.50000 μg/mL for bisdemethoxycurcumin, 0.07813 and 0.31250 μg/mL for demethoxycurcumin, and 0.03906 and 0.07813 μg/mL for curcumin. The percent relative intra day standard deviation (% RSD values were 0.432 ─ 0.806 μg/mL, 0.576 ─ 0.723 μg/ mL, 0.635 ─ 0.752 μg/mL and 0.655 ─ 0.732 μg/mL for quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin, respectively, and those for intra day precision were 0.323 ─ 0.968 μg/mL, 0.805 ─ 0.854 μg/mL, 0.078 ─ 0.844 μg/mL and 0.275 ─ 0.829 μg/mL, respectively. The intra day accuracies were 99.589% ─ 100.821%, 98.588% ─ 101.084%, 9.289% ─ 100.88%, and 98.292% ─ 101.022% for quercetin, bisdemethoxycurcumin

  17. HPLC method for simultaneous quantitative detection of quercetin and curcuminoids in traditional chinese medicines.

    Science.gov (United States)

    Ang, Lee Fung; Yam, Mun Fei; Fung, Yvonne Tan Tze; Kiang, Peh Kok; Darwin, Yusrida

    2014-12-01

    Quercetin and curcuminoids are important bioactive compounds found in many herbs. Previously reported high performance liquid chromatography ultraviolet (HPLC-UV) methods for the detection of quercetin and curcuminoids have several disadvantages, including unsatisfactory separation times and lack of validation according the standard guidelines of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. A rapid, specific, reversed phase, HPLC-UV method with an isocratic elution of acetonitrile and 2% v/v acetic acid (40% : 60% v/v) (pH 2.6) at a flow rate of 1.3 mL/minutes, a column temperature of 35°C, and ultraviolet (UV) detection at 370 nm was developed. The method was validated and applied to the quantification of different types of market available Chinese medicine extracts, pills and tablets. The method allowed simultaneous determination of quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin in the concentration ranges of 0.00488 ─ 200 μg/mL, 0.625 ─ 320 μg/mL, 0.07813 ─ 320 μg/mL and 0.03906 ─ 320 μg/mL, respectively. The limits of detection and quantification, respectively, were 0.00488 and 0.03906 μg/mL for quercetin, 0.62500 and 2.50000 μg/mL for bisdemethoxycurcumin, 0.07813 and 0.31250 μg/mL for demethoxycurcumin, and 0.03906 and 0.07813 μg/mL for curcumin. The percent relative intra day standard deviation (% RSD) values were 0.432 ─ 0.806 μg/mL, 0.576 ─ 0.723 μg/mL, 0.635 ─ 0.752 μg/mL and 0.655 ─ 0.732 μg/mL for quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin, respectively, and those for intra day precision were 0.323 ─ 0.968 μg/mL, 0.805 ─ 0.854 μg/mL, 0.078 ─ 0.844 μg/mL and 0.275 ─ 0.829 μg/mL, respectively. The intra day accuracies were 99.589% ─ 100.821%, 98.588% ─ 101.084%, 9.289% ─ 100.88%, and 98.292% ─ 101.022% for quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin, respectively, and the

  18. Simultaneous detection of three pome fruit tree viruses by one-step multiplex quantitative RT-PCR.

    Science.gov (United States)

    Malandraki, Ioanna; Beris, Despoina; Isaioglou, Ioannis; Olmos, Antonio; Varveri, Christina; Vassilakos, Nikon

    2017-01-01

    A one-step multiplex real-time reverse transcription polymerase chain reaction (RT-qPCR) based on TaqMan probes was developed for the simultaneous detection of Apple mosaic virus (ApMV), Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) in total RNA of pome trees extracted with a CTAB method. The sensitivity of the method was established using in vitro synthesized viral transcripts serially diluted in RNA from healthy, virus-tested (negative) pome trees. The three viruses were simultaneously detected up to a 10-4 dilution of total RNA from a naturally triple-infected apple tree prepared in total RNA of healthy apple tissue. The newly developed RT-qPCR assay was at least one hundred times more sensitive than conventional single RT-PCRs. The assay was validated with 36 field samples for which nine triple and 11 double infections were detected. All viruses were detected simultaneously in composite samples at least up to the ratio of 1:150 triple-infected to healthy pear tissue, suggesting the assay has the capacity to examine rapidly a large number of samples in pome tree certification programs and surveys for virus presence.

  19. Simultaneous detection of six stone fruit viruses by non-isotopic molecular hybridization using a unique riboprobe or 'polyprobe'.

    Science.gov (United States)

    Herranz, M Carmen; Sanchez-Navarro, Jesus A; Aparicio, Frederic; Pallás, Vicente

    2005-03-01

    A new strategy for the simultaneous detection of plant viruses by molecular hybridization has been developed. Two, four or six viral sequences were fused in tandem and transcribed to render unique riboprobes and designated as 'polyprobes'. The 'polyprobe four' (poly 4) covered the four ilarviruses affecting stone fruit trees including apple mosaic virus (ApMV), prunus necrotic ringspot virus (PNRSV), prune dwarf virus (PDV), and American plum line pattern virus (APLPV) whereas the 'polyprobe two' (poly 2) was designed to detect simultaneously, plum pox virus (PPV) and apple chlorotic leaf spot virus (ACLSV), the two more important viruses affecting these trees. Finally, a 'polyprobe six' (poly 6) was generated to detect any of the six viruses. The three polyprobes were comparable to the individual riboprobes in terms of end-point dilution limit and specificity. The validation of the new simultaneous detection strategy was confirmed by the analysis of 46 field samples from up to seven different hosts collected from 10 different geographical areas.

  20. Simultaneous detection of ultraviolet B-induced DNA damage using capillary electrophoresis with laser-induced fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Guthrie, Jeffrey W., E-mail: jeff.guthrie@emich.edu; Limmer, Robert T.; Brooks, Eric A.; Wisnewski, Chelsea C.; Loggins-Davis, Nnekia D.; Bouzid, Abderraouf

    2015-01-01

    Highlights: • CE–LIF was developed for simultaneous detection of UV-induced DNA photoproducts. • Fluorescent quantum dot reporters enabled detection of small amounts of photoproducts. • Photoproducts were detected after 65 J m{sup −2} of fluence from a UVB lamp in ∼6 ng of DNA. • Natural sunlight induced cyclobutane pyrimidine dimers after only 15 min of exposure. - Abstract: An immunoassay based on CE–LIF was developed for the simultaneous detection of cyclobutane pyrimidine dimers (CPDs) and pyrimidine 6-4 pyrimidone photoproducts (6-4PPs) in genomic DNA irradiated with UVB or natural sunlight. Human cells were first exposed to varying amounts of UVB or natural sunlight to induce DNA damage. Genomic DNA was extracted and incubated with anti-CPD and anti-6-4PP primary antibodies attached to secondary antibodies with a fluorescent quantum dot (QD) reporter that emitted either red or yellow fluorescence. CE was used to separate the unbound antibodies from those bound to the photoproducts, and LIF with appropriate optical filters was used to separate the fluorescence signals from each QD to individual photomultiplier tubes for simultaneous photoproduct detection. Using this strategy, photoproducts were detected from ∼6 ng (200 ng μL{sup −1}) of DNA under a low UVB fluence of 65 J m{sup −2} for CPDs or 195 J m{sup −2} for 6-4PPs. This assay was also the first to demonstrate the detection of CPDs in human cells after only 15 min of irradiation under natural sunlight.

  1. Simultaneous detection of ultraviolet B-induced DNA damage using capillary electrophoresis with laser-induced fluorescence

    International Nuclear Information System (INIS)

    Guthrie, Jeffrey W.; Limmer, Robert T.; Brooks, Eric A.; Wisnewski, Chelsea C.; Loggins-Davis, Nnekia D.; Bouzid, Abderraouf

    2015-01-01

    Highlights: • CE–LIF was developed for simultaneous detection of UV-induced DNA photoproducts. • Fluorescent quantum dot reporters enabled detection of small amounts of photoproducts. • Photoproducts were detected after 65 J m −2 of fluence from a UVB lamp in ∼6 ng of DNA. • Natural sunlight induced cyclobutane pyrimidine dimers after only 15 min of exposure. - Abstract: An immunoassay based on CE–LIF was developed for the simultaneous detection of cyclobutane pyrimidine dimers (CPDs) and pyrimidine 6-4 pyrimidone photoproducts (6-4PPs) in genomic DNA irradiated with UVB or natural sunlight. Human cells were first exposed to varying amounts of UVB or natural sunlight to induce DNA damage. Genomic DNA was extracted and incubated with anti-CPD and anti-6-4PP primary antibodies attached to secondary antibodies with a fluorescent quantum dot (QD) reporter that emitted either red or yellow fluorescence. CE was used to separate the unbound antibodies from those bound to the photoproducts, and LIF with appropriate optical filters was used to separate the fluorescence signals from each QD to individual photomultiplier tubes for simultaneous photoproduct detection. Using this strategy, photoproducts were detected from ∼6 ng (200 ng μL −1 ) of DNA under a low UVB fluence of 65 J m −2 for CPDs or 195 J m −2 for 6-4PPs. This assay was also the first to demonstrate the detection of CPDs in human cells after only 15 min of irradiation under natural sunlight

  2. Multiplex PCR assay for the simultaneous detection of C. perfringens, P. aeruginosa and K. pneumoniae

    Directory of Open Access Journals (Sweden)

    Pradeepkiran Jangampalli Adi

    2015-11-01

    Results and conclusions: Through this approach, the above pathogens were detected simultaneously with high specificity in pure cultures and from the blood and urine samples. The results were correlated with normal diagnostic process, and proved to be more sensitive and specific diagnostic technique in the simultaneous detection of C. perfringens, P. aeruginosa and K. pneumoniae.

  3. Visual acuity measures do not reliably detect childhood refractive error--an epidemiological study.

    Directory of Open Access Journals (Sweden)

    Lisa O'Donoghue

    Full Text Available PURPOSE: To investigate the utility of uncorrected visual acuity measures in screening for refractive error in white school children aged 6-7-years and 12-13-years. METHODS: The Northern Ireland Childhood Errors of Refraction (NICER study used a stratified random cluster design to recruit children from schools in Northern Ireland. Detailed eye examinations included assessment of logMAR visual acuity and cycloplegic autorefraction. Spherical equivalent refractive data from the right eye were used to classify significant refractive error as myopia of at least 1DS, hyperopia as greater than +3.50DS and astigmatism as greater than 1.50DC, whether it occurred in isolation or in association with myopia or hyperopia. RESULTS: Results are presented from 661 white 12-13-year-old and 392 white 6-7-year-old school-children. Using a cut-off of uncorrected visual acuity poorer than 0.20 logMAR to detect significant refractive error gave a sensitivity of 50% and specificity of 92% in 6-7-year-olds and 73% and 93% respectively in 12-13-year-olds. In 12-13-year-old children a cut-off of poorer than 0.20 logMAR had a sensitivity of 92% and a specificity of 91% in detecting myopia and a sensitivity of 41% and a specificity of 84% in detecting hyperopia. CONCLUSIONS: Vision screening using logMAR acuity can reliably detect myopia, but not hyperopia or astigmatism in school-age children. Providers of vision screening programs should be cognisant that where detection of uncorrected hyperopic and/or astigmatic refractive error is an aspiration, current UK protocols will not effectively deliver.

  4. A Method for Improving Reliability of Radiation Detection using Deep Learning Framework

    International Nuclear Information System (INIS)

    Chang, Hojong; Kim, Tae-Ho; Han, Byunghun; Kim, Hyunduk; Kim, Ki-duk

    2017-01-01

    Radiation detection is essential technology for overall field of radiation and nuclear engineering. Previously, technology for radiation detection composes of preparation of the table of the input spectrum to output spectrum in advance, which requires simulation of numerous predicted output spectrum with simulation using parameters modeling the spectrum. In this paper, we propose new technique to improve the performance of radiation detector. The software in the radiation detector has been stagnant for a while with possible intrinsic error of simulation. In the proposed method, to predict the input source using output spectrum measured by radiation detector is performed using deep neural network. With highly complex model, we expect that the complex pattern between data and the label can be captured well. Furthermore, the radiation detector should be calibrated regularly and beforehand. We propose a method to calibrate radiation detector using GAN. We hope that the power of deep learning may also reach to radiation detectors and make huge improvement on the field. Using improved radiation detector, the reliability of detection would be confident, and there are many tasks remaining to solve using deep learning in nuclear engineering society.

  5. Simultaneous detection of the 160-min pulsations of the Sun with two radiotelescopes

    International Nuclear Information System (INIS)

    Nesterov, N.S.; Urno, S.; Kotov, V.A.

    1983-01-01

    The differential (center-to-limb) radio briqhtness of the quet Sun was measured on June 22, 1981 simultaneously in Crimea at 13.5-mm wavelength and in Finland at 8-mm wavelength. Both independent sets of observations have shown the presence of synchronous variation of the solar radioemission with the 160-min period

  6. Simultaneous detection of water-soluble vitamins using the High Performance Liquid Chromatography (HPLC - a review

    Directory of Open Access Journals (Sweden)

    Rosemond Godbless Dadzie

    2014-01-01

    Full Text Available The water-soluble vitamins (WSV: ascorbic acid (vitamin C, thiamine (B1, riboflavin (B2, niacin (B3, panthothenic acid (B5, pyridoxine, and pyridoxal (B6, folic acid (B9, biotin(B8 , and B12 are very essential in the diet of humankind. As a result of ever increasing pressures from both consumers and legal enforcers, to specify accurately nutritive compositions of WSV that are present in food materials, many researchers have attempted to fill this niche through the provision of highly sensitive and rapid high performance liquid chromatography (HPLC procedures. In view of the health benefits of WSV, a replete of HPLC methods have been developed for simultaneous determination of their contents in nature and fortified food samples, nutritional supplements, as well as blood plasmas. The rate of losses of these vitamins during food processing and analysis, in addition to their transient dynamics, presents complexities in developing a highly sensitive HPLC procedure for their simultaneous separations and assays. This review critically assesses the different HPLC procedures developed by researchers and available in the open literature for simultaneous determination of water-soluble vitamins (WSV in dried tropical fruits materials. The study revealed that not a single chromatographic run developed by researchers can simultaneously elute all the WSV at a time. However, the HPLC procedures that are capable of determining all the WSV were coupled with electrospray ionization mass spectroscopy (ESI-MS, thus making the set-up expensive.

  7. Designing a reliable leak bio-detection system for natural gas pipelines

    International Nuclear Information System (INIS)

    Batzias, F.A.; Siontorou, C.G.; Spanidis, P.-M.P.

    2011-01-01

    Monitoring of natural gas (NG) pipelines is an important task for economical/safety operation, loss prevention and environmental protection. Timely and reliable leak detection of gas pipeline, therefore, plays a key role in the overall integrity management for the pipeline system. Owing to the various limitations of the currently available techniques and the surveillance area that needs to be covered, the research on new detector systems is still thriving. Biosensors are worldwide considered as a niche technology in the environmental market, since they afford the desired detector capabilities at low cost, provided they have been properly designed/developed and rationally placed/networked/maintained by the aid of operational research techniques. This paper addresses NG leakage surveillance through a robust cooperative/synergistic scheme between biosensors and conventional detector systems; the network is validated in situ and optimized in order to provide reliable information at the required granularity level. The proposed scheme is substantiated through a knowledge based approach and relies on Fuzzy Multicriteria Analysis (FMCA), for selecting the best biosensor design that suits both, the target analyte and the operational micro-environment. This approach is illustrated in the design of leak surveying over a pipeline network in Greece.

  8. Designing a reliable leak bio-detection system for natural gas pipelines

    Energy Technology Data Exchange (ETDEWEB)

    Batzias, F.A., E-mail: fbatzi@unipi.gr [Univ. Piraeus, Dept. Industrial Management and Technology, Karaoli and Dimitriou 80, 18534 Piraeus (Greece); Siontorou, C.G., E-mail: csiontor@unipi.gr [Univ. Piraeus, Dept. Industrial Management and Technology, Karaoli and Dimitriou 80, 18534 Piraeus (Greece); Spanidis, P.-M.P., E-mail: pspani@asprofos.gr [Asprofos Engineering S.A, El. Venizelos 284, 17675 Kallithea (Greece)

    2011-02-15

    Monitoring of natural gas (NG) pipelines is an important task for economical/safety operation, loss prevention and environmental protection. Timely and reliable leak detection of gas pipeline, therefore, plays a key role in the overall integrity management for the pipeline system. Owing to the various limitations of the currently available techniques and the surveillance area that needs to be covered, the research on new detector systems is still thriving. Biosensors are worldwide considered as a niche technology in the environmental market, since they afford the desired detector capabilities at low cost, provided they have been properly designed/developed and rationally placed/networked/maintained by the aid of operational research techniques. This paper addresses NG leakage surveillance through a robust cooperative/synergistic scheme between biosensors and conventional detector systems; the network is validated in situ and optimized in order to provide reliable information at the required granularity level. The proposed scheme is substantiated through a knowledge based approach and relies on Fuzzy Multicriteria Analysis (FMCA), for selecting the best biosensor design that suits both, the target analyte and the operational micro-environment. This approach is illustrated in the design of leak surveying over a pipeline network in Greece.

  9. Designing a reliable leak bio-detection system for natural gas pipelines.

    Science.gov (United States)

    Batzias, F A; Siontorou, C G; Spanidis, P-M P

    2011-02-15

    Monitoring of natural gas (NG) pipelines is an important task for economical/safety operation, loss prevention and environmental protection. Timely and reliable leak detection of gas pipeline, therefore, plays a key role in the overall integrity management for the pipeline system. Owing to the various limitations of the currently available techniques and the surveillance area that needs to be covered, the research on new detector systems is still thriving. Biosensors are worldwide considered as a niche technology in the environmental market, since they afford the desired detector capabilities at low cost, provided they have been properly designed/developed and rationally placed/networked/maintained by the aid of operational research techniques. This paper addresses NG leakage surveillance through a robust cooperative/synergistic scheme between biosensors and conventional detector systems; the network is validated in situ and optimized in order to provide reliable information at the required granularity level. The proposed scheme is substantiated through a knowledge based approach and relies on Fuzzy Multicriteria Analysis (FMCA), for selecting the best biosensor design that suits both, the target analyte and the operational micro-environment. This approach is illustrated in the design of leak surveying over a pipeline network in Greece. Copyright © 2010 Elsevier B.V. All rights reserved.

  10. Diagnostic reliability of 3.0-T MRI for detecting osseous abnormalities of the temporomandibular joint.

    Science.gov (United States)

    Sawada, Kunihiko; Amemiya, Toshihiko; Hirai, Shigenori; Hayashi, Yusuke; Suzuki, Toshihiro; Honda, Masahiko; Sisounthone, Johnny; Matsumoto, Kunihito; Honda, Kazuya

    2018-01-01

    We compared the diagnostic reliability of 3.0-T magnetic resonance imaging (MRI) for detection of osseous abnormalities of the temporomandibular joint (TMJ) with that of the gold standard, cone-beam computed tomography (CBCT). Fifty-six TMJs were imaged with CBCT and MRI, and images of condyles and fossae were independently assessed for the presence of osseous abnormalities. The accuracy, sensitivity, and specificity of 3.0-T MRI were 0.88, 1.0, and 0.73, respectively, in condyle evaluation and 0.91, 0.75, and 0.95 in fossa evaluation. The McNemar test showed no significant difference (P > 0.05) between MRI and CBCT in the evaluation of osseous abnormalities in condyles and fossae. The present results indicate that 3.0-T MRI is equal to CBCT in the diagnostic evaluation of osseous abnormalities of the mandibular condyle.

  11. Development of duplex RT-PCR-ELISA for the simultaneous detection of hepatitis A virus and hepatitis E virus.

    Science.gov (United States)

    Tahk, Hongmin; Lee, Min Hwa; Lee, Kang Bum; Cheon, Doo-Sung; Choi, Changsun

    2011-07-01

    This study aimed to develop a specific and sensitive duplex reverse transcription polymerase chain reaction enzyme-linked immunosorbent assay (duplex RT-PCR-ELISA) for hepatitis A virus (HAV) and hepatitis E virus (HEV). Duplex RT-PCR-ELISA could detect and differentiate HAV and HEV with specific probes. When ELISA technique was used to detect probe-bound RT-PCR products, duplex RT-PCR-ELISA could detect as little as 0.1 ng/μL HAV and HEV from clinical samples. Human norovirus, enterovirus, poliovirus, murine norovirus and feline calicivirus were used for the specificity test; all were negative. Therefore duplex RT-PCR-ELISA can be used for the simultaneous detection of HAV and HEV in contaminated fecal samples. Copyright © 2011 Elsevier B.V. All rights reserved.

  12. Multiplex hydrolysis probe real-time PCR for simultaneous detection of hepatitis A virus and hepatitis E virus.

    Science.gov (United States)

    Qiu, Feng; Cao, Jingyuan; Su, Qiudong; Yi, Yao; Bi, Shengli

    2014-05-30

    Detection of hepatitis viral infections has traditionally relied on the circulating antibody test using the enzyme-linked immunosorbent assay. However, multiplex real-time PCR has been increasingly used for a variety of viral nucleic acid detections and has proven to be superior to traditional methods. Hepatitis A virus (HAV) and hepatitis E virus (HEV) are the major causes of acute hepatitis worldwide; both HAV and HEV infection are a main public health problem. In the present study, a one-step multiplex reverse transcriptase quantitative polymerase chain reaction assay using hydrolysis probes was developed for simultaneously detecting HAV and HEV. This novel detection system proved specific to the target viruses, to be highly sensitive and to be applicable to clinical sera samples, making it useful for rapid, accurate and feasible identification of HAV and HEV.

  13. Modifying a standard method allows simultaneous extraction of RNA and protein, enabling detection of enzymes in the rat retina with low expressions and protein levels.

    Science.gov (United States)

    Agardh, Elisabet; Gustavsson, Carin; Hagert, Per; Nilsson, Marie; Agardh, Carl-David

    2006-02-01

    The aim of the study was to evaluate messenger RNA and protein expression in limited amounts of tissue with low protein content. The Chomczynski method was used for simultaneous extraction of RNA, and protein was modified in the protein isolation step. Template mass and cycling time for the complementary DNA synthesis step of real-time reverse transcription-polymerase chain reaction (RT-PCR) for analysis of catalase, copper/zinc superoxide dismutase, manganese superoxide dismutase, the catalytic subunit of glutamylcysteine ligase, glutathione peroxidase 1, and the endogenous control cyclophilin B (CypB) were optimized before PCR. Polymerase chain reaction accuracy and efficacy were demonstrated by calculating the regression (R2) values of the separate amplification curves. Appropriate antibodies, blocking buffers, and running conditions were established for Western blot, and protein detection and multiplex assays with CypB were performed for each target. During the extraction procedure, the protein phase was dissolved in a modified washing buffer containing 0.1% sodium dodecyl sulfate, followed by ultrafiltration. Enzyme expression on real-time RT-PCR was accomplished with high reliability and reproducibility (R2, 0.990-0.999), and all enzymes except for glutathione peroxidase 1 were detectable in individual retinas on Western blot. Western blot multiplexing with CypB was possible for all targets. In conclusion, connecting gene expression directly to protein levels in the individual rat retina was possible by simultaneous extraction of RNA and protein. Real-time RT-PCR and Western blot allowed accurate detection of retinal protein expressions and levels.

  14. Rapid and sensitive approach to simultaneous detection of genomes of hepatitis A, B, C, D and E viruses.

    Science.gov (United States)

    Kodani, Maja; Mixson-Hayden, Tonya; Drobeniuc, Jan; Kamili, Saleem

    2014-10-01

    Five viruses have been etiologically associated with viral hepatitis. Nucleic acid testing (NAT) remains the gold standard for diagnosis of viremic stages of infection. NAT methodologies have been developed for all hepatitis viruses; however, a NAT-based assay that can simultaneously detect all five viruses is not available. We designed TaqMan card-based assays for detection of HAV RNA, HBV DNA, HCV RNA, HDV RNA and HEV RNA. The performances of individual assays were evaluated on TaqMan Array Cards (TAC) for detecting five viral genomes simultaneously. Sensitivity and specificity were determined by testing 329 NAT-tested clinical specimens. All NAT-positive samples for HCV (n = 32), HDV (n = 28) and HEV (n = 14) were also found positive in TAC (sensitivity, 100%). Forty-three of 46 HAV-NAT positive samples were also positive in TAC (sensitivity, 94%), while 36 of 39 HBV-NAT positive samples were positive (sensitivity, 92%). No false-positives were detected for HBV (n = 32), HCV (n = 36), HDV (n = 30), and HEV (n = 31) NAT-negative samples (specificity 100%), while 38 of 41 HAV-NAT negative samples were negative by TAC (specificity 93%). TAC assay was concordant with corresponding individual NATs for hepatitis A-E viral genomes and can be used for their detection simultaneously. The TAC assay has potential for use in hepatitis surveillance, for screening of donor specimens and in outbreak situations. Wider availability of TAC-ready assays may allow for customized assays, for improving acute jaundice surveillance and for other purposes for which there is need to identify multiple pathogens rapidly. Published by Elsevier B.V.

  15. Simultaneous detection and differentiation of three genotypes of Brassica yellows virus by multiplex reverse transcription-polymerase chain reaction.

    Science.gov (United States)

    Zhang, Xiaoyan; Peng, Yanmei; Wang, Ying; Zhang, Zongying; Li, Dawei; Yu, Jialin; Han, Chenggui

    2016-11-22

    Brassica yellows virus (BrYV), proposed to be a new polerovirus species, three distinct genotypes (BrYV-A, BrYV-B and BrYV-C) have been described. This study was to develop a simple, rapid, sensitive, cost-effective method for simultaneous detection and differentiation of three genotypes of BrYV. In this study, a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed for simultaneous detection and differentiation of the three genotypes of BrYV. The three genotypes of BrYV and Tunip yellows virus (TuYV) could be differentiated simultaneously using six optimized specific oligonucleotide primers, including one universal primer for detecting BrYV, three BrYV genotype-specific primers, and a pair of primers for specific detection of TuYV. Primers were designed from conserved regions of each virus and their specificity was confirmed by sequencing PCR products. The mRT-PCR products were 278 bp for BrYV-A, 674 bp for BrYV-B, 505 bp for BrYV-C, and 205 bp for TuYV. Amplification of three target genotypes was optimized by increasing the PCR annealing temperatures to 62 °C. One to three fragments specific for the virus genotypes were simultaneously amplified from infected samples and identified by their specific molecular sizes in agarose gel electrophoresis. No specific products could be amplified from cDNAs of other viruses which could infect crucifer crops. Detection limits of the plasmids for multiplex PCR were 100 fg for BrYV-A and BrYV-B, 10 pg for BrYV-C, and 1 pg for TuYV, respectively. The mRT-PCR was applied successfully for detection of three BrYV genotypes from field samples collected in China. The simple, rapid, sensitive, and cost-effective mRT-PCR was developed successfully for detection and differentiation of the three genotypes of BrYV.

  16. Technical note: Simultaneous carotenoid and vitamin analysis of milk from total mixed ration-fed cows optimized for xanthophyll detection.

    Science.gov (United States)

    Stout, M A; Benoist, D M; Drake, M A

    2018-06-01

    Concentrations of retinol, α-tocopherol, and major carotenoids in dairy products are often determined simultaneously by liquid chromatography. These compounds have different polarity and solubility; thus, extracting them simultaneously can be difficult and inefficient. In milks with low carotenoid concentrations, the xanthophylls lutein and zeaxanthin may not be completely resolved using common extraction techniques. A simplified method was developed to optimize extraction efficiency and the limit of detection and limit of quantification (LoQ) of lutein and zeaxanthin in bovine milk without decreasing sensitivity to other vitamins or carotenoids. The developed method evaluates lutein, zeaxanthin, β-carotene, retinol, and α-tocopherol simultaneously by ultra-high performance liquid chromatography-photodiode array detection. Common saponification temperatures (40-60°C) and concentrations of KOH in water (10-50% KOH wt/vol) were evaluated. Multiple solvents were evaluated for optimal xanthophyll extraction (diethyl ether, dichloromethane, hexane, and tetrahydrofuran) following saponification. The limit of detection and LoQ were defined as 3:1 and 10:1 signal-to-noise ratio, respectively. All experiments were performed in triplicate. The optimal saponification procedure was a concentration of 25% KOH at either 40 or 50°C. Saponified extracts solubilized in solutions containing diethyl ether had greater concentrations of lutein- than hexane- or tetrahydrofuran-based solutions, with peak areas above LoQ values. The solution containing diethyl ether solubilized similar concentrations of retinol, α-tocopherol, and β-carotene when compared with other solutions. The proposed optimized method allows for the simultaneous determination of carotenoids from milk with increased lutein and zeaxanthin sensitivity without sacrificing recovery of retinol, α-tocopherol, and β-carotene. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights

  17. Photogrammetry: an accurate and reliable tool to detect thoracic musculoskeletal abnormalities in preterm infants.

    Science.gov (United States)

    Davidson, Josy; dos Santos, Amelia Miyashiro N; Garcia, Kessey Maria B; Yi, Liu C; João, Priscila C; Miyoshi, Milton H; Goulart, Ana Lucia

    2012-09-01

    To analyse the accuracy and reproducibility of photogrammetry in detecting thoracic abnormalities in infants born prematurely. Cross-sectional study. The Premature Clinic at the Federal University of São Paolo. Fifty-eight infants born prematurely in their first year of life. Measurement of the manubrium/acromion/trapezius angle (degrees) and the deepest thoracic retraction (cm). Digitised photographs were analysed by two blinded physiotherapists using a computer program (SAPO; http://SAPO.incubadora.fapesp.br) to detect shoulder elevation and thoracic retraction. Physical examinations performed independently by two physiotherapists were used to assess the accuracy of the new tool. Thoracic alterations were detected in 39 (67%) and in 40 (69%) infants by Physiotherapists 1 and 2, respectively (kappa coefficient=0.80). Using a receiver operating characteristic curve, measurement of the manubrium/acromion/trapezius angle and the deepest thoracic retraction indicated accuracy of 0.79 and 0.91, respectively. For measurement of the manubrium/acromion/trapezius angle, the Bland and Altman limits of agreement were -6.22 to 7.22° [mean difference (d)=0.5] for repeated measures by one physiotherapist, and -5.29 to 5.79° (d=0.75) between two physiotherapists. For thoracic retraction, the intra-rater limits of agreement were -0.14 to 0.18cm (d=0.02) and the inter-rater limits of agreement were -0.20 to -0.17cm (d=0.02). SAPO provided an accurate and reliable tool for the detection of thoracic abnormalities in preterm infants. Copyright © 2011 Chartered Society of Physiotherapy. Published by Elsevier Ltd. All rights reserved.

  18. Reliability of the exercise ECG in detecting silent ischemia in patients with prior myocardial infarction

    International Nuclear Information System (INIS)

    Yamagishi, Takashi; Matsuda, Yasuo; Satoh, Akira

    1991-01-01

    To assess the reliability of the exercise ECG in detecting silent ischemia, ECG results were compared with those of stress-redistribution thallium-201 single-photon emission computed tomography (SPECT) in 116 patients with prior myocardial infarction and in 20 normal subjects used as a control. The left ventricle (LV) was divided into 20 segmental images, which were scored blindly on a 5-point scale. The redistribution score was defined as thallium defect score of exercise subtracted by that of redistribution image and was used as a measure of amount of ischemic but viable myocardium. The upper limit of normal redistribution score (=4.32) was defined as mean+2 standard deviations derived from 20 normal subjects. Of 116 patients, 47 had the redistribution score above the normal range. Twenty-five (53%) of the 47 patients showed positive ECG response. Fourteen (20%) of the 69 patients, who had the normal redistribution score, showed positive ECG response. Thus, the ECG response had a sensitivity of 53% and a specificity of 80% in detecting transient ischemia. Furthermore, the 116 patients were subdivided into 4 groups according to the presence or absence of chest pain and ECG change during exercise. Fourteen patients showed both chest pain and ECG change and all these patients had the redistribution score above the normal range. Twenty-five patients showed ECG change without chest pain and 11 (44%) of the 25 patients had the abnormal redistribution. Three (43%) of 7 patients who showed chest pain without ECG change had the abnormal redistribution score. Of 70 patients who had neither chest pain nor ECG change, 19 (27%) had the redistribution score above the normal range. Thus, limitations exist in detecting silent ischemia by ECG in patients with a prior myocardial infarction, because the ECG response to the exercise test may have a low degree of sensitivity and a high degree of false positive and false negative results in detecting silent ischemia. (author)

  19. Prevent cervical cancer by screening with reliable human papillomavirus detection and genotyping

    International Nuclear Information System (INIS)

    Ge, Shichao; Gong, Bo; Cai, Xushan; Yang, Xiaoer; Gan, Xiaowei; Tong, Xinghai; Li, Haichuan; Zhu, Meijuan; Yang, Fengyun; Zhou, Hongrong; Hong, Guofan

    2012-01-01

    The incidence of cervical cancer is expected to rise sharply in China. A reliable routine human papillomavirus (HPV) detection and genotyping test to be supplemented by the limited Papanicolaou cytology facilities is urgently needed to help identify the patients with cervical precancer for preventive interventions. To this end, we evaluated a nested polymerase chain reaction (PCR) protocol for detection of HPV L1 gene DNA in cervicovaginal cells. The PCR amplicons were genotyped by direct DNA sequencing. In parallel, split samples were subjected to a Digene HC2 HPV test which has been widely used for “cervical cancer risk” screen. Of the 1826 specimens, 1655 contained sufficient materials for analysis and 657 were truly negative. PCR/DNA sequencing showed 674 infected by a single high-risk HPV, 188 by a single low-risk HPV, and 136 by multiple HPV genotypes with up to five HPV genotypes in one specimen. In comparison, the HC2 test classified 713 specimens as infected by high-risk HPV, and 942 as negative for HPV infections. The high-risk HC2 test correctly detected 388 (57.6%) of the 674 high-risk HPV isolates in clinical specimens, mislabeled 88 (46.8%) of the 188 low-risk HPV isolates as high-risk genotypes, and classified 180 (27.4%) of the 657 “true-negative” samples as being infected by high-risk HPV. It was found to cross-react with 20 low-risk HPV genotypes. We conclude that nested PCR detection of HPV followed by short target DNA sequencing can be used for screening and genotyping to formulate a paradigm in clinical management of HPV-related disorders in a rapidly developing economy

  20. A SIMULTANEOUS MULTI-PROBE DETECTION LABEL-FREE OPTICAL-RESOLUTION PHOTOACOUSTIC MICROSCOPY TECHNIQUE BASED ON MICROCAVITY TRANSDUCER

    Directory of Open Access Journals (Sweden)

    YONGBO WU

    2013-07-01

    Full Text Available We demonstrate the feasibility of simultaneous multi-probe detection for an optical-resolution photoacoustic microscopy (OR-PAM system. OR-PAM has elicited the attention of biomedical imaging researchers because of its optical absorption contrast and high spatial resolution with great imaging depth. OR-PAM allows label-free and noninvasive imaging by maximizing the optical absorption of endogenous biomolecules. However, given the inadequate absorption of some biomolecules, detection sensitivity at the same incident intensity requires improvement. In this study, a modulated continuous wave with power density less than 3 mW/cm2 (1/4 of the ANSI safety limit excited the weak photoacoustic (PA signals of biological cells. A microcavity transducer is developed based on the bulk modulus of gas five orders of magnitude lower than that of solid; air pressure variation is inversely proportional to cavity volume at the same temperature increase. Considering that a PA wave expands in various directions, detecting PA signals from different positions and adding them together can increase detection sensitivity and signal-to-noise ratio. Therefore, we employ four detectors to acquire tiny PA signals simultaneously. Experimental results show that the developed OR-PAM system allows the label-free imaging of cells with weak optical absorption.

  1. Detection of Trypanosoma cruzi in untreated chronic chagasic patients is improved by using three parasitological methods simultaneously.

    Science.gov (United States)

    Zulantay, Inés; Apt, Werner; Valencia, Claudio; Torres, Alberto; Saavedra, Miguel; Rodríguez, Jorge; Sandoval, Lea; Martínez, Gabriela; Thieme, Patricio; Sepúlveda, Eduardo

    2011-10-01

    This study compared three parasitological methods applied simultaneously in individuals with untreated chronic Chagas' disease in order to determine their individual and combined performances. From a total of 100 chronic chagasic patients from endemic areas of Chile, with informed consent, we extracted 2 mL of peripheral venous blood for PCR (PCR-B) and applied two xenodiagnosis (XD) boxes with seven uninfected Triatoma infestans nymphs each for microscopic examination and PCR of faecal samples of the triatomines fed on each patient (PCR-XD). The PCR-B and PCR-XD reactions were performed with oligonucleotides 121 and 122, which anneal to the four constant regions of the minicircles of Trypanosoma cruzi kinetoplasts. The 330 bp PCR product was analysed by electrophoresis in a 2% agarose gel and visualized by staining with ethidium bromide. PCR-B detected T. cruzi in 58% of the cases, while PCR-XD proved to be more sensitive than XD (67% versus 14%, respectively) (P = 0.0001). There was no difference between the detection power of PCR-B and PCR-XD (P = 0.222). The percentage detected as positive was much greater when the three tests were considered (84%) (P = 0.00001). The simultaneous application of more than one technique for the parasitological diagnosis of Chagas' disease in untreated individuals increases the possibility of detection of T. cruzi.

  2. Simultaneous detection of ultraviolet B-induced DNA damage using capillary electrophoresis with laser-induced fluorescence.

    Science.gov (United States)

    Guthrie, Jeffrey W; Limmer, Robert T; Brooks, Eric A; Wisnewski, Chelsea C; Loggins-Davis, Nnekia D; Bouzid, Abderraouf

    2015-01-01

    An immunoassay based on CE-LIF was developed for the simultaneous detection of cyclobutane pyrimidine dimers (CPDs) and pyrimidine 6-4 pyrimidone photoproducts (6-4PPs) in genomic DNA irradiated with UVB or natural sunlight. Human cells were first exposed to varying amounts of UVB or natural sunlight to induce DNA damage. Genomic DNA was extracted and incubated with anti-CPD and anti-6-4PP primary antibodies attached to secondary antibodies with a fluorescent quantum dot (QD) reporter that emitted either red or yellow fluorescence. CE was used to separate the unbound antibodies from those bound to the photoproducts, and LIF with appropriate optical filters was used to separate the fluorescence signals from each QD to individual photomultiplier tubes for simultaneous photoproduct detection. Using this strategy, photoproducts were detected from ∼6 ng (200 ng μL(-1)) of DNA under a low UVB fluence of 65 J m(-2) for CPDs or 195 J m(-2) for 6-4PPs. This assay was also the first to demonstrate the detection of CPDs in human cells after only 15 min of irradiation under natural sunlight. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Simultaneous Detection and Removal of Formaldehyde at Room Temperature: Janus Au@ZnO@ZIF-8 Nanoparticles

    Science.gov (United States)

    Wang, Dawei; Li, Zhiwei; Zhou, Jian; Fang, Hong; He, Xiang; Jena, Puru; Zeng, Jing-Bin; Wang, Wei-Ning

    2018-03-01

    The detection and removal of volatile organic compounds (VOCs) are of great importance to reduce the risk of indoor air quality concerns. This study reports the rational synthesis of a dual-functional Janus nanostructure and its feasibility for simultaneous detection and removal of VOCs. The Janus nanostructure was synthesized via an anisotropic growth method, composed of plasmonic nanoparticles, semiconductors, and metal organic frameworks (e.g., Au@ZnO@ZIF-8). It exhibits excellent selective detection to formaldehyde (HCHO, as a representative VOC) at room temperature over a wide range of concentrations (from 0.25 to 100 ppm), even in the presence of water and toluene molecules as interferences. In addition, HCHO was also found to be partially oxidized into non-toxic formic acid simultaneously with detection. The mechanism underlying this technology was unraveled by both experimental measurements and theoretical calculations: ZnO maintains the conductivity, while ZIF-8 improves the selective gas adsorption; the plasmonic effect of Au nanorods enhances the visible-light-driven photocatalysis of ZnO at room temperature. [Figure not available: see fulltext.

  4. Simultaneous detection of P300 and steady-state visually evoked potentials for hybrid brain-computer interface.

    Science.gov (United States)

    Combaz, Adrien; Van Hulle, Marc M

    2015-01-01

    We study the feasibility of a hybrid Brain-Computer Interface (BCI) combining simultaneous visual oddball and Steady-State Visually Evoked Potential (SSVEP) paradigms, where both types of stimuli are superimposed on a computer screen. Potentially, such a combination could result in a system being able to operate faster than a purely P300-based BCI and encode more targets than a purely SSVEP-based BCI. We analyse the interactions between the brain responses of the two paradigms, and assess the possibility to detect simultaneously the brain activity evoked by both paradigms, in a series of 3 experiments where EEG data are analysed offline. Despite differences in the shape of the P300 response between pure oddball and hybrid condition, we observe that the classification accuracy of this P300 response is not affected by the SSVEP stimulation. We do not observe either any effect of the oddball stimulation on the power of the SSVEP response in the frequency of stimulation. Finally results from the last experiment show the possibility of detecting both types of brain responses simultaneously and suggest not only the feasibility of such hybrid BCI but also a gain over pure oddball- and pure SSVEP-based BCIs in terms of communication rate.

  5. A novel GMO biosensor for rapid ultrasensitive and simultaneous detection of multiple DNA components in GMO products.

    Science.gov (United States)

    Huang, Lin; Zheng, Lei; Chen, Yinji; Xue, Feng; Cheng, Lin; Adeloju, Samuel B; Chen, Wei

    2015-04-15

    Since the introduction of genetically modified organisms (GMOs), there has been on-going and continuous concern and debates on the commercialization of products derived from GMOs. There is an urgent need for development of highly efficient analytical methods for rapid and high throughput screening of GMOs components, as required for appropriate labeling of GMO-derived foods, as well as for on-site inspection and import/export quarantine. In this study, we describe, for the first time, a multi-labeling based electrochemical biosensor for simultaneous detection of multiple DNA components of GMO products on the same sensing interface. Two-round signal amplification was applied by using both an exonuclease enzyme catalytic reaction and gold nanoparticle-based bio-barcode related strategies, respectively. Simultaneous multiple detections of different DNA components of GMOs were successfully achieved with satisfied sensitivity using this electrochemical biosensor. Furthermore, the robustness and effectiveness of the proposed approach was successfully demonstrated by application to various GMO products, including locally obtained and confirmed commercial GMO seeds and transgenetic plants. The proposed electrochemical biosensor demonstrated unique merits that promise to gain more interest in its use for rapid and on-site simultaneous multiple screening of different components of GMO products. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Simultaneous detection of metronidazole and chloramphenicol by differential pulse stripping voltammetry using a silver nanoparticles/sulfonate functionalized graphene modified glassy carbon electrode

    International Nuclear Information System (INIS)

    Zhai, Haiyun; Liang, Zhixian; Chen, Zuanguang; Wang, Haihang; Liu, Zhenping; Su, Zihao; Zhou, Qing

    2015-01-01

    Graphical abstract: Display Omitted -- Highlights: • A novel and reliable AgNPs/SF-GR modified glassy carbon electrode was constructed and characterized. • The AgNPs/SF-GR/GCE was successfully applied in the shrimp for simultaneous determination of MTZ and CAP. • Under optimized conditions, common substances such as UA, AA, DA and ion did not interfered in the electrode performance. • The modified electrode exhibited considerable sensitivity, stability and reproducibility. • This fabricated electrode achieved a satisfactory level compared with other electrodes toward MTZ and CAP. -- Abstract: A novel silver nanoparticles/sulfonated functionalized graphene modified glassy carbon electrode (AgNPs/SF-GR/GCE) was fabricated to determine chloramphenicol and metronidazole simultaneously. Taking advantage of sulfonic group, AgNPs were successfully electrodeposited on functionalized GR immobilized on the surface of a GCE. Scanning electron microscopy and energy spectrum analysis results confirmed that AgNPs were deposited on the functionalized GR film. Compared to the bare GCE or the pristine SF-GR modified electrode, AgNPs/SF-GR/GCE exhibited excellent electroreduction towards chloramphenicol and metronidazole. In addition, the two antibacterial drugs were separated completely in 0.10 M citric acid-sodium citrate buffer (pH 4.0) by differential pulse stripping voltammetry under optimum conditions. The cathodic current was linearly related with 0.02∼20.0 μM chloramphenicol and 0.10∼20.0 μM metronidazole, with the detection limits of 0.01 μM and 0.05 μM respectively. Furthermore, AgNPs/SF-GR/GCE was applied to the simultaneous determination of chloramphenicol and metronidazole in an aquatic product

  7. Quantitative multiplex assay for simultaneous detection and identification of Indiana and New Jersey serotypes of vesicular stomatitis virus

    DEFF Research Database (Denmark)

    Rasmussen, Thomas Bruun; Uttenthal, Åse; Fernandez, Jovita

    2005-01-01

    In order to establish a rapid and reliable system for the detection of vesicular stomatitis virus (VSV), we developed a quantitative reverse transcription-PCR assay for the detection, quantification, and differentiation of the major serotypes, VSV Indiana and VSV New Jersey, using a closed......-tube multiplex format. The detection system is based on the recently invented primer-probe energy transfer (PriProET) system. A region of the gene encoding the RNA-dependent RNA polymerase was amplified by using VSV-specific primers in the presence of two serotype-specific fluorescent probes. By incorporating...... probes. The limits of detection ware found to be less than 10 50% tissue culture infective doses/ml for both serotypes. The diagnostic value of the new method was tested with clinical materials from experimentally infected pigs, and it is concluded that the method is a powerful tool for the rapid...

  8. Diffusion-weighted MR imaging in postoperative follow-up: Reliability for detection of recurrent cholesteatoma

    Energy Technology Data Exchange (ETDEWEB)

    Cimsit, Nuri Cagatay [Marmara University Hospital, Department of Radiology, Istanbul (Turkey); Engin Sitesi Peker Sokak No:1 D:13, 34330 Levent, Istanbul (Turkey)], E-mail: cagataycimsit@gmail.com; Cimsit, Canan [Goztepe Education and Research Hospital, Department of Radiology, Istanbul (Turkey); Istanbul Goztepe Egitim ve Arastirma Hastanesi, Radyoloji Klinigi, Goztepe, Istanbul (Turkey)], E-mail: ccimsit@ttmail.com; Baysal, Begumhan [Goztepe Education and Research Hospital, Department of Radiology, Istanbul (Turkey); Istanbul Goztepe Egitim ve Arastirma Hastanesi, Radyoloji Klinigi, Goztepe, Istanbul (Turkey)], E-mail: begumbaysal@yahoo.com; Ruhi, Ilteris Cagatay [Goztepe Education and Research Hospital, Department of ENT, Istanbul (Turkey); Istanbul Goztepe Egitim ve Arastirma Hastanesi, KBB Klinigi, Goztepe, Istanbul (Turkey)], E-mail: cruhi@yahoo.com; Ozbilgen, Suha [Goztepe Education and Research Hospital, Department of ENT, Istanbul (Turkey); Istanbul Goztepe Egitim ve Arastirma Hastanesi, KBB Klinigi, Goztepe, Istanbul (Turkey)], E-mail: sozbilgen@yahoo.com; Aksoy, Elif Ayanoglu [Acibadem Bakirkoy Hospital, Department of ENT, Istanbul (Turkey); Acibadem Hastanesi, KBB Boeluemue, Bakirkoey, Istanbul (Turkey)], E-mail: elifayanoglu@yahoo.com

    2010-04-15

    Introduction: Cholesteatoma is a progressively growing process that destroy the neighboring bony structures and treatment is surgical removal. Follow-up is important in the postoperative period, since further surgery is necessary if recurrence is present, but not if granulation tissue is detected. This study evaluates if diffusion-weighted MR imaging alone can be a reliable alternative to CT, without use of contrast agent for follow-up of postoperative patients in detecting recurrent cholesteatoma. Materials and methods: 26 consecutive patients with mastoidectomy reporting for routine follow-up CT after mastoidectomy were included in the study, if there was loss of middle ear aeration on CT examination. MR images were evaluated for loss of aeration and signal intensity changes on diffusion-weighted sequences. Surgical results were compared with imaging findings. Results: Interpretation of MR images were parallel with the loss of aeration detected on CT for all 26 patients. Of the 26 patients examined, 14 were not evaluated as recurrent cholesteatoma and verified with surgery (NPV: 100%). Twelve patients were diagnosed as recurrent cholesteatoma and 11 were surgically diagnosed as recurrent cholesteatoma (PPV: 91.7%). Four of these 11 patients had loss of aeration size greater than the high signal intensity area on DWI, which were surgically confirmed as granulation tissue or fibrosis accompanying recurrent cholesteatoma. Conclusion: Diffusion-weighted MR for suspected recurrent cholesteatoma is a valuable tool to cut costs and prevent unnecessary second-look surgeries. It has the potential to become the MR sequence of choice to differentiate recurrent cholesteatoma from other causes of loss of aeration in patients with mastoidectomy.

  9. Dual-labeled time-resolved fluoroimmunoassay for simultaneous detection of clothianidin and diniconazole in agricultural samples.

    Science.gov (United States)

    Sheng, Enze; Shi, Haiyan; Zhou, Liangliang; Hua, Xiude; Feng, Lu; Yu, Tong; Wang, Minghua

    2016-02-01

    Europium (Eu(3+)) and samarium (Sm(3+)) were used as fluorescent labels to develop a highly sensitive dual-labeled time-resolved fluoroimmunoassay (TRFIA) for detect clothianidin and diniconazole in food samples. Under the optimized assay conditions, 50% inhibition concentration (IC50) and the limit of detection (LOD, IC10) of clothianidin were 5.08 and 0.021 μg/L, and 13.14 and 0.029 μg/L for diniconazole. The cross-reactivities (CRs) were negligible except dinotefuran (9.4%) and uniconazole (4.28%). The recoveries of clothianidin and diniconazole ranged from 79.3% to 108.7% in food samples. The results of TRFIA for the authentic samples were validated by gas chromatography (GC) analyses, and a satisfactory correlations were obtained. These results indicated that the method was an alternative tool for simultaneous detection of clothianidin and diniconazole in food samples. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. A novel multiplex PCR assay for simultaneous detection of nine clinically significant bacterial pathogens associated with bovine mastitis.

    Science.gov (United States)

    Ashraf, Aqeela; Imran, Muhammad; Yaqub, Tahir; Tayyab, Muhammad; Shehzad, Wasim; Thomson, Peter C

    2017-06-01

    For rapid and simultaneous detection of nine bovine mastitic pathogens, a sensitive and specific multiplex PCR assay was developed. The assay was standardized using reference strains and validated on mastitic milk cultures which were identified to species level based on 16S rRNA sequencing. Multiplex PCR assay also efficiently detected the target bacterial strains directly from milk. The detection limit of the assay was up to 50 pg for DNA isolated from pure cultures and 10 4  CFU/ml for spiked milk samples. As estimated by latent class analysis, the assay was sensitive up to 88% and specific up to 98% for targeted mastitic pathogens, compared with the bacterial culture method and the 16S rRNA sequence analysis. This novel molecular assay could be useful for monitoring and maintaining the bovine udder health, ensuring the bacteriological safety of milk, and conducting epidemiological studies. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Facile synthesis of graphene hybrid tube-like structure for simultaneous detection of ascorbic acid, dopamine, uric acid and tryptophan

    Energy Technology Data Exchange (ETDEWEB)

    Zhang Wen [Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Chai Yaqin, E-mail: yqchai@swu.edu.cn [Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Yuan Ruo, E-mail: yuanruo@swu.edu.cn [Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Chen Shihong; Han Jing; Yuan Dehua [Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China)

    2012-12-05

    Graphical abstract: A tube-like structure of graphene hybrid (GS-PTCA) was synthesized via {pi}-{pi} stacking interaction, and was used as modifier to fabricate electrode for simultaneous detection of ascorbic acid (AA), dopamine (DA), uric acid (UA) and tryptophan (Trp). SEM images of GS, PTCA and GS-PTCA were presented. Under the synergistic effects between GS and PTCA, the modified electrode displayed high catalytic activity and selectivity toward the oxidation of AA, DA, UA, and Trp. Highlights: Black-Right-Pointing-Pointer A simple strategy for simultaneous detection of AA, DA, UA and Trp has been constructed. Black-Right-Pointing-Pointer The tube-like structure of graphene hybrid (GS-PTCA) was synthesized. Black-Right-Pointing-Pointer The GS-PTCA provided a selective interface for discrimination of AA, DA, UA and Trp. - Abstract: In the present work, a tube-like structure of graphene hybrid as modifier to fabricate electrode for simultaneous detection of ascorbic acid (AA), dopamine (DA), uric acid (UA) and tryptophan (Trp) was reported. The hybrid was synthesized by a simple method based on graphene sheets (GS) and 3,4,9,10-perylenetetracarboxylic acid (PTCA) via {pi}-{pi} stacking interaction under ultrasonic condition. The combination of GS and PTCA could effectively improve the dispersion of GS, owing to PTCA with the carboxylic-functionalized interface. Comparing with pure GS or PTCA modified electrode, GS-PTCA displayed high catalytic activity and selectivity toward the oxidation of AA, DA, UA, and Trp. Moreover, cyclic voltammetry, different pulse voltammetry and scanning electron microscopy were employed to characterize the sensors. The experiment results showed that the linear response range for simultaneous detection of AA, DA, UA, and Trp were 20-420 {mu}M, 0.40-374 {mu}M, 4-544 {mu}M and 0.40-138 {mu}M, respectively, and the detection limits were 5.60 {mu}M, 0.13 {mu}M, 0.92 {mu}M and 0.06 {mu}M (S/N = 3). Importantly, the proposed method offers

  12. Cantilever-based micro-particle filter with simultaneous single particle detection

    DEFF Research Database (Denmark)

    Noeth, Nadine-Nicole; Keller, Stephan Sylvest; Boisen, Anja

    2011-01-01

    Currently, separation of whole blood samples on lab-on-a-chip systems is achieved via filters followed by analysis of the filtered matter such as counting of blood cells. Here, a micro-chip based on cantilever technology is developed, which enables simultaneous filtration and counting of micro-particles...... from a liquid. A hole-array is integrated into a micro-cantilever, which is inserted into a microfluidic channel perpendicular to the flow. A metal pad at the apex of the cantilever enables an optical read-out of the deflection of the cantilever. When a micro-particle is too large to pass a hole...

  13. A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages

    DEFF Research Database (Denmark)

    Muhammed, Musemma Kedir; Krych, Lukasz; Nielsen, Dennis Sandris

    2017-01-01

    simultaneous quantitative detection of Lc. lactis 936 (now SK1virus), P335, c2 (now C2virus) and Leuconostoc phage groups. Component assays are designed to have high efficiencies and nearly the same dynamic detection ranges, i.e., from 1.1 x 105 to 1.1 x 101 phage genomes per reaction, which corresponds to 9 x......Simultaneous quantitative detection of Lactococcus (Lc.) lactis and Leuconostoc species bacteriophages (phages) has not been reported in dairies using undefined mixed-strain DL-starters, probably due to the lack of applicable methods. We optimized a high-throughput qPCR system that allows...... 107 to 9 x 103 phage particles mL-1 without any additional up-concentrating steps. The amplification efficiencies of the corresponding assays were 100.1±2.6, 98.7±2.3, 101.0±2.3 and 96.2±6.2. The qPCR system was tested on samples obtained from a dairy plant that employed traditional mother...

  14. Development of a screening fluorescence polarization immunoassay for the simultaneous detection of fumonisins B₁ and B₂ in maize.

    Science.gov (United States)

    Li, Chenglong; Mi, Tiejun; Conti, Gea Oliveri; Yu, Qing; Wen, Kai; Shen, Jianzhong; Ferrante, Margherita; Wang, Zhanhui

    2015-05-27

    This paper reports the development of a screening fluorescence polarization immunoassay (FPIA) for the simultaneous detection of fumonisins B1 (FB1) and B2 (FB2) in maize. Three FB1 tracers including FB1-fluorescein isothiocyanate isomer I (FB1-FITC), FB1-5-([4,6-dichlorotriazine-2-yl]amino)-fluorescein (FB1-5-DTAF), and FB1-Texas Red-X succinimidyl ester (FB1-TRX) were synthesized and studied to select appropriate tracer-antibody pairs using seven previously produced monoclonal antibodies (mAbs). An FPIA employing the pair of FB1-FITC and mAb 4B9 showing 98.9% cross-reactivity (CR) toward FB2 was used to simultaneously detect FB1 and FB2. Maize flour samples were extracted with methanol/water (2:3, v/v). After optimization, the FPIA revealed a limit of detection (LOD) of 157.4 μg/kg for FB1 and an LOD of 290.6 μg/kg for FB2, respectively. Recoveries were measured for spiked samples of FB1 or FB2 separately, ranging from 84.7 to 93.6%, with a coefficient of variation (CV) of fumonisins in maize.

  15. A novel colorimetric sensor based on BODIPY-coumarin dye for simultaneous detection of cyanide and fluoride

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Yanhua, E-mail: hpyyh@aliyun.com [Institute for Interdisciplinary Research, Jianghan University, Wuhan 430056 (China); Shu, Tingting; Fu, Cheng; Yu, Bingjie; Zhang, Dongdong; Luo, Huixiu; Chen, Junjie [Institute for Interdisciplinary Research, Jianghan University, Wuhan 430056 (China); Dong, Changzhi [Institute for Interdisciplinary Research, Jianghan University, Wuhan 430056 (China); University Paris Diderot, Sorbonne Paris Cité, ITODYS, UMR CNRS 7086, 15 rue J-A de Baïf, 75205 Paris Cedex 13 (France)

    2017-06-15

    A novel colorimetric and fluorescent sensor 6 for fluoride and cyanide was developed based on BODIPY-coumarin platform and its anions sensing properties were investigated in the mixture of acetonitrile and Tris–HCl buffer (v/v = 95:5, pH = 7.5). Probe 6 could simultaneously detect F{sup –} and CN{sup –} through colorimetric method over the other competitive anions, such as Cl{sup –}, Br{sup –}, I{sup –}, NO{sub 3}{sup –}, ClO{sub 4}{sup –}, HSO{sub 4}{sup –}, S{sup 2–} and H{sub 2}PO{sub 4}{sup –}. It exhibited a distinct color change from red to green upon addition of F{sup –} through deprotection of tert-butyldiphenylsilyl group of coumarin. Moreover, it displayed an obvious color change from red to yellow through deprotection process firstly, then with a nucleophilic displacement mechanism. Therefore, the sensor 6 provides a novel method to simultaneously detect F{sup −} and CN{sup −} with different color change in the same solvent environment. The detection limit of sensor 6 toward F{sup –} and CN{sup –} ion was determined to be 0.43 μM and 1.9 μM respectively,.

  16. Simultaneous detection of xenon and krypton in equine plasma by gas chromatography-tandem mass spectrometry for doping control.

    Science.gov (United States)

    Kwok, Wai Him; Choi, Timmy L S; So, Pui-Kin; Yao, Zhong-Ping; Wan, Terence S M

    2017-02-01

    Xenon can activate the hypoxia-inducible factors (HIFs). As such, it has been allegedly used in human sports for increasing erythropoiesis. Krypton, another noble gas with reported narcosis effect, can also be expected to be a potential and less expensive erythropoiesis stimulating agent. This has raised concern about the misuse of noble gases as doping agents in equine sports. The aim of the present study is to establish a method for the simultaneous detection of xenon and krypton in equine plasma for the purpose of doping control. Xenon- or krypton-fortified equine plasma samples were prepared according to reported protocols. The target noble gases were simultaneously detected by gas chromatography-triple quadrupole mass spectrometry using headspace injection. Three xenon isotopes at m/z 129, 131, and 132, and four krypton isotopes at m/z 82, 83, 84, and 86 were targeted in selected reaction monitoring mode (with the precursor ions and product ions at identical mass settings), allowing unambiguous identification of the target analytes. Limits of detection for xenon and krypton were about 19 pmol/mL and 98 pmol/mL, respectively. Precision for both analytes was less than 15%. The method has good specificity as background analyte signals were not observed in negative equine plasma samples (n = 73). Loss of analytes under different storage temperatures has also been evaluated. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  17. Simultaneous Detection of Both Single Nucleotide Variations and Copy Number Alterations by Next-Generation Sequencing in Gorlin Syndrome.

    Directory of Open Access Journals (Sweden)

    Kei-ichi Morita

    Full Text Available Gorlin syndrome (GS is an autosomal dominant disorder that predisposes affected individuals to developmental defects and tumorigenesis, and caused mainly by heterozygous germline PTCH1 mutations. Despite exhaustive analysis, PTCH1 mutations are often unidentifiable in some patients; the failure to detect mutations is presumably because of mutations occurred in other causative genes or outside of analyzed regions of PTCH1, or copy number alterations (CNAs. In this study, we subjected a cohort of GS-affected individuals from six unrelated families to next-generation sequencing (NGS analysis for the combined screening of causative alterations in Hedgehog signaling pathway-related genes. Specific single nucleotide variations (SNVs of PTCH1 causing inferred amino acid changes were identified in four families (seven affected individuals, whereas CNAs within or around PTCH1 were found in two families in whom possible causative SNVs were not detected. Through a targeted resequencing of all coding exons, as well as simultaneous evaluation of copy number status using the alignment map files obtained via NGS, we found that GS phenotypes could be explained by PTCH1 mutations or deletions in all affected patients. Because it is advisable to evaluate CNAs of candidate causative genes in point mutation-negative cases, NGS methodology appears to be useful for improving molecular diagnosis through the simultaneous detection of both SNVs and CNAs in the targeted genes/regions.

  18. Simultaneous Detection of Both Single Nucleotide Variations and Copy Number Alterations by Next-Generation Sequencing in Gorlin Syndrome.

    Science.gov (United States)

    Morita, Kei-ichi; Naruto, Takuya; Tanimoto, Kousuke; Yasukawa, Chisato; Oikawa, Yu; Masuda, Kiyoshi; Imoto, Issei; Inazawa, Johji; Omura, Ken; Harada, Hiroyuki

    2015-01-01

    Gorlin syndrome (GS) is an autosomal dominant disorder that predisposes affected individuals to developmental defects and tumorigenesis, and caused mainly by heterozygous germline PTCH1 mutations. Despite exhaustive analysis, PTCH1 mutations are often unidentifiable in some patients; the failure to detect mutations is presumably because of mutations occurred in other causative genes or outside of analyzed regions of PTCH1, or copy number alterations (CNAs). In this study, we subjected a cohort of GS-affected individuals from six unrelated families to next-generation sequencing (NGS) analysis for the combined screening of causative alterations in Hedgehog signaling pathway-related genes. Specific single nucleotide variations (SNVs) of PTCH1 causing inferred amino acid changes were identified in four families (seven affected individuals), whereas CNAs within or around PTCH1 were found in two families in whom possible causative SNVs were not detected. Through a targeted resequencing of all coding exons, as well as simultaneous evaluation of copy number status using the alignment map files obtained via NGS, we found that GS phenotypes could be explained by PTCH1 mutations or deletions in all affected patients. Because it is advisable to evaluate CNAs of candidate causative genes in point mutation-negative cases, NGS methodology appears to be useful for improving molecular diagnosis through the simultaneous detection of both SNVs and CNAs in the targeted genes/regions.

  19. Improvement of the qualitative and quantitative detection of simultaneously present fluorescent tracers by systematic sample treatment

    International Nuclear Information System (INIS)

    Behrens, H.

    1982-01-01

    The selective instrumental detection of individual fluorescent tracers in mixtures containing further fluorescent dyes is limited by spectral interferences. Therefore additional separations or other suitable procedures have to be included into the analytic technique. With the method described below, the respective tracer to be detected remains with its initial concentration in the sample and is analysed under the appropriate conditions, whereas the interfering tracers are separated or suppressed. The techniques applied for this base on the facts that 1) the fluorescence intensity of the tracers varies differently when the pH-value changes; 2) the tracers show different absorption behaviour and 3) they provide different degrees of light sensitivity. The procedures permit for example to detect uranin when eosin is present in a higher concentration or to detect eosin when amidorhodamin G is present. (orig.) [de

  20. Colloidal gold-McAb probe-based rapid immunoassay strip for simultaneous detection of fumonisins in maize.

    Science.gov (United States)

    Yao, Jingjing; Sun, Yaning; Li, Qingmei; Wang, Fangyu; Teng, Man; Yang, Yanyan; Deng, Ruiguang; Hu, Xiaofei

    2017-05-01

    Fumonisins are a kind of toxic and carcinogenic mycotoxin. A rapid immunochromatographic test strip has been developed for simultaneous detection of fumonisin B 1 , B 2 and B 3 (FB 1 , FB 2 and FB 3 ) in maize based on colloidal gold-labelled monoclonal antibody (McAb) against FB 1 probe. The anti-FB 1 McAb (2E11-H3) was produced through immunisation and cell fusion, and identified as high affinity, specificity and sensitivity. The cross-reaction ratios with fumonisin B 2 and B 3 were accordingly 385% and 72.4%, while none with other analogues. The colloid gold-labelled anti-FB 1 McAb probe was successfully prepared and used for establishing the immunochromatographic strip. The test strip showed high sensitivity and specificity, the IC 50 for FB 1 was 58.08 ng mL -1 , LOD was 11.24 ng mL -1 , calculated from standard curve. Moreover, the test strip exhibited high cross-reactivity with FB 2 and FB 3 , and could be applied to the simultaneous detection of FBs (FB 1 :FB 2 :FB 3 = 12:4:1) in maize sample with high accuracy and precision. The average recoveries of FBs in maize ranged from 90.42% to 95.29%, and CVs were 1.25-3.77%. The results of the test strip for FBs samples showed good correlation with high-performance liquid chromatography analysis. The immunochromatographic test strip could be employed in the rapid simultaneous detection of FB 1 , FB 2 and FB 3 in maize samples on-site. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  1. Simultaneous differential detection of Chlamydophila abortus, Chlamydophila pecorum and Coxiella burnetii from aborted ruminant's clinical samples using multiplex PCR

    Directory of Open Access Journals (Sweden)

    Rodolakis Annie

    2009-07-01

    Full Text Available Abstract Background Chlamydiosis and Q fever, two zoonosis, are important causes of ruminants' abortion around the world. They are caused respectively by strictly intracellular and Gram negative bacterium Chlamydophila abortus (Cp. abortus and Coxiella burnetii (C. burnetii. Chlamydophila pecorum (Cp. pecorum is commonly isolated from the digestive tract of clinically inconspicuous ruminants but the abortive and zoonotic impact of this bacterium is still unknown because Cp. pecorum is rarely suspected in abortion cases of small ruminants. We have developed a multiplex PCR (m-PCR for rapid simultaneous differential detection of Cp. abortus, Cp. pecorum and C. burnetii in clinical samples taken from infected animals. Results Specific PCR primers were designed and a sensitive and specific m-PCR was developed to detect simultaneously, in one tube reaction, three specific fragments of 821, 526 and 687-bp long for Cp. abortus, Cp. pecorum and C. burnetii respectively. This m-PCR assay was performed on 253 clinical samples taken from infected ruminant's flocks that have showed problems of abortion diseases. Thus, 67 samples were infected by either one of the three pathogens: 16 (13 vaginal swabs and 3 placentas were positive for Cp. abortus, 2 were positive for Cp. pecorum (1 vaginal swab and 1 placenta and 49 samples (33 vaginal swabs, 11 raw milks, 4 faeces and 1 placenta were positive for C. burnetii. Two vaginal swabs were m-PCR positive of both Cp. abortus and C. burnetii and none of the tested samples was shown to be infected simultaneously with the three pathogens. Conclusion We have successfully developed a rapid multiplex PCR that can detect and differentiate Cp. abortus, Cp. pecorum and C. burnetii; with a good sensitivity and specificity. The diagnosis of chlamydiosis and Q fever may be greatly simplified and performed at low cost. In addition, the improvement in diagnostic techniques will enhance our knowledge regarding the prevalence and

  2. Simultaneous and rapid determination of deoxynivalenol and its acetylate derivatives in wheat flour and rice by ultra high performance liquid chromatography with photo diode array detection.

    Science.gov (United States)

    Xu, Jiao-Jiao; Zhou, Jian; Huang, Bai-Fen; Cai, Zeng-Xuan; Xu, Xiao-Min; Ren, Yi-Ping

    2016-06-01

    A simple and reliable method of ultra high performance liquid chromatography coupled with photo-diode array detection has been proposed for the simultaneous determination of deoxynivalenol and its acetylated derivatives in wheat flour and rice, especially focusing on the optimization of sample extraction, cleanup, and chromatographic separation conditions. Sample pretreatment consisted of a first step using a quick, easy, cheap, effective, rugged, and safe based extraction procedure and a subsequent cleanup step based on solid-phase extraction. The method was extensively validated in wheat flour and rice, obtaining satisfactory analytical performance with good linearity (R(2) ≥ 0.999), acceptable recoveries (80.0-104.4%), and repeatability (RSDs 1.3-10.7%). The limits of detection (21.7-57.4 μg/kg) and quantitation (72.3-191.4 μg/kg) for deoxynivalenols were lower than those usually permitted by various countries' legislation in these food matrices. The method was applied to 34 wheat and rice samples. The results were further compared with results of ultra high performance liquid chromatography with electrospray ionization tandem mass spectrometry. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Simultaneous determination of 11 fluorescent whitening agents in food-contact paper and board by ion-pairing high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Jiang, Dingguo; Chen, Lisong; Fu, Wusheng; Qiu, Hanquan

    2015-02-01

    4,4'-Diaminostilbene-2,2'-disulfonic acid based fluorescent whitening agents (DSD-FWAs) are prohibited in food-contact paper and board in many countries. In this work, a reliable high-performance liquid chromatography method was developed for the simultaneous determination of 11 common DSD-FWAs in paper material. Sample preparation and extraction as well as chromatographic separation of multicomponent DSD-FWAs were successfully optimized. DSD-FWAs in prepared samples were ultrasonically extracted with acetonitrile/water/triethylamine (40:60:1, v/v/v), separated on the C(18) column with the mobile phase containing tetrabutylammonium bromide, and then detected by a fluorescence detector. The limits of detection were 0.12-0.24 mg/kg, and the calibration curves showed the linear correlation (R(2) ≥ 0.9994) within the range of 8.0-100 ng/mL, which was equivalent to the range of 0.80-10 mg/kg in the sample. The average recoveries and the RSDs were 81-106% and 2-9% at two fortification levels (1.0 and 5.0 mg/kg) in paper bowls, respectively. The successful determination of 11 DSD-FWAs in food-contact paper and board obtained from local markets indicated that the newly developed method was rapid, accurate, and highly selective. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Simultaneous detection and quantification of Phytophthora nicotianae and P. cactorum, and distribution analyses in strawberry greenhouses by duplex real-time PCR.

    Science.gov (United States)

    Li, Mingzhu; Inada, Minoru; Watanabe, Hideki; Suga, Haruhisa; Kageyama, Koji

    2013-01-01

    Phytophthora nicotianae and P. cactorum cause Phytophthora rot of strawberry. A duplex real-time PCR technique for simultaneous detection and quantification of the two pathogens was developed. Species-specific primers for P. nicotianae and P. cactorum were designed based on the internal transcribed spacer regions (ITS) of rDNA and the ras-related protein gene Ypt1, respectively. TaqMan probes were labeled with FAM for P. nicotianae and HEX for P. cactorum. Specificities were demonstrated using 52 isolates, including various soil-borne pathogens. Sensitivities for P. nicotianae and P. cactorum DNAs were 10 fg and 1 pg, respectively. The technique was applied to naturally infested soil and root samples; the two pathogens were detected and the target DNA concentrations were quantified. Significant correlations of DNA quantities in roots and the surrounding soils were found. The minimum soil DNA concentration predicting the development of disease symptoms was estimated as 20 pg (g soil)(-1). In three strawberry greenhouses examined, the target DNA concentrations ranged from 1 to 1,655 pg (g soil)(-1) for P. nicotianae and from 13 to 233 pg (g soil)(-1) for P. cactorum. The method proved fast and reliable, and provides a useful tool to monitor P. nicotianae and P. cactorum in plants or soils.

  5. Aptamer- and nucleic acid enzyme-based systems for simultaneous detection of multiple analytes

    Science.gov (United States)

    Lu, Yi [Champaign, IL; Liu, Juewen [Albuquerque, NM

    2011-11-15

    The present invention provides aptamer- and nucleic acid enzyme-based systems for simultaneously determining the presence and optionally the concentration of multiple analytes in a sample. Methods of utilizing the system and kits that include the sensor components are also provided. The system includes a first reactive polynucleotide that reacts to a first analyte; a second reactive polynucleotide that reacts to a second analyte; a third polynucleotide; a fourth polynucleotide; a first particle, coupled to the third polynucleotide; a second particle, coupled to the fourth polynucleotide; and at least one quencher, for quenching emissions of the first and second quantum dots, coupled to the first and second reactive polynucleotides. The first particle includes a quantum dot having a first emission wavelength. The second particle includes a second quantum dot having a second emission wavelength different from the first emission wavelength. The third polynucleotide and the fourth polynucleotide are different.

  6. Simultaneous Power Deposition Detection of Two EC Beams with the BIS Analysis in Moving TCV Plasmas

    Science.gov (United States)

    Curchod, L.; Pochelon, A.; Decker, J.; Felici, F.; Goodman, T. P.; Moret, J.-M.; Paley, J. I.

    2009-11-01

    Modulation of power amplitude is a widespread to determine the radial absorption profile of externally launched power in fusion plasmas. There are many techniques to analyze the plasma response to such a modulation. The break-in-slope (BIS) analysis can draw an estimated power deposition profile for each power step up. In this paper, the BIS analysis is used to monitor the power deposition location of one or two EC power beams simultaneously in a non-stationary plasma being displaced vertically in the TCV tokamak vessel. Except from radial discrepancies, the results have high time resolution and compare well with simulations from the R2D2-C3PO-LUKE ray-tracing and Fokker-Planck code suite.

  7. Simultaneous ion detection in a mass spectrometer with variable mass dispersion

    International Nuclear Information System (INIS)

    Tuithof, H.H.

    1977-01-01

    This thesis mainly describes the ion-optics of a magnetic mass spectrometer system, especially applied to the projection of a significant part of the mass spectrum onto a flat ion-detector. The complete detector consists of a channeltron electron multiplier array with phosphor screen and a Vidicon-multichannel analyzer combination for simultaneous read-out. In order to optimise the spectral range projected onto the channelplate, by varying the mass dispersion and to rotate the oblique angle of the mass focal plane with respect to the detector surface, the sector magnet has been combined with electrostatic and magnetic quadrupole lenses. This detector will find wide application in the analysis of minute sample quantities, in the recording of extremely short ion events (large molecules) and at collision activation mass-spectrometry studies

  8. OCT4 and SOX2 are reliable markers in detecting stem cells in odontogenic lesions

    Directory of Open Access Journals (Sweden)

    Abhishek Banerjee

    2016-01-01

    Full Text Available Context (Background: Stem cells are a unique subpopulation of cells in the human body with a capacity to initiate differentiation into various cell lines. Tumor stem cells (TSCs are a unique subpopulation of cells that possess the ability to initiate a neoplasm and sustain self-renewal. Epithelial stem cell (ESC markers such as octamer-binding transcription factor 4 (OCT4 and sex-determining region Y (SRY-box 2 (SOX2 are capable of identifying these stem cells expressed during the early stages of tooth development. Aims: To detect the expression of the stem cell markers OCT4 and SOX2 in the normal odontogenic tissues and the odontogenic cysts and tumors. Materials and Methods: Paraffin sections of follicular tissue, radicular cyst, dentigerous cyst, odontogenic keratocyst, ameloblastoma, adenomatoid odontogenic tumor, and ameloblastic carcinoma were obtained from the archives. The sections were subjected to immunohistochemical assay by the use of mouse monoclonal antibodies to OCT4 and SOX2. Statistical Analysis: The results were evaluated by descriptive analysis. Results: The results show the presence of stem cells in the normal and lesional tissues with these stem cell identifying markers. SOX2 was found to be more consistent and reliable in the detection of stem cells. Conclusion: The stem cell expressions are maintained in the tumor transformation of tissue and probably suggest that there is no phenotypic change of stem cells in progression from normal embryonic state to its tumor component. The quantification and localization reveals interesting trends that indicate the probable role of the cells in the pathogenesis of the lesions.

  9. Reliability of magnetic resonance imaging for the detection of hypopituitarism in children with optic nerve hypoplasia.

    Science.gov (United States)

    Ramakrishnaiah, Raghu H; Shelton, Julie B; Glasier, Charles M; Phillips, Paul H

    2014-01-01

    It is essential to identify hypopituitarism in children with optic nerve hypoplasia (ONH) because they are at risk for developmental delay, seizures, or death. The purpose of this study is to determine the reliability of neurohypophyseal abnormalities on magnetic resonance imaging (MRI) for the detection of hypopituitarism in children with ONH. Cross-sectional study. One hundred one children with clinical ONH who underwent MRI of the brain and orbits and a detailed pediatric endocrinologic evaluation. Magnetic resonance imaging studies were performed on 1.5-Tesla scanners. The imaging protocol included sagittal T1-weighted images, axial fast fluid-attenuated inversion-recovery/T2-weighted images, and diffusion-weighted images of the brain. Orbital imaging included fat-saturated axial and coronal images and high-resolution axial T2-weighted images. The MRI studies were reviewed by 2 pediatric neuroradiologists for optic nerve hypoplasia, absent or ectopic posterior pituitary, absent pituitary infundibulum, absent septum pellucidum, migration anomalies, and hemispheric injury. Medical records were reviewed for clinical examination findings and endocrinologic status. All patients underwent a clinical evaluation by a pediatric endocrinologist and a standardized panel of serologic testing that included serum insulin-like growth factor-1, insulin-like growth factor binding protein-3, prolactin, cortisol, adrenocorticotropic hormone, thyroid-stimulating hormone, and free thyroxine levels. Radiologists were masked to patients' endocrinologic status and funduscopic findings. Sensitivity and specificity of MRI findings for the detection of hypopituitarism. Neurohypophyseal abnormalities, including absent pituitary infundibulum, ectopic posterior pituitary bright spot, and absent posterior pituitary bright spot, occurred in 33 children. Magnetic resonance imaging disclosed neurohypophyseal abnormalities in 27 of the 28 children with hypopituitarism (sensitivity, 96%). A

  10. Human reliability-based MC and A models for detecting insider theft

    International Nuclear Information System (INIS)

    Duran, Felicia Angelica; Wyss, Gregory Dane

    2010-01-01

    Material control and accounting (MC and A) safeguards operations that track and account for critical assets at nuclear facilities provide a key protection approach for defeating insider adversaries. These activities, however, have been difficult to characterize in ways that are compatible with the probabilistic path analysis methods that are used to systematically evaluate the effectiveness of a site's physical protection (security) system (PPS). MC and A activities have many similar characteristics to operator procedures performed in a nuclear power plant (NPP) to check for anomalous conditions. This work applies human reliability analysis (HRA) methods and models for human performance of NPP operations to develop detection probabilities for MC and A activities. This has enabled the development of an extended probabilistic path analysis methodology in which MC and A protections can be combined with traditional sensor data in the calculation of PPS effectiveness. The extended path analysis methodology provides an integrated evaluation of a safeguards and security system that addresses its effectiveness for attacks by both outside and inside adversaries.

  11. How often should we monitor for reliable detection of atrial fibrillation recurrence? Efficiency considerations and implications for study design.

    Directory of Open Access Journals (Sweden)

    Efstratios I Charitos

    Full Text Available Although atrial fibrillation (AF recurrence is unpredictable in terms of onset and duration, current intermittent rhythm monitoring (IRM diagnostic modalities are short-termed and discontinuous. The aim of the present study was to investigate the necessary IRM frequency required to reliably detect recurrence of various AF recurrence patterns.The rhythm histories of 647 patients (mean AF burden: 12 ± 22% of monitored time; 687 patient-years with implantable continuous monitoring devices were reconstructed and analyzed. With the use of computationally intensive simulation, we evaluated the necessary IRM frequency to reliably detect AF recurrence of various AF phenotypes using IRM of various durations.The IRM frequency required for reliable AF detection depends on the amount and temporal aggregation of the AF recurrence (p95% sensitivity of AF recurrence required higher IRM frequencies (>12 24-hour; >6 7-day; >4 14-day; >3 30-day IRM per year; p<0.0001 than currently recommended. Lower IRM frequencies will under-detect AF recurrence and introduce significant bias in the evaluation of therapeutic interventions. More frequent but of shorter duration, IRMs (24-hour are significantly more time effective (sensitivity per monitored time than a fewer number of longer IRM durations (p<0.0001.Reliable AF recurrence detection requires higher IRM frequencies than currently recommended. Current IRM frequency recommendations will fail to diagnose a significant proportion of patients. Shorter duration but more frequent IRM strategies are significantly more efficient than longer IRM durations.Unique identifier: NCT00806689.

  12. A method for detecting IBD regions simultaneously in multiple individuals--with applications to disease genetics

    DEFF Research Database (Denmark)

    Moltke, Ida; Albrechtsen, Anders; Hansen, Thomas V O

    2011-01-01

    genome containing disease-causing variants. However, IBD regions can be difficult to detect, especially in the common case where no pedigree information is available. In particular, all existing non-pedigree based methods can only infer IBD sharing between two individuals. Here, we present a new Markov...... Chain Monte Carlo method for detection of IBD regions, which does not rely on any pedigree information. It is based on a probabilistic model applicable to unphased SNP data. It can take inbreeding, allele frequencies, genotyping errors, and genomic distances into account. And most importantly, it can...

  13. Simultaneous detection and classification of breast masses in digital mammograms via a deep learning YOLO-based CAD system.

    Science.gov (United States)

    Al-Masni, Mohammed A; Al-Antari, Mugahed A; Park, Jeong-Min; Gi, Geon; Kim, Tae-Yeon; Rivera, Patricio; Valarezo, Edwin; Choi, Mun-Taek; Han, Seung-Moo; Kim, Tae-Seong

    2018-04-01

    Automatic detection and classification of the masses in mammograms are still a big challenge and play a crucial role to assist radiologists for accurate diagnosis. In this paper, we propose a novel Computer-Aided Diagnosis (CAD) system based on one of the regional deep learning techniques, a ROI-based Convolutional Neural Network (CNN) which is called You Only Look Once (YOLO). Although most previous studies only deal with classification of masses, our proposed YOLO-based CAD system can handle detection and classification simultaneously in one framework. The proposed CAD system contains four main stages: preprocessing of mammograms, feature extraction utilizing deep convolutional networks, mass detection with confidence, and finally mass classification using Fully Connected Neural Networks (FC-NNs). In this study, we utilized original 600 mammograms from Digital Database for Screening Mammography (DDSM) and their augmented mammograms of 2,400 with the information of the masses and their types in training and testing our CAD. The trained YOLO-based CAD system detects the masses and then classifies their types into benign or malignant. Our results with five-fold cross validation tests show that the proposed CAD system detects the mass location with an overall accuracy of 99.7%. The system also distinguishes between benign and malignant lesions with an overall accuracy of 97%. Our proposed system even works on some challenging breast cancer cases where the masses exist over the pectoral muscles or dense regions. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Gamma spectrometry analysis for simultaneous detection of 54Mn, 65Zn and 59Fe in aqueous solutions and plant tissues

    International Nuclear Information System (INIS)

    Montanheiro, M.N.S.

    1975-01-01

    A methodology to detect the activities of 54 Mn, 65 Zn and 59 Fe in the same sample, with a single channel spectrometer coupled to a scintilation detector of NaI(tl), 3'' x 3'', well type has been developed. Initially a selection of the energy channel was made based on the criteria of maximizing the signal-background ratio and consequently, the minimization of the variation coefficient. In the channels, a study of minimal detectable activities was conducted for each radioisotopes. Secondly, samples containing diferent combinations of these radioisotopes were prepared and their activities were calculated using simultaneous equations. As a mean of demonstrating the pratical utility of this methodology, an experiment was developed in which the roots, isolated from beam plants (Phaseolus vulgaris, L.) were examined to determine levels of ionic absorption interference among micronutrients (Mn, Zn and Fe)

  15. A sensor of a polyoxometalate and Au–Pd alloy for simultaneously detection of dopamine and ascorbic acid

    International Nuclear Information System (INIS)

    Zhou, Chunlei; Li, Shuang; Zhu, Wei; Pang, Haijun; Ma, Huiyuan

    2013-01-01

    A composite film based on K 8 P 2 W 16 V 2 O 62 ·18H 2 O decorated by Au–Pd nanoparticles was prepared by the layer-by-layer self-assembly method. This composite film exhibits enhanced electrocatalytic performance, repeatability and long-term stability for the simultaneous determination of dopamine and ascorbic acid at biological pH (pH 7.0). The proposed electrochemical sensing film for simultaneous detecting dopamine and ascorbic acid shows rather low detection limit of 8.3 × 10 −7 and 4.3 × 10 −7 M and a linear response range from 2.1 × 10 −6 to 2.06 × 10 −3 M and 1.2 × 10 −6 to 1.61 × 10 −3 M, as well as no interference from the common interfering species at an applied potential. -- Abstract: A novel composite film based on Dawson-type phosphovanadotungstate K 8 P 2 W 16 V 2 O 62 ·18H 2 O (P 2 W 16 V 2 ) decorated by Au–Pd alloy nanoparticles (Au–Pd) was fabricated on quartz, silicon and ITO using the layer-by-layer self-assembly method. The composite film was characterized by UV–vis spectroscopy, X-ray photoelectron spectra, atomic force microscopy, Scanning electronic microscope, cyclic voltammetry and differential pulse voltammetry. The composite film can be employed for sensitive and simultaneous determination of dopamine and ascorbic acid at biological pH (pH 7.0). Linear analytical curves were obtained in the ranges from 2.1 × 10 −6 to 2.06 × 10 −3 M and 1.2 × 10 −6 to 1.61 × 10 −3 M for dopamine and ascorbic acid by DPV methods, respectively. The low detection limit for dopamine and ascorbic acid were 8.3 × 10 −7 and 4.3 × 10 −7 M, as well as no interference was observed from the common interfering species such as glucose, uric acid, L-cysteine, CH 3 OH, CH 3 CH 2 OH and H 2 O 2 . The composite film was used for dopamine and ascorbic acid determinations in real samples with satisfactory results. With high sensitivity and selectivity, the proposed electrochemical sensor would provide a simple method for

  16. Simultaneous detection of peanut and hazelnut allergens in food matrices using multiplex PCR method

    Directory of Open Access Journals (Sweden)

    Eva Renčová

    2014-01-01

    Full Text Available Multiplex PCR analysis for the detection of two targeting segments of genes coding major food protein allergens as peanut (Arachis hypogaea Ara h 1 gene and hazelnut (Corylus avellana Cor a 1 gene was developed. Two sets of primers were designed and tested to their specificity on a broad range of ingredients. The identity of amplicons (Ara h 1- 180 bp, Cor a 1 – 258 bp by sequencing and alignment of sequences with sequences deposited in Genbank was confirmed. When testing the specificity of designed primer pairs on a spectrum of food ingredients, no cross reactions were detected. A potential inhibition of PCR reaction was eliminated using the universal plant primers of chloroplast gene 124 bp for the plant matrices confirmation. The intrinsic detection limit was 10 pg·ml-1 and the practical detection limit was 0.001% w/w (10 mg·kg-1 for both peanuts and hazelnuts. The method was applied to the investigation of 60 commercial food samples. The developed multiplex PCR method is cheap, specific and sensitive enough and can be used as a simple, one day procedure for the checking of undeclared peanut and hazelnut major allergens in food.

  17. Simultaneous Event-Triggered Fault Detection and Estimation for Stochastic Systems Subject to Deception Attacks.

    Science.gov (United States)

    Li, Yunji; Wu, QingE; Peng, Li

    2018-01-23

    In this paper, a synthesized design of fault-detection filter and fault estimator is considered for a class of discrete-time stochastic systems in the framework of event-triggered transmission scheme subject to unknown disturbances and deception attacks. A random variable obeying the Bernoulli distribution is employed to characterize the phenomena of the randomly occurring deception attacks. To achieve a fault-detection residual is only sensitive to faults while robust to disturbances, a coordinate transformation approach is exploited. This approach can transform the considered system into two subsystems and the unknown disturbances are removed from one of the subsystems. The gain of fault-detection filter is derived by minimizing an upper bound of filter error covariance. Meanwhile, system faults can be reconstructed by the remote fault estimator. An recursive approach is developed to obtain fault estimator gains as well as guarantee the fault estimator performance. Furthermore, the corresponding event-triggered sensor data transmission scheme is also presented for improving working-life of the wireless sensor node when measurement information are aperiodically transmitted. Finally, a scaled version of an industrial system consisting of local PC, remote estimator and wireless sensor node is used to experimentally evaluate the proposed theoretical results. In particular, a novel fault-alarming strategy is proposed so that the real-time capacity of fault-detection is guaranteed when the event condition is triggered.

  18. Vapor permeation-stepwise injection simultaneous determination of methanol and ethanol in biodiesel with voltammetric detection.

    Science.gov (United States)

    Shishov, Andrey; Penkova, Anastasia; Zabrodin, Andrey; Nikolaev, Konstantin; Dmitrenko, Maria; Ermakov, Sergey; Bulatov, Andrey

    2016-02-01

    A novel vapor permeation-stepwise injection (VP-SWI) method for the determination of methanol and ethanol in biodiesel samples is discussed. In the current study, stepwise injection analysis was successfully combined with voltammetric detection and vapor permeation. This method is based on the separation of methanol and ethanol from a sample using a vapor permeation module (VPM) with a selective polymer membrane based on poly(phenylene isophtalamide) (PA) containing high amounts of a residual solvent. After the evaporation into the headspace of the VPM, methanol and ethanol were transported, by gas bubbling, through a PA membrane to a mixing chamber equipped with a voltammetric detector. Ethanol was selectively detected at +0.19 V, and both compounds were detected at +1.20 V. Current subtractions (using a correction factor) were used for the selective determination of methanol. A linear range between 0.05 and 0.5% (m/m) was established for each analyte. The limits of detection were estimated at 0.02% (m/m) for ethanol and methanol. The sample throughput was 5 samples h(-1). The method was successfully applied to the analysis of biodiesel samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Simultaneous detection of transgenic DNA by surface plasmon resonance imaging with potential application to gene doping detection.

    Science.gov (United States)

    Scarano, Simona; Ermini, Maria Laura; Spiriti, Maria Michela; Mascini, Marco; Bogani, Patrizia; Minunni, Maria

    2011-08-15

    Surface plasmon resonance imaging (SPRi) was used as the transduction principle for the development of optical-based sensing for transgenes detection in human cell lines. The objective was to develop a multianalyte, label-free, and real-time approach for DNA sequences that are identified as markers of transgenosis events. The strategy exploits SPRi sensing to detect the transgenic event by targeting selected marker sequences, which are present on shuttle vector backbone used to carry out the transfection of human embryonic kidney (HEK) cell lines. Here, we identified DNA sequences belonging to the Cytomegalovirus promoter and the Enhanced Green Fluorescent Protein gene. System development is discussed in terms of probe efficiency and influence of secondary structures on biorecognition reaction on sensor; moreover, optimization of PCR samples pretreatment was carried out to allow hybridization on biosensor, together with an approach to increase SPRi signals by in situ mass enhancement. Real-time PCR was also employed as reference technique for marker sequences detection on human HEK cells. We can foresee that the developed system may have potential applications in the field of antidoping research focused on the so-called gene doping.

  20. [Western Blot diagnostic yield for simultaneous antibody-detection in patients with human cysticercosis, hydatidosis, and human fascioliasis].

    Science.gov (United States)

    Davelois, Kelly; Escalante, Hermes; Jara, César

    2016-01-01

    . To determine the diagnostic yield using western blotting to simultaneously detect antibodies in patients with human cysticercosis, hydatidosis, and human fascioliasis. Materials and methods . Cross-sectional study of diagnostic yield assessment. Excretory/secretory antigens were obtained from Taenia solium larvae, Echinococcus granulosus cysts, and the adult flukes of Fasciola hepática, which were then separated using the polyacrylamide gel electrophoresis technique, transferred, and attached to a nitrocellulose membrane to be probed with sera from the patient infected with the three parasites. The sensitivity of the technique was assessed using 300 individual serum samples, 60 pools of two parasites, and 20 pools of three parasites with 75 sera from patients with other parasites, 10 from patients with other diseases, and 15 from patients without parasites. Results . The technique revealed 13 glycoproteins (GP): GP 35, 31, 24, 23, 18, 17, 14, and 13 kDa for cysticercosis; GP 8, 16, and 21 kDa for hydatidosis; and GP 17 and 23 kDa for fascioliasis. The test detected the presence of antibodies with a sensitivity of 96% (95% confidence interval [CI] = 94.62-98.54%) in the detection of one or the thirteen bands, a specificity of 100% (95% CI = 99.50-100.00%); individually, there was a sensitivity for cysticercosis of 97% (95% CI = 93.16-100.00%), for hydatidosis of 94% (95% CI = 88.85-99.15%) and for fascioliasis of 96% (95% CI = 91.66-100.00%). Conclusions . Western blotting is effective in the simultaneous detection of antibodies in patients with human cysticercosis, hydatidosis, and fascioliasis, and it can be used as a diagnostic test to either rule out or confirm the presence of antibodies in endemic areas.

  1. What’s hampering measurement invariance: Detecting non-invariant items using clusterwise simultaneous component analysis

    Directory of Open Access Journals (Sweden)

    Kim eDe Roover

    2014-06-01

    Full Text Available The issue of measurement invariance is ubiquitous in the behavioral sciences nowadays as more and more studies yield multivariate multigroup data. When measurement invariance cannot be established across groups, this is often due to different loadings on only a few items. Within the multigroup CFA framework, methods have been proposed to trace such non-invariant items, but these methods have some disadvantages in that they require researchers to run a multitude of analyses and in that they imply assumptions that are often questionable. In this paper, we propose an alternative strategy which builds on clusterwise simultaneous component analysis (SCA. Clusterwise SCA, being an exploratory technique, assigns the groups under study to a few clusters based on differences and similarities in the covariance matrices, and thus based on the component structure of the items. Non-invariant items can then be traced by comparing the cluster-specific component loadings via congruence coefficients, which is far more parsimonious than comparing the component structure of all separate groups. In this paper we present a heuristic for this procedure. Afterwards, one can return to the multigroup CFA framework and check whether removing the non-invariant items or removing some of the equality restrictions for these items, yields satisfactory invariance test results. An empirical application concerning cross-cultural emotion data is used to demonstrate that this novel approach is useful and can co-exist with the traditional CFA approaches.

  2. Combining optical trapping in a microfluidic channel with simultaneous micro-Raman spectroscopy and motion detection

    Science.gov (United States)

    Lawton, Penelope F.; Saunter, Christopher D.; Girkin, John M.

    2014-03-01

    Since their invention by Ashkin optical tweezers have demonstrated their ability and versatility as a non-invasive tool for micromanipulation. One of the most useful additions to the basic optical tweezers system is micro-Raman spectroscopy, which permits highly sensitive analysis of single cells or particles. We report on the development of a dual laser system combining two spatial light modulators to holographically manipulate multiple traps (at 1064nm) whilst undertaking Raman spectroscopy using a 532nm laser. We can thus simultaneously trap multiple particles and record their Raman spectra, without perturbing the trapping system. The dual beam system is built around micro-fluidic channels where crystallisation of calcium carbonate occurs on polymethylmethacrylate (PMMA) beads. The setup is designed to simulate at a microscopic level the reactions that occur on items in a dishwasher, where permanent filming of calcium carbonate on drinking glasses is a problem. Our system allows us to monitor crystal growth on trapped particles in which the Raman spectrum and changes in movement of the bead are recorded. Due to the expected low level of crystallisation on the bead surfaces this allows us to obtain results quickly and with high sensitivity. The long term goal is to study the development of filming on samples in-situ with the microfl.uidic system acting as a model dishwasher.

  3. [Simultaneous determination of four compounds in Sanjing Shuanghuanglian Oral Liquid by high performance liquid chromatography-diode array detection-electrochemical detection].

    Science.gov (United States)

    Liu, Lin; Suo, Zhirong; Zheng, Jianbin

    2006-05-01

    Chlorogenic acid, caffeic acid, baicalin and luteolin in Sanjing Shuanghuanglian Oral Liquid were simultaneously detected and identified using a high performance liquid chromatography coupled with diode array detection and electrochemical detection (HPLC-DAD-ECD). The separation was performed on a Zorbax SB-C18 column (150 mm x 4.6 mm i. d., 5.0 microm). The mobile phase consisted of (A) methanol and (B) methanol-water-acetic acid (50: 50: 1, v/v/v) using a linear gradient elution of 2%A-3%A at 0-3 min, 3%A-25%A at 3-15 min, 25%A-80%A at 15-20 min. The flow rate was 0.8 mL/min. The DAD detection was used at 275 nm. The ECD detection was done at 0.7 V. The column thermostat set at 30 degrees C. The limits of detection of the 4 compounds were 1 mg/L for chlorogenic acid, 0.2 mg/L for caffeic acid, 9 mg/L for baicalin, 7 mg/L for luteolin. The average recoveries were between 96.6%-99.6% with relative standard deviations (RSDs) of 2.5%-4.1%. The method is simple, rapid, reproducible and accurate. It can be used for the routine analysis of the four compounds in Shuanghuanglian Oral Liquid.

  4. Simultaneous Detection of Three Bacterial Seed-Borne Diseases in Rice Using Multiplex Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    In Jeong Kang

    2016-12-01

    Full Text Available Burkholderia glumae (bacterial grain rot, Xanthomonas oryzae pv. oryzae (bacterial leaf blight, and Acidovorax avenae subsp. avenae (bacterial brown stripe are major seedborne pathogens of rice. Based on the 16S and 23S rDNA sequences for A. avenae subsp. avenae and B. glumae, and transposase A gene sequence for X. oryzae pv. oryzae, three sets of primers had been designed to produce 402 bp for B. glumae, 490 bp for X. oryzae, and 290 bp for A. avenae subsp. avenae with the 63°C as an optimum annealing temperature. Samples collected from naturally infected fields were detected with two bacteria, B. glumae and A. avenae subsp. avenae but X. oryzae pv. oryzae was not detected. This assay can be used to identify pathogens directly from infected seeds, and will be an effective tool for the identification of the three pathogens in rice plants.

  5. Simultaneous Detection of Three Bacterial Seed-Borne Diseases in Rice Using Multiplex Polymerase Chain Reaction.

    Science.gov (United States)

    Kang, In Jeong; Kang, Mi-Hyung; Noh, Tae-Hwan; Shim, Hyeong Kwon; Shin, Dong Bum; Heu, Suggi

    2016-12-01

    Burkholderia glumae (bacterial grain rot), Xanthomonas oryzae pv. oryzae (bacterial leaf blight), and Acidovorax avenae subsp. avenae (bacterial brown stripe) are major seedborne pathogens of rice. Based on the 16S and 23S rDNA sequences for A. avenae subsp. avenae and B. glumae , and transposase A gene sequence for X. oryzae pv. oryzae , three sets of primers had been designed to produce 402 bp for B. glumae , 490 bp for X. oryzae , and 290 bp for A. avenae subsp. avenae with the 63°C as an optimum annealing temperature. Samples collected from naturally infected fields were detected with two bacteria, B. glumae and A. avenae subsp. avenae but X. oryzae pv. oryzae was not detected. This assay can be used to identify pathogens directly from infected seeds, and will be an effective tool for the identification of the three pathogens in rice plants.

  6. A Laboratory-Developed TaqMan Array Card for Simultaneous Detection of 19 Enteropathogens

    Science.gov (United States)

    Liu, Jie; Gratz, Jean; Amour, Caroline; Kibiki, Gibson; Becker, Stephen; Janaki, Lalitha; Verweij, Jaco J.; Taniuchi, Mami; Sobuz, Shihab U.; Haque, Rashidul; Haverstick, Doris M.

    2013-01-01

    The TaqMan Array Card (TAC) system is a 384-well singleplex real-time PCR format that has been used to detect multiple infection targets. Here we developed an enteric TaqMan Array Card to detect 19 enteropathogens, including viruses (adenovirus, astrovirus, norovirus GII, rotavirus, and sapovirus), bacteria (Campylobacter jejuni/C. coli, Clostridium difficile, Salmonella, Vibrio cholerae, diarrheagenic Escherichia coli strains including enteroaggregative E. coli [EAEC], enterotoxigenic E. coli [ETEC], enteropathogenic E. coli [EPEC], and Shiga-toxigenic E. coli [STEC]), Shigella/enteroinvasive E. coli (EIEC), protozoa (Cryptosporidium, Giardia lamblia, and Entamoeba histolytica), and helminths (Ascaris lumbricoides and Trichuris trichiura), as well as two extrinsic controls to monitor extraction and amplification efficiency (the bacteriophage MS2 and phocine herpesvirus). Primers and probes were newly designed or adapted from published sources and spotted onto microfluidic cards. Fecal samples were spiked with extrinsic controls, and DNA and RNA were extracted using the QiaAmp Stool DNA minikit and the QuickGene RNA Tissue kit, respectively, and then mixed with Ag-Path-ID One Step real-time reverse transcription-PCR (RT-PCR) reagents and loaded into cards. PCR efficiencies were between 90% and 105%, with linearities of 0.988 to 1. The limit of detection of the assays in the TAC was within a 10-fold difference from the cognate assays performed on plates. Precision testing demonstrated a coefficient of variation of below 5% within a run and 14% between runs. Accuracy was evaluated for 109 selected clinical specimens and revealed an average sensitivity and specificity of 85% and 77%, respectively, compared with conventional methods (including microscopy, culture, and immunoassay) and 98% and 96%, respectively, compared with our laboratory-developed PCR-Luminex assays. This TAC allows fast, accurate, and quantitative detection of a broad spectrum of enteropathogens and

  7. Simultaneous determination of organophosphorous insecticides in bean samples by gas chromatography - flame photometric detection

    Directory of Open Access Journals (Sweden)

    Keyller Bastos Borges

    2014-02-01

    Full Text Available The indiscriminate use of organophosphorous pesticides (OPPs in crops may leave residues in food and may cause poisoning in the applicators. A method was developed for the determination of five OPPs in bean samples by Gas Chromatography-Flame Photometric Detection (GC-FPD. Validation parameters comprised linearity between 0.24 and 8.56 μg g-1 (r = 0.9985 for diazinon; 0.23 and 8.14 μg g-1 (r = 0.9959 for methyl parathion; 0.28 and 10.25 μg g-1 (r = 0.9987 for methyl pirimiphos; 0.52 and 18.87 μg g-1 (r = 0.9955 for malathion; 0.86 and 13.67 μg g-1 (r = 0.9919 for ethion. The limits of quantification (equal to those of detection were the lowest rates of ranges mentioned above for each compound. The extraction method showed approximately 95% recovery, with CV% < 15%. Although twenty-eight bean samples obtained in the southern region of the state of Minas Gerais,Brazil, were analyzed, they failed to match any of the OPPs under analysis. The absence of OPPs in the samples could be due to the degradation that occurred between the use of OPPs and bean commercialization, levels below the detection /quantification limits and the non-use of OPPs in bean cultivation.

  8. Simultaneous detection of multiple mycotoxins in broiler feeds using a liquid chromatography tandem-mass spectrometry.

    Science.gov (United States)

    Kongkapan, Jutamart; Poapolathep, Saranya; Isariyodom, Supaporn; Kumagai, Susumu; Poapolathep, Amnart

    2016-02-01

    Mycotoxins are secondary fungal metabolites that are typically present in grain and feed ingredients used for animal feeds. An analytical method using LC-ESI-MS/MS was developed to quantify nine mycotoxins, consisting of aflatoxin B1 (AFB1), AFB2, AFG1, AFG2, T-2 toxin, deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEA) and ochratoxin A (OTA) in broiler feeds. In total, 100 samples of broiler feeds were collected from poultry farms in Central Thailand. The survey found that AFB1 and ZEA were the most prevalent mycotoxins in the feed samples at percentages of 93% and 63%, respectively. The limit of detections (LODs) of investigated mycotoxins was 0.20-0.78 ng/g. AFB2, DON, AFG1, NIV and T-2 toxin were also detectable at low contamination levels with percentages of 20%, 9%, 7%, 5% and 1%, respectively, whereas OTA and AFG2 were not detected in any of the feed samples. These results suggest that there is a very low level of risk of the exposure to mycotoxins in feeds obtained from broiler farms in Central Thailand.

  9. Towards achieving a reliable leakage detection and localization algorithm for application in water piping networks: an overview

    CSIR Research Space (South Africa)

    Adedeji, KB

    2017-09-01

    Full Text Available Leakage detection and localization in pipelines has become an important aspect of water management systems. Since monitoring leakage in large-scale water distribution networks (WDNs) is a challenging task, the need to develop a reliable and robust...

  10. Ultrasensitive and simultaneous detection of heavy metal ions based on three-dimensional graphene-carbon nanotubes hybrid electrode materials

    International Nuclear Information System (INIS)

    Huang, Hui; Chen, Ting; Liu, Xiuyu; Ma, Houyi

    2014-01-01

    Highlights: • Three-dimensional graphene-MWCNTs nanocomposites were prepared. • Graphene-MWCNTs based electrochemical sensor was used to detect heavy metal ions for the first time. • The proposed sensor was certified capable for real sample with satisfactory results. - Abstract: A green and facile method was developed to prepare a novel hybrid nanocomposite that consisted of one-dimensional multi-walled carbon nanotubes (MWCNTs) and two-dimensional graphene oxide (GO) sheets. The as-prepared three-dimensional GO–MWCNTs hybrid nanocomposites exhibit excellent water-solubility owing to the high hydrophilicity of GO components; meanwhile, a certain amount of MWCNTs loaded on the surface of GO sheets through π–π interaction seem to be “dissolved” in water. Moreover, the graphene(G)-MWCNTs nanocomposites with excellent conductivity were obtained conveniently by the direct electrochemical reduction of GO–MWCNTs nanocomposites. Seeing that there is a good synergistic effect between MWCNTs and graphene components in enhancing preconcentration efficiency of metal ions and accelerating electron transfer rate at G-MWCNTs/electrolyte interface, the G-MWCNTs nanocomposites possess fast, simultaneous and sensitive detection performance for trace amounts of heavy metal ions. The electrochemical results demonstrate that the G-MWCNTs nanocomposites can act as a kind of practical sensing material to simultaneously determine Pb 2+ and Cd 2+ ions in terms of anodic stripping voltammetry (ASV). The linear calibration plots for Pb 2+ and Cd 2+ ranged from 0.5 μg L −1 to 30 μg L −1 . The detection limits were determined to be 0.2 μg L −1 (S/N = 3) for Pb 2+ and 0.1 μg L −1 (S/N = 3) for Cd 2+ in the case of a deposition time of 180 s. It is worth mentioning that the G-MWCNTs modified electrodes were successfully applied to the simultaneous detection of Cd 2+ and Pb 2+ ions in real electroplating effluent samples containing lots of surface active impurities

  11. Ultrasensitive and simultaneous detection of heavy metal ions based on three-dimensional graphene-carbon nanotubes hybrid electrode materials

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Hui; Chen, Ting [Key Laboratory for Colloid and Interface Chemistry of State Education Ministry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100 (China); Liu, Xiuyu [Shandong Academy of Sciences, Jinan 250114 (China); Ma, Houyi, E-mail: hyma@sdu.edu.cn [Key Laboratory for Colloid and Interface Chemistry of State Education Ministry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100 (China)

    2014-12-10

    Highlights: • Three-dimensional graphene-MWCNTs nanocomposites were prepared. • Graphene-MWCNTs based electrochemical sensor was used to detect heavy metal ions for the first time. • The proposed sensor was certified capable for real sample with satisfactory results. - Abstract: A green and facile method was developed to prepare a novel hybrid nanocomposite that consisted of one-dimensional multi-walled carbon nanotubes (MWCNTs) and two-dimensional graphene oxide (GO) sheets. The as-prepared three-dimensional GO–MWCNTs hybrid nanocomposites exhibit excellent water-solubility owing to the high hydrophilicity of GO components; meanwhile, a certain amount of MWCNTs loaded on the surface of GO sheets through π–π interaction seem to be “dissolved” in water. Moreover, the graphene(G)-MWCNTs nanocomposites with excellent conductivity were obtained conveniently by the direct electrochemical reduction of GO–MWCNTs nanocomposites. Seeing that there is a good synergistic effect between MWCNTs and graphene components in enhancing preconcentration efficiency of metal ions and accelerating electron transfer rate at G-MWCNTs/electrolyte interface, the G-MWCNTs nanocomposites possess fast, simultaneous and sensitive detection performance for trace amounts of heavy metal ions. The electrochemical results demonstrate that the G-MWCNTs nanocomposites can act as a kind of practical sensing material to simultaneously determine Pb{sup 2+} and Cd{sup 2+} ions in terms of anodic stripping voltammetry (ASV). The linear calibration plots for Pb{sup 2+} and Cd{sup 2+} ranged from 0.5 μg L{sup −1} to 30 μg L{sup −1}. The detection limits were determined to be 0.2 μg L{sup −1} (S/N = 3) for Pb{sup 2+} and 0.1 μg L{sup −1} (S/N = 3) for Cd{sup 2+} in the case of a deposition time of 180 s. It is worth mentioning that the G-MWCNTs modified electrodes were successfully applied to the simultaneous detection of Cd{sup 2+} and Pb{sup 2+} ions in real electroplating

  12. A Multiplex PCR for the Simultaneous Detection and Genotyping of the Echinococcus granulosus Complex

    Science.gov (United States)

    Boubaker, Ghalia; Macchiaroli, Natalia; Prada, Laura; Cucher, Marcela A.; Rosenzvit, Mara C.; Ziadinov, Iskender; Deplazes, Peter; Saarma, Urmas; Babba, Hamouda; Gottstein, Bruno; Spiliotis, Markus

    2013-01-01

    Echinococcus granulosus is characterized by high intra-specific variability (genotypes G1–G10) and according to the new molecular phylogeny of the genus Echinococcus, the E. granulosus complex has been divided into E. granulosus sensu stricto (G1–G3), E. equinus (G4), E. ortleppi (G5), and E. canadensis (G6–G10). The molecular characterization of E. granulosus isolates is fundamental to understand the spatio-temporal epidemiology of this complex in many endemic areas with the simultaneous occurrence of different Echinococcus species and genotypes. To simplify the genotyping of the E. granulosus complex we developed a single-tube multiplex PCR (mPCR) allowing three levels of discrimination: (i) Echinococcus genus, (ii) E. granulosus complex in common, and (iii) the specific genotype within the E. granulosus complex. The methodology was established with known DNA samples of the different strains/genotypes, confirmed on 42 already genotyped samples (Spain: 22 and Bulgaria: 20) and then successfully applied on 153 unknown samples (Tunisia: 114, Algeria: 26 and Argentina: 13). The sensitivity threshold of the mPCR was found to be 5 ng Echinoccoccus DNA in a mixture of up to 1 µg of foreign DNA and the specificity was 100% when template DNA from closely related members of the genus Taenia was used. Additionally to DNA samples, the mPCR can be carried out directly on boiled hydatid fluid or on alkaline-lysed frozen or fixed protoscoleces, thus avoiding classical DNA extractions. However, when using Echinococcus eggs obtained from fecal samples of infected dogs, the sensitivity of the mPCR was low (Echinococcus genus. PMID:23350011

  13. A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex.

    Science.gov (United States)

    Boubaker, Ghalia; Macchiaroli, Natalia; Prada, Laura; Cucher, Marcela A; Rosenzvit, Mara C; Ziadinov, Iskender; Deplazes, Peter; Saarma, Urmas; Babba, Hamouda; Gottstein, Bruno; Spiliotis, Markus

    2013-01-01

    Echinococcus granulosus is characterized by high intra-specific variability (genotypes G1-G10) and according to the new molecular phylogeny of the genus Echinococcus, the E. granulosus complex has been divided into E. granulosus sensu stricto (G1-G3), E. equinus (G4), E. ortleppi (G5), and E. canadensis (G6-G10). The molecular characterization of E. granulosus isolates is fundamental to understand the spatio-temporal epidemiology of this complex in many endemic areas with the simultaneous occurrence of different Echinococcus species and genotypes. To simplify the genotyping of the E. granulosus complex we developed a single-tube multiplex PCR (mPCR) allowing three levels of discrimination: (i) Echinococcus genus, (ii) E. granulosus complex in common, and (iii) the specific genotype within the E. granulosus complex. The methodology was established with known DNA samples of the different strains/genotypes, confirmed on 42 already genotyped samples (Spain: 22 and Bulgaria: 20) and then successfully applied on 153 unknown samples (Tunisia: 114, Algeria: 26 and Argentina: 13). The sensitivity threshold of the mPCR was found to be 5 ng Echinoccoccus DNA in a mixture of up to 1 µg of foreign DNA and the specificity was 100% when template DNA from closely related members of the genus Taenia was used. Additionally to DNA samples, the mPCR can be carried out directly on boiled hydatid fluid or on alkaline-lysed frozen or fixed protoscoleces, thus avoiding classical DNA extractions. However, when using Echinococcus eggs obtained from fecal samples of infected dogs, the sensitivity of the mPCR was low (Echinococcus genus.

  14. A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex.

    Directory of Open Access Journals (Sweden)

    Ghalia Boubaker

    Full Text Available Echinococcus granulosus is characterized by high intra-specific variability (genotypes G1-G10 and according to the new molecular phylogeny of the genus Echinococcus, the E. granulosus complex has been divided into E. granulosus sensu stricto (G1-G3, E. equinus (G4, E. ortleppi (G5, and E. canadensis (G6-G10. The molecular characterization of E. granulosus isolates is fundamental to understand the spatio-temporal epidemiology of this complex in many endemic areas with the simultaneous occurrence of different Echinococcus species and genotypes. To simplify the genotyping of the E. granulosus complex we developed a single-tube multiplex PCR (mPCR allowing three levels of discrimination: (i Echinococcus genus, (ii E. granulosus complex in common, and (iii the specific genotype within the E. granulosus complex. The methodology was established with known DNA samples of the different strains/genotypes, confirmed on 42 already genotyped samples (Spain: 22 and Bulgaria: 20 and then successfully applied on 153 unknown samples (Tunisia: 114, Algeria: 26 and Argentina: 13. The sensitivity threshold of the mPCR was found to be 5 ng Echinoccoccus DNA in a mixture of up to 1 µg of foreign DNA and the specificity was 100% when template DNA from closely related members of the genus Taenia was used. Additionally to DNA samples, the mPCR can be carried out directly on boiled hydatid fluid or on alkaline-lysed frozen or fixed protoscoleces, thus avoiding classical DNA extractions. However, when using Echinococcus eggs obtained from fecal samples of infected dogs, the sensitivity of the mPCR was low (<40%. Thus, except for copro analysis, the mPCR described here has a high potential for a worldwide application in large-scale molecular epidemiological studies on the Echinococcus genus.

  15. Simultaneous Detection of Four Foodborne Viruses in Food Samples Using a One-Step Multiplex Reverse Transcription PCR.

    Science.gov (United States)

    Lee, Shin-Young; Kim, Mi-Ju; Kim, Hyun-Joong; Jeong, KwangCheol Casey; Kim, Hae-Yeong

    2018-02-28

    A one-step multiplex reverse transcription PCR (RT-PCR) method comprising six primer sets (for the detection of norovirus GI and GII, hepatitis A virus, rotavirus, and astrovirus) was developed to simultaneously detect four kinds of pathogenic viruses. The size of the PCR products for norovirus GI and GII, hepatitis A virus (VP3/VP1 and P2A regions), rotavirus, and astrovirus were 330, 164, 244, 198, 629, and 449 bp, respectively. The RT-PCR with the six primer sets showed specificity for the pathogenic viruses. The detection limit of the developed multiplex RT-PCR, as evaluated using serially diluted viral RNAs, was comparable to that of one-step single RT-PCR. Moreover, this multiplex RT-PCR was evaluated using food samples such as water, oysters, lettuce, and vegetable product. These food samples were artificially spiked with the four kinds of viruses in diverse combinations, and the spiked viruses in all food samples were detected successfully.

  16. Simultaneous detection of nine cyanotoxins in drinking water using dual solid-phase extraction and liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Yen, Hung-Kai; Lin, Tsair-Fuh; Liao, Pao-Chi

    2011-08-01

    A solid-phase extraction (SPE)-liquid chromatography (LC)-mass spectrometry (MS) method was developed to concentrate and detect nine cyanotoxins simultaneously, including six microcystins (MCs) congeners, nodularin (NOD), anatoxin-a (ATX) and cylindrospermopsin (CYN), in pure and natural waters. A dual cartridge SPE assembly was tested for the operating parameters of cyanotoxin extraction. A surrogate standard (SS), 1,9-diaminononane, was spiked in all the samples before the SPE extraction, and an internal standard (IS), 2,3,5-trimethylphenyl methyl carbamate, was spiked before LC/MS analysis. The method detection limit (MDL) was 2-100 ng/L for nine cyanotoxins in pure water and was increased by a factor of three to ten in a more complicated water matrix. The recoveries based on SS were between 83 and 104%, while those based on IS were 80-120%. The developed method was successfully employed in analyzing 33 water samples collected from eutrophic lakes, water treatment plants and distribution taps. MCs, NOD, and CYN were detected in the reservoir water, with concentrations as high as 36 μg/L. In addition, for the first time in Taiwan's tap water, CYN was detected at concentrations as high as 8.6 μg/L. Quality control data for the field samples shows that the analytical scheme developed is appropriate for monitoring cyanotoxins. Copyright © 2011 Elsevier Ltd. All rights reserved.

  17. Simultaneous Detection of α-Fetoprotein and Carcinoembryonic Antigen Based on Si Nanowire Field-Effect Transistors

    Directory of Open Access Journals (Sweden)

    Kuiyu Zhu

    2015-08-01

    Full Text Available Primary hepatic carcinoma (PHC is one of the most common malignancies worldwide, resulting in death within six to 20 months. The survival rate can be improved by effective treatments when diagnosed at an early stage. The α-fetoprotein (AFP and carcinoembryonic antigen (CEA have been identified as markers that are expressed at higher levels in PHC patients. In this study, we employed silicon nanowire field-effect transistors (SiNW-FETs with polydimethylsiloxane (PDMS microfluidic channels to simultaneously detect AFP and CEA in desalted human serum. Dual-channel PDMS was first utilized for the selective modification of AFP and CEA antibodies on SiNWs, while single-channel PDMS offers faster and more sensitive detection of AFP and CEA in serum. During the SiNW modification process, 0.1% BSA was utilized to minimize nonspecific protein binding from serum. The linear dynamic ranges for the AFP and CEA detection were measured to be 500 fg/mL to 50 ng/mL and 50 fg/mL to 10 ng/mL, respectively. Our work demonstrates the promising potential of fabricated SiNW-FETs as a direct detection kit for multiple tumor markers in serum; therefore, it provides a chance for early stage diagnose and, hence, more effective treatments for PHC patients.

  18. Simultaneous detection of landmarks and key-frame in cardiac perfusion MRI using a joint spatial-temporal context model

    Science.gov (United States)

    Lu, Xiaoguang; Xue, Hui; Jolly, Marie-Pierre; Guetter, Christoph; Kellman, Peter; Hsu, Li-Yueh; Arai, Andrew; Zuehlsdorff, Sven; Littmann, Arne; Georgescu, Bogdan; Guehring, Jens

    2011-03-01

    Cardiac perfusion magnetic resonance imaging (MRI) has proven clinical significance in diagnosis of heart diseases. However, analysis of perfusion data is time-consuming, where automatic detection of anatomic landmarks and key-frames from perfusion MR sequences is helpful for anchoring structures and functional analysis of the heart, leading toward fully automated perfusion analysis. Learning-based object detection methods have demonstrated their capabilities to handle large variations of the object by exploring a local region, i.e., context. Conventional 2D approaches take into account spatial context only. Temporal signals in perfusion data present a strong cue for anchoring. We propose a joint context model to encode both spatial and temporal evidence. In addition, our spatial context is constructed not only based on the landmark of interest, but also the landmarks that are correlated in the neighboring anatomies. A discriminative model is learned through a probabilistic boosting tree. A marginal space learning strategy is applied to efficiently learn and search in a high dimensional parameter space. A fully automatic system is developed to simultaneously detect anatomic landmarks and key frames in both RV and LV from perfusion sequences. The proposed approach was evaluated on a database of 373 cardiac perfusion MRI sequences from 77 patients. Experimental results of a 4-fold cross validation show superior landmark detection accuracies of the proposed joint spatial-temporal approach to the 2D approach that is based on spatial context only. The key-frame identification results are promising.

  19. Simultaneous detection of mutations and copy number variation of NPM1 in the acute myeloid leukemia using multiplex ligation-dependent probe amplification

    International Nuclear Information System (INIS)

    Marcinkowska-Swojak, Malgorzata; Handschuh, Luiza; Wojciechowski, Pawel; Goralski, Michal; Tomaszewski, Kamil; Kazmierczak, Maciej; Lewandowski, Krzysztof; Komarnicki, Mieczyslaw

    2016-01-01

    Highlights: • The NPM1 mutations were detected exclusively in AML accounting for 25% of cases. • The NPM1 gene did not reveal any copy number alterations. • The NPM1mut+ assay is a reliable test for the analysis of mutations and CNA in NPM1. - Abstract: The NPM1 gene encodes nucleophosmin, a protein involved in multiple cell functions and carcinogenesis. Mutation of the NPM1 gene, causing delocalization of the protein, is the most frequent genetic lesion in acute myeloid leukemia (AML); it is considered a founder event in AML pathogenesis and serves as a favorable prognostic marker. Moreover, in solid tumors and some leukemia cell lines, overexpression of the NPM1 gene is commonly observed. Therefore, the purpose of this study was to develop a new method for the detection of NPM1 mutations and the simultaneous analysis of copy number alterations (CNAs), which may underlie NPM1 gene expression deregulation. To address both of the issues, we applied a strategy based on multiplex ligation-dependent probe amplification (MLPA). A designed NPM1mut+ assay enables the detection of three of the most frequent NPM1 mutations: A, B and D. The accuracy of the assay was tested using a group of 83 samples from Polish patients with AML and other blood-proliferative disorders. To verify the results, we employed traditional Sanger sequencing and next-generation transcriptome sequencing. With the use of the NPM1mut+ assay, we detected mutations A, D and B in 14, 1 and 0 of the analyzed samples, respectively. All of these mutations were confirmed by complementary sequencing approaches, proving the 100% specificity and sensitivity of the proposed test. The performed sequencing analysis allowed the identification of two additional rare mutations (I and ZE). All of the mutations were identified exclusively in AML cases, accounting for 25% of those cases. We did not observe any CNAs (amplifications) of the NPM1 gene in the studied samples, either with or without the mutation. The

  20. Simultaneous detection of mutations and copy number variation of NPM1 in the acute myeloid leukemia using multiplex ligation-dependent probe amplification

    Energy Technology Data Exchange (ETDEWEB)

    Marcinkowska-Swojak, Malgorzata, E-mail: m-marcinkowska@o2.pl [European Center of Bioinformatics and Genomics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Z. Noskowskiego 12/14, 61-704 Poznan (Poland); Handschuh, Luiza, E-mail: luizahan@ibch.poznan.pl [European Center of Bioinformatics and Genomics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Z. Noskowskiego 12/14, 61-704 Poznan (Poland); Department of Hematology and Bone Marrow Transplantation, Poznan University of Medical Sciences, Szamarzewskiego 82/84, 60-569 Poznan (Poland); Wojciechowski, Pawel, E-mail: Pawel.Wojciechowski@cs.put.poznan.pl [European Center of Bioinformatics and Genomics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Z. Noskowskiego 12/14, 61-704 Poznan (Poland); Institute of Computing Science, Poznan University of Technology, Piotrowo 2, 60-965 Poznan (Poland); Goralski, Michal, E-mail: mgoralsk@ibch.poznan.pl [European Center of Bioinformatics and Genomics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Z. Noskowskiego 12/14, 61-704 Poznan (Poland); Tomaszewski, Kamil, E-mail: kamil.tomaszewsky@gmail.com [European Center of Bioinformatics and Genomics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Z. Noskowskiego 12/14, 61-704 Poznan (Poland); Kazmierczak, Maciej, E-mail: maciej.kazmierczak@onet.eu [Department of Hematology and Bone Marrow Transplantation, Poznan University of Medical Sciences, Szamarzewskiego 82/84, 60-569 Poznan (Poland); Lewandowski, Krzysztof, E-mail: krzysztof.lewandowski@skpp.edu.pl [Department of Hematology and Bone Marrow Transplantation, Poznan University of Medical Sciences, Szamarzewskiego 82/84, 60-569 Poznan (Poland); Komarnicki, Mieczyslaw, E-mail: mak7@pro.onet.pl [Department of Hematology and Bone Marrow Transplantation, Poznan University of Medical Sciences, Szamarzewskiego 82/84, 60-569 Poznan (Poland); and others

    2016-04-15

    Highlights: • The NPM1 mutations were detected exclusively in AML accounting for 25% of cases. • The NPM1 gene did not reveal any copy number alterations. • The NPM1mut+ assay is a reliable test for the analysis of mutations and CNA in NPM1. - Abstract: The NPM1 gene encodes nucleophosmin, a protein involved in multiple cell functions and carcinogenesis. Mutation of the NPM1 gene, causing delocalization of the protein, is the most frequent genetic lesion in acute myeloid leukemia (AML); it is considered a founder event in AML pathogenesis and serves as a favorable prognostic marker. Moreover, in solid tumors and some leukemia cell lines, overexpression of the NPM1 gene is commonly observed. Therefore, the purpose of this study was to develop a new method for the detection of NPM1 mutations and the simultaneous analysis of copy number alterations (CNAs), which may underlie NPM1 gene expression deregulation. To address both of the issues, we applied a strategy based on multiplex ligation-dependent probe amplification (MLPA). A designed NPM1mut+ assay enables the detection of three of the most frequent NPM1 mutations: A, B and D. The accuracy of the assay was tested using a group of 83 samples from Polish patients with AML and other blood-proliferative disorders. To verify the results, we employed traditional Sanger sequencing and next-generation transcriptome sequencing. With the use of the NPM1mut+ assay, we detected mutations A, D and B in 14, 1 and 0 of the analyzed samples, respectively. All of these mutations were confirmed by complementary sequencing approaches, proving the 100% specificity and sensitivity of the proposed test. The performed sequencing analysis allowed the identification of two additional rare mutations (I and ZE). All of the mutations were identified exclusively in AML cases, accounting for 25% of those cases. We did not observe any CNAs (amplifications) of the NPM1 gene in the studied samples, either with or without the mutation. The

  1. Simultaneous detection and removal of radioisotopes with modified alginate beads containing an azo-based probe using RGB coordinates

    International Nuclear Information System (INIS)

    Jo, Ara; Jang, Geunseok; Namgung, Ho; Kim, Choongho; Kim, Daigeun; Kim, Yujun; Kim, Jongho; Lee, Taek Seung

    2015-01-01

    Highlights: • Modified alginate with azo-based probe (ABO) was synthesized by a reaction between sodium alginate and azo-based probe (BO2). • BO2 was found to be a good probe molecule for radioisotopes using colorimetric analysis. • Detection of Co 2+ and Sr 2+ was mainly carried out via interaction between BO2 and metal ions. • Simultaneous removal of radioisotopes was assessed by the ion-exchange of carboxylate groups in sodium alginate. • The alginate beads with dual functions of detection and removal of metal ions are successfully accomplished. - Abstract: We prepared alginate beads that were modified with an azo-based probe molecule to monitor simultaneously the removal (by alginate) and probing (by the azo-probe molecule) of radioisotopes such as cobalt, strontium, and cesium ions. As an azo-probe molecule, Basic Orange 2 (BO2) was immobilized to the alginate bead. The BO2 in aqueous solution exhibited a slight red shift in absorption with a change in color from orange to dark orange upon addition of cobalt and strontium ions. In contrast, the color of BO2 did not change upon exposure to cesium ions. Thus, the covalently embedded BO2 in alginate beads could adsorb cobalt and strontium ions resulting in recognizable color change of the beads, which was induced by the formation of a complex between BO2 and metal ions. The color changes of the beads in the presence of metal ions were determined quantitatively using RGB color coordinate values. In addition to effectively removing metal ions, the colorimetric coordinate method provides a convenient and simple sensing technique for naked-eye metal ion detection.

  2. A convenient ultrasound-assisted saponification for the simultaneous determination of vitamin E isomers in vegetable oil by HPLC with fluorescence detection.

    Science.gov (United States)

    Yang, Yi; Lu, Dan; Yin, Shuo; Yang, Danni; Chen, Yaling; Li, Yongxin; Sun, Chengjun

    2018-04-01

    An efficient ultrasound-assisted saponification was developed for simultaneous determination of vitamin E isomers in vegetable oil by high-performance liquid chromatography with fluorescence detection. The samples were saponified ultrasonically with potassium hydroxide solution for only 7 min, then the analytes were extracted with ether. Vitamin E isomers were separated on a C 18 column at 25°C with a mobile phase of methanol/acetonitrile (81:19, v/v) at a flow rate of 0.8 mL/min. Fluorescence detection was operated at 290 nm of excitation wavelength and 327 nm of emission wavelength. Under the optimized conditions, good linearities over the range of 0.001-8.00 μg/mL (r > 0.999) were obtained. Mean recoveries of the method were 88.0-106%, with intra- and interday RSDs less than 11.8 and 12.8%, respectively. The detection limits and quantification limits of the method were 0.30-1.8 and 1.0-6.1 μg/kg, respectively. The recoveries of this method were much higher than that of the quick, easy, cheap, effective, rugged, and safe method and direct dilution method, but were similar to those of hot saponification. This proposed method provides reliable, simple, and rapid quantification of vitamin E isomers in vegetable oils. Five kinds of vegetable oils were analyzed, the quantification results were within the ranges reported by other authors. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Multiplex Real-Time qPCR Assay for Simultaneous and Sensitive Detection of Phytoplasmas in Sesame Plants and Insect Vectors.

    Science.gov (United States)

    Ikten, Cengiz; Ustun, Rustem; Catal, Mursel; Yol, Engin; Uzun, Bulent

    2016-01-01

    Phyllody, a destructive and economically important disease worldwide caused by phytoplasma infections, is characterized by the abnormal development of floral structures into stunted leafy parts and contributes to serious losses in crop plants, including sesame (Sesamum indicum L.). Accurate identification, differentiation, and quantification of phyllody-causing phytoplasmas are essential for effective management of this plant disease and for selection of resistant sesame varieties. In this study, a diagnostic multiplex qPCR assay was developed using TaqMan® chemistry based on detection of the 16S ribosomal RNA gene of phytoplasmas and the 18S ribosomal gene of sesame. Phytoplasma and sesame specific primers and probes labeled with different fluorescent dyes were used for simultaneous amplification of 16SrII and 16SrIX phytoplasmas in a single tube. The multiplex real-time qPCR assay allowed accurate detection, differentiation, and quantification of 16SrII and 16SrIX groups in 109 sesame plant and 92 insect vector samples tested. The assay was found to have a detection sensitivity of 1.8 x 102 and 1.6 x 102 DNA copies for absolute quantification of 16SrII and 16SrIX group phytoplasmas, respectively. Relative quantification was effective and reliable for determination of phyllody phytoplasma DNA amounts normalized to sesame DNA in infected plant tissues. The development of this qPCR assay provides a method for the rapid measurement of infection loads to identify resistance levels of sesame genotypes against phyllody phytoplasma disease.

  4. Multiplex Real-Time qPCR Assay for Simultaneous and Sensitive Detection of Phytoplasmas in Sesame Plants and Insect Vectors.

    Directory of Open Access Journals (Sweden)

    Cengiz Ikten

    Full Text Available Phyllody, a destructive and economically important disease worldwide caused by phytoplasma infections, is characterized by the abnormal development of floral structures into stunted leafy parts and contributes to serious losses in crop plants, including sesame (Sesamum indicum L.. Accurate identification, differentiation, and quantification of phyllody-causing phytoplasmas are essential for effective management of this plant disease and for selection of resistant sesame varieties. In this study, a diagnostic multiplex qPCR assay was developed using TaqMan® chemistry based on detection of the 16S ribosomal RNA gene of phytoplasmas and the 18S ribosomal gene of sesame. Phytoplasma and sesame specific primers and probes labeled with different fluorescent dyes were used for simultaneous amplification of 16SrII and 16SrIX phytoplasmas in a single tube. The multiplex real-time qPCR assay allowed accurate detection, differentiation, and quantification of 16SrII and 16SrIX groups in 109 sesame plant and 92 insect vector samples tested. The assay was found to have a detection sensitivity of 1.8 x 102 and 1.6 x 102 DNA copies for absolute quantification of 16SrII and 16SrIX group phytoplasmas, respectively. Relative quantification was effective and reliable for determination of phyllody phytoplasma DNA amounts normalized to sesame DNA in infected plant tissues. The development of this qPCR assay provides a method for the rapid measurement of infection loads to identify resistance levels of sesame genotypes against phyllody phytoplasma disease.

  5. Reliable Detection and Smart Deletion of Malassez Counting Chamber Grid in Microscopic White Light Images for Microbiological Applications.

    Science.gov (United States)

    Denimal, Emmanuel; Marin, Ambroise; Guyot, Stéphane; Journaux, Ludovic; Molin, Paul

    2015-08-01

    In biology, hemocytometers such as Malassez slides are widely used and are effective tools for counting cells manually. In a previous work, a robust algorithm was developed for grid extraction in Malassez slide images. This algorithm was evaluated on a set of 135 images and grids were accurately detected in most cases, but there remained failures for the most difficult images. In this work, we present an optimization of this algorithm that allows for 100% grid detection and a 25% improvement in grid positioning accuracy. These improvements make the algorithm fully reliable for grid detection. This optimization also allows complete erasing of the grid without altering the cells, which eases their segmentation.

  6. A novel approach to detect KRAS/BRAF mutation for colon cancer: Highly sensitive simultaneous detection of mutations and simple pre-treatment without DNA extraction.

    Science.gov (United States)

    Suzuki, Shun-Ichi; Matsusaka, Satoshi; Hirai, Mitsuharu; Shibata, Harumi; Takagi, Koichi; Mizunuma, Nobuyuki; Hatake, Kiyohiko

    2015-07-01

    It has been reported that colon cancer patients with KRAS and BRAF mutations that lie downstream of epidermal growth factor receptor (EGFR) acquire resistance against therapy with anti‑EGFR antibodies, cetuximab and panitumumab. On the other hand, some reports say KRAS codon 13 mutation (p.G13D) has lower resistance against anti-EGFR antibodies, thus there is a substantial need for detection of specific KRAS mutations. We have established a state-of-the-art measurement system using QProbe (QP) method that allows simultaneous measurement of KRAS codon 12/13, p.G13D and BRAF mutation, and compared this method against Direct Sequencing (DS) using 182 specimens from colon cancer patients. In addition, 32 biopsy specimens were processed with a novel pre-treatment method without DNA purification in order to detect KRAS/BRAF. As a result of KRAS mutation measurement, concordance rate between the QP method and DS method was 81.4% (144/177) except for the 5 specimens that were undeterminable. Among them, 29 specimens became positive with QP method and negative with DS method. BRAF was measured with QP method only, and the mutation detection rate was 3.9% (6/153). KRAS measurement using a simple new pre-treatment method without DNA extraction resulted in 31 good results out of 32, all of them matching with the DS method. We have established a simple but highly sensitive simultaneous detection system for KRAS/BRAF. Moreover, introduction of the novel pre-treatment technology eliminated the inconvenient DNA extraction process. From this research achievement, we not only anticipate quick and accurate results returned in the clinical field but also contribution in improving the test quality and work efficiency.

  7. Phytohormones as Important Biologically Active Molecules – Their Simple Simultaneous Detection

    Directory of Open Access Journals (Sweden)

    Ladislav Havel

    2009-05-01

    Full Text Available Phytohormones, their functions, synthesis and effects, are of great interest. To study them in plant tissues accurate and sensitive methods are required. In the present study we aimed at optimizing experimental conditions to separate and determine not only plant hormones but also their metabolites, by liquid chromatography coupled with a UV-VIS detector. The mixture we analyzed was composed of benzyladenine, kinetin, trans-zeatin, cis-zeatin, dihydrozeatin, meta-topolin, ortho-topolin, α-naphthalene acetic acid, indole-3-acetic acid, trans-zeatin-7-glucoside, trans-zeatin-O-glucoside, trans-zeatin-9-riboside, meta-topolin-9-riboside and ortho-topolin-9-riboside. We measured the calibration dependences and estimated limits of detection and quantification under the optimal chromatographic conditions (column: Polaris C18; mobile phase: gradient starting at 2:98 (methanol:0.001% TFA and was increasing to 55:45 during twenty minutes, and then decreasing for 10 min to 35:65, flow rate: 200 µL·min-1, temperature: 50 °C, wavelength: 210 nm. The detection limits for the target molecules were estimated as tens of ng per mL. We also studied the effect of flax extracts on the phytohormones’ signals. Recovery of aliphatic and aromatic cytokinins, metabolites of cytokinins and auxinswere within the range from 87 to 105 %. The experimental conditions were tested on a mass selective detector. In addition we analysed a commercial product used for stimulation of roots formation in cuttings of poorly rooting plants. The determined content of α-naphthalene acetic acid was in good agreement with that declared by the manufacturer.

  8. Simultaneous Detection of Flavonoids, Phenolic Acids and Alkaloids in Abri Herba and Abri Mollis Herba using Liquid Chromatography Tandem Mass Spectrometry.

    Science.gov (United States)

    Yan, Wenying; Han, Qingjie; Guo, Panpan; Wang, Chunying; Zhang, Zijian

    2016-01-01

    Abri Herba has remarkable properties, such as cleanup heat detoxification, dampness and activating blood circulation to dissipate blood stasis; as a result, it has been applied to treat acute or chronic hepatitis and mastitis. Abri mollis Herba is often used as Abri Herba. Hierarchical cluster analysis (HCA) was applied to compare the similarities and differences of the chemical compositions in the two types of medicinal materials. To establish a high-performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS) method for the simultaneous analysis of 15 flavonoids, two phenolic acids and three alkaloids in Abri Herba and Abri mollis Herba. The chromatographic separation was performed on a C18 column with a mobile phase of methanol (A), acetonitrile (B) and 0.5‰ acetic acid in water (C) using gradient elution. The detection of the target compounds was performed in multiple-reaction monitoring (MRM) mode using a hybrid quadrupole linear ion trap mass spectrometer equipped with positive/negative ion-switching electrospray ionisation (ESI) source. The developed method is reliable, sensitive and specific. In addition, the method has been successfully applied to differentiate 15 batches of Abri Herba and 27 batches of Abri mollis Herba stems. Furthermore, a comparison of the contents among stems, roots and leaves from the same strain in seven batches of Abri mollis Herba and four batches of Abri Herba has also been performed. HPLC-MS/MS method is sensitive and selective and can be suitable for the reliable quality control of Abri mollis Herba and Abri Herba. Copyright © 2015 John Wiley & Sons, Ltd.

  9. The reliability, accuracy and minimal detectable difference of a multi-segment kinematic model of the foot-shoe complex.

    Science.gov (United States)

    Bishop, Chris; Paul, Gunther; Thewlis, Dominic

    2013-04-01

    Kinematic models are commonly used to quantify foot and ankle kinematics, yet no marker sets or models have been proven reliable or accurate when wearing shoes. Further, the minimal detectable difference of a developed model is often not reported. We present a kinematic model that is reliable, accurate and sensitive to describe the kinematics of the foot-shoe complex and lower leg during walking gait. In order to achieve this, a new marker set was established, consisting of 25 markers applied on the shoe and skin surface, which informed a four segment kinematic model of the foot-shoe complex and lower leg. Three independent experiments were conducted to determine the reliability, accuracy and minimal detectable difference of the marker set and model. Inter-rater reliability of marker placement on the shoe was proven to be good to excellent (ICC=0.75-0.98) indicating that markers could be applied reliably between raters. Intra-rater reliability was better for the experienced rater (ICC=0.68-0.99) than the inexperienced rater (ICC=0.38-0.97). The accuracy of marker placement along each axis was <6.7 mm for all markers studied. Minimal detectable difference (MDD90) thresholds were defined for each joint; tibiocalcaneal joint--MDD90=2.17-9.36°, tarsometatarsal joint--MDD90=1.03-9.29° and the metatarsophalangeal joint--MDD90=1.75-9.12°. These thresholds proposed are specific for the description of shod motion, and can be used in future research designed at comparing between different footwear. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Reliability, validity and minimal detectable change of the Mini-BESTest in Greek participants with chronic stroke.

    Science.gov (United States)

    Lampropoulou, Sofia I; Billis, Evdokia; Gedikoglou, Ingrid A; Michailidou, Christina; Nowicky, Alexander V; Skrinou, Dimitra; Michailidi, Fotini; Chandrinou, Danae; Meligkoni, Margarita

    2018-02-23

    This study aimed to investigate the psychometric characteristics of reliability, validity and ability to detect change of a newly developed balance assessment tool, the Mini-BESTest, in Greek patients with stroke. A prospective, observational design study with test-retest measures was conducted. A convenience sample of 21 Greek patients with chronic stroke (14 male, 7 female; age of 63 ± 16 years) was recruited. Two independent examiners administered the scale, for the inter-rater reliability, twice within 10 days for the test-retest reliability. Bland Altman Analysis for repeated measures assessed the absolute reliability and the Standard Error of Measurement (SEM) and the Minimum Detectable Change at 95% confidence interval (MDC 95% ) were established. The Greek Mini-BESTest (Mini-BESTest GR ) was correlated with the Greek Berg Balance Scale (BBS GR ) for assessing the concurrent validity and with the Timed Up and Go (TUG), the Functional Reach Test (FRT) and the Greek Falls Efficacy Scale-International (FES-I GR ) for the convergent validity. The Mini-BESTestGR demonstrated excellent inter-rater reliability (ICC (95%CI) = 0.997 (0.995-0.999, SEM = 0.46) with the scores of two raters within the limits of agreement (mean dif  = -0.143 ± 0.727, p > 0.05) and test-retest reliability (ICC (95%CI) = 0.966 (0.926-0.988), SEM = 1.53). Additionally, the Mini-BESTest GR yielded very strong to moderate correlations with BBS GR (r = 0.924, p reliability and the equally good validity of the Mini-BESTest GR , strongly support its utility in Greek people with chronic stroke. Its ability to identify clinically meaningful changes and falls risk need further investigation.

  11. Simultaneous fingerprint and high-wavenumber confocal Raman spectroscopy enhances early detection of cervical precancer in vivo.

    Science.gov (United States)

    Duraipandian, Shiyamala; Zheng, Wei; Ng, Joseph; Low, Jeffrey J H; Ilancheran, A; Huang, Zhiwei

    2012-07-17

    Raman spectroscopy is a vibrational spectroscopic technique capable of nondestructively probing endogenous biomolecules and their changes associated with dysplastic transformation in the tissue. The main objectives of this study are (i) to develop a simultaneous fingerprint (FP) and high-wavenumber (HW) confocal Raman spectroscopy and (ii) to investigate its diagnostic utility for improving in vivo diagnosis of cervical precancer (dysplasia). We have successfully developed an integrated FP/HW confocal Raman diagnostic system with a ball-lens Raman probe for simultaneous acquistion of FP/HW Raman signals of the cervix in vivo within 1 s. A total of 476 in vivo FP/HW Raman spectra (356 normal and 120 precancer) are acquired from 44 patients at clinical colposcopy. The distinctive Raman spectral differences between normal and dysplastic cervical tissue are observed at ~854, 937, 1001, 1095, 1253, 1313, 1445, 1654, 2946, and 3400 cm(-1) mainly related to proteins, lipids, glycogen, nucleic acids and water content in tissue. Multivariate diagnostic algorithms developed based on partial least-squares-discriminant analysis (PLS-DA) together with the leave-one-patient-out, cross-validation yield the diagnostic sensitivities of 84.2%, 76.7%, and 85.0%, respectively; specificities of 78.9%, 73.3%, and 81.7%, respectively; and overall diagnostic accuracies of 80.3%, 74.2%, and 82.6%, respectively, using FP, HW, and integrated FP/HW Raman spectroscopic techniques for in vivo diagnosis of cervical precancer. Receiver operating characteristic (ROC) analysis further confirms the best performance of the integrated FP/HW confocal Raman technique, compared to FP or HW Raman spectroscopy alone. This work demonstrates, for the first time, that the simultaneous FP/HW confocal Raman spectroscopy has the potential to be a clinically powerful tool for improving early diagnosis and detection of cervical precancer in vivo during clinical colposcopic examination.

  12. Self-Tuning Method for Increased Obstacle Detection Reliability Based on Internet of Things LiDAR Sensor Models.

    Science.gov (United States)

    Castaño, Fernando; Beruvides, Gerardo; Villalonga, Alberto; Haber, Rodolfo E

    2018-05-10

    On-chip LiDAR sensors for vehicle collision avoidance are a rapidly expanding area of research and development. The assessment of reliable obstacle detection using data collected by LiDAR sensors has become a key issue that the scientific community is actively exploring. The design of a self-tuning methodology and its implementation are presented in this paper, to maximize the reliability of LiDAR sensors network for obstacle detection in the 'Internet of Things' (IoT) mobility scenarios. The Webots Automobile 3D simulation tool for emulating sensor interaction in complex driving environments is selected in order to achieve that objective. Furthermore, a model-based framework is defined that employs a point-cloud clustering technique, and an error-based prediction model library that is composed of a multilayer perceptron neural network, and k-nearest neighbors and linear regression models. Finally, a reinforcement learning technique, specifically a Q-learning method, is implemented to determine the number of LiDAR sensors that are required to increase sensor reliability for obstacle localization tasks. In addition, a IoT driving assistance user scenario, connecting a five LiDAR sensor network is designed and implemented to validate the accuracy of the computational intelligence-based framework. The results demonstrated that the self-tuning method is an appropriate strategy to increase the reliability of the sensor network while minimizing detection thresholds.

  13. Development of a versatile tool for the simultaneous differential detection of Pseudomonas savastanoi pathovars by End Point and Real-Time PCR.

    Science.gov (United States)

    Tegli, Stefania; Cerboneschi, Matteo; Libelli, Ilaria Marsili; Santilli, Elena

    2010-05-28

    Pseudomonas savastanoi pv. savastanoi is the causal agent of olive knot disease. The strains isolated from oleander and ash belong to the pathovars nerii and fraxini, respectively. When artificially inoculated, pv. savastanoi causes disease also on ash, and pv. nerii attacks also olive and ash. Surprisingly nothing is known yet about their distribution in nature on these hosts and if spontaneous cross-infections occur. On the other hand sanitary certification programs for olive plants, also including P. savastanoi, were launched in many countries. The aim of this work was to develop several PCR-based tools for the rapid, simultaneous, differential and quantitative detection of these P. savastanoi pathovars, in multiplex and in planta. Specific PCR primers and probes for the pathovars savastanoi, nerii and fraxini of P. savastanoi were designed to be used in End Point and Real-Time PCR, both with SYBR Green or TaqMan chemistries. The specificity of all these assays was 100%, as assessed by testing forty-four P. savastanoi strains, belonging to the three pathovars and having different geographical origins. For comparison strains from the pathovars phaseolicola and glycinea of P. savastanoi and bacterial epiphytes from P. savastanoi host plants were also assayed, and all of them tested always negative. The analytical detection limits were about 5 - 0.5 pg of pure genomic DNA and about 102 genome equivalents per reaction. Similar analytical thresholds were achieved in Multiplex Real-Time PCR experiments, even on artificially inoculated olive plants. Here for the first time a complex of PCR-based assays were developed for the simultaneous discrimination and detection of P. savastanoi pv. savastanoi, pv. nerii and pv. fraxini. These tests were shown to be highly reliable, pathovar-specific, sensitive, rapid and able to quantify these pathogens, both in multiplex reactions and in vivo. Compared with the other methods already available for P. savastanoi, the identification

  14. A fiducial detection algorithm for real-time image guided IMRT based on simultaneous MV and kV imaging.

    Science.gov (United States)

    Mao, Weihua; Riaz, Nadeem; Lee, Louis; Wiersma, Rodney; Xing, Lei

    2008-08-01

    The advantage of highly conformal dose techniques such as 3DCRT and IMRT is limited by intrafraction organ motion. A new approach to gain near real-time 3D positions of internally implanted fiducial markers is to analyze simultaneous onboard kV beam and treatment MV beam images (from fluoroscopic or electronic portal image devices). Before we can use this real-time image guidance for clinical 3DCRT and IMRT treatments, four outstanding issues need to be addressed. (1) How will fiducial motion blur the image and hinder tracking fiducials? kV and MV images are acquired while the tumor is moving at various speeds. We find that a fiducial can be successfully detected at a maximum linear speed of 1.6 cm/s. (2) How does MV beam scattering affect kV imaging? We investigate this by varying MV field size and kV source to imager distance, and find that common treatment MV beams do not hinder fiducial detection in simultaneous kV images. (3) How can one detect fiducials on images from 3DCRT and IMRT treatment beams when the MV fields are modified by a multileaf collimator (MLC)? The presented analysis is capable of segmenting a MV field from the blocking MLC and detecting visible fiducials. This enables the calculation of nearly real-time 3D positions of markers during a real treatment. (4) Is the analysis fast enough to track fiducials in nearly real time? Multiple methods are adopted to predict marker positions and reduce search regions. The average detection time per frame for three markers in a 1024 x 768 image was reduced to 0.1 s or less. Solving these four issues paves the way to tracking moving fiducial markers throughout a 3DCRT or IMRT treatment. Altogether, these four studies demonstrate that our algorithm can track fiducials in real time, on degraded kV images (MV scatter), in rapidly moving tumors (fiducial blurring), and even provide useful information in the case when some fiducials are blocked from view by the MLC. This technique can provide a gating signal or

  15. Constructed ILs coated porous magnetic nickel cobaltate hexagonal nanoplates sensing materials for the simultaneous detection of cumulative toxic metals

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Yuanyuan; Zhang, Lei, E-mail: zhanglei63@126.com

    2017-07-05

    Highlights: • A novel sensor material based on ionic liquids@nickel cobaltate was constructed. • Various morphologies of NiCo{sub 2}O{sub 4} were synthesized for electrocatalytic comparison. • ILs@NiCo{sub 2}O{sub 4}-P was used to detect cumulative toxic metals for the first time. • The sensor displayed well reproducibility, excellent selectivity and sensitivity. • The method was applied to detect practical samples with satisfactory results. - Abstract: The different morphologies of magnetic nickel cobaltate (NiCo{sub 2}O{sub 4}) electrocatalysts, consisting of nanoparticles (NiCo{sub 2}O{sub 4}-N), nanoplates (NiCo{sub 2}O{sub 4}-P) and microspheres (NiCo{sub 2}O{sub 4}-S) were fabricated. It was found that the electrocatalytic properties of the sensing materials were strongly dependent on morphology and specific surface area. The porous NiCo{sub 2}O{sub 4} hexagonal nanoplates coupled with ILs as modified materials (ILs@NiCo{sub 2}O{sub 4}-P) for the simultaneous determination of thallium (Tl{sup +}), lead (Pb{sup 2+}) and copper (Cu{sup 2+}), exhibited high sensitivity, long-time stability and good repeatability. The enhanced electrocatalytic activity was attributed to relatively large specific surface area, excellent electronic conductivity, and unique porous nanostructure. The analytical performance of the constructed electrode on detection of Tl{sup +}, Pb{sup 2+} and Cu{sup 2+} was examined using differential pulse anodic stripping voltammetry (DPASV). Under optimal conditions, the electrode showed a good linear response to Tl{sup +}, Pb{sup 2+}and Cu{sup 2+} in the concentration range of 0.1–100.0, 0.1–100.0 and 0.05–100.0 μg/L, respectively. The detection limits (S/N = 3) were 0.046, 0.034 and 0.029 μg/L for Tl{sup +}, Pb{sup 2+} and Cu{sup 2+}, respectively. The fabricated sensor was successfully applied to detect trace Tl{sup +}, Pb{sup 2+} and Cu{sup 2+} in various water and soil samples with satisfactory results. Hence, this work

  16. Reliable detection of fluence anomalies in EPID-based IMRT pretreatment quality assurance using pixel intensity deviations

    International Nuclear Information System (INIS)

    Gordon, J. J.; Gardner, J. K.; Wang, S.; Siebers, J. V.

    2012-01-01

    Purpose: This work uses repeat images of intensity modulated radiation therapy (IMRT) fields to quantify fluence anomalies (i.e., delivery errors) that can be reliably detected in electronic portal images used for IMRT pretreatment quality assurance. Methods: Repeat images of 11 clinical IMRT fields are acquired on a Varian Trilogy linear accelerator at energies of 6 MV and 18 MV. Acquired images are corrected for output variations and registered to minimize the impact of linear accelerator and electronic portal imaging device (EPID) positioning deviations. Detection studies are performed in which rectangular anomalies of various sizes are inserted into the images. The performance of detection strategies based on pixel intensity deviations (PIDs) and gamma indices is evaluated using receiver operating characteristic analysis. Results: Residual differences between registered images are due to interfraction positional deviations of jaws and multileaf collimator leaves, plus imager noise. Positional deviations produce large intensity differences that degrade anomaly detection. Gradient effects are suppressed in PIDs using gradient scaling. Background noise is suppressed using median filtering. In the majority of images, PID-based detection strategies can reliably detect fluence anomalies of ≥5% in ∼1 mm 2 areas and ≥2% in ∼20 mm 2 areas. Conclusions: The ability to detect small dose differences (≤2%) depends strongly on the level of background noise. This in turn depends on the accuracy of image registration, the quality of the reference image, and field properties. The longer term aim of this work is to develop accurate and reliable methods of detecting IMRT delivery errors and variations. The ability to resolve small anomalies will allow the accuracy of advanced treatment techniques, such as image guided, adaptive, and arc therapies, to be quantified.

  17. Simultaneous Determination of Dexamethasone, Ondansetron, Granisetron, Tropisetron, and Azasetron in Infusion Samples by HPLC with DAD Detection

    Directory of Open Access Journals (Sweden)

    Fu-chao Chen

    2017-01-01

    Full Text Available A simple and rapid high-performance liquid chromatography with diode array detector (HPLC-DAD method has been developed and validated for simultaneous quantification of five antiemetic agents in infusion samples: dexamethasone, ondansetron, granisetron, tropisetron, and azasetron. The chromatographic separation was achieved on a Phenomenex C18 column (4.6 mm × 150 mm, 5 μm using acetonitrile-50 mM KH2PO4 buffer-triethylamine (25 : 74 : 1; v/v; pH 4.0. Flow rate was 1.0 mL/min with a column temperature of 30°C. Validation of the method was made in terms of specificity, linearity, accuracy, and intra- and interday precision, as well as quantification and detection limits. The developed method can be used in the laboratory to routinely quantify dexamethasone, ondansetron, granisetron, tropisetron, and azasetron simultaneously and to evaluate the physicochemical stability of referred drugs in mixtures for endovenous use.

  18. A Method to Simultaneously Detect the Current Sensor Fault and Estimate the State of Energy for Batteries in Electric Vehicles.

    Science.gov (United States)

    Xu, Jun; Wang, Jing; Li, Shiying; Cao, Binggang

    2016-08-19

    Recently, State of energy (SOE) has become one of the most fundamental parameters for battery management systems in electric vehicles. However, current information is critical in SOE estimation and current sensor is usually utilized to obtain the latest current information. However, if the current sensor fails, the SOE estimation may be confronted with large error. Therefore, this paper attempts to make the following contributions: Current sensor fault detection and SOE estimation method is realized simultaneously. Through using the proportional integral observer (PIO) based method, the current sensor fault could be accurately estimated. By taking advantage of the accurate estimated current sensor fault, the influence caused by the current sensor fault can be eliminated and compensated. As a result, the results of the SOE estimation will be influenced little by the fault. In addition, the simulation and experimental workbench is established to verify the proposed method. The results indicate that the current sensor fault can be estimated accurately. Simultaneously, the SOE can also be estimated accurately and the estimation error is influenced little by the fault. The maximum SOE estimation error is less than 2%, even though the large current error caused by the current sensor fault still exists.

  19. Simultaneous Detection of Displacement, Rotation Angle, and Contact Pressure Using Sandpaper Molded Elastomer Based Triple Electrode Sensor.

    Science.gov (United States)

    Choi, Eunsuk; Sul, Onejae; Lee, Seung-Beck

    2017-09-06

    In this article, we report on a flexible sensor based on a sandpaper molded elastomer that simultaneously detects planar displacement, rotation angle, and vertical contact pressure. When displacement, rotation, and contact pressure are applied, the contact area between the translating top elastomer electrode and the stationary three bottom electrodes change characteristically depending on the movement, making it possible to distinguish between them. The sandpaper molded undulating surface of the elastomer reduces friction at the contact allowing the sensor not to affect the movement during measurement. The sensor showed a 0.25 mm −1 displacement sensitivity with a ±33 μm accuracy, a 0.027 degree −1 of rotation sensitivity with ~0.95 degree accuracy, and a 4.96 kP −1 of pressure sensitivity. For possible application to joint movement detection, we demonstrated that our sensor effectively detected the up-and-down motion of a human forefinger and the bending and straightening motion of a human arm.

  20. Simultaneous Determination of Four Active Ingredients in Sargentodoxa cuneata by HPLC Coupled with Evaporative Light Scattering Detection

    Directory of Open Access Journals (Sweden)

    Di-Hua Li

    2016-01-01

    Full Text Available A HPLC coupled with evaporative light scattering detection method had been developed for the simultaneous determination of 3,4-dihydroxyphenylethyl alcohol glycoside, salidroside, chlorogenic acid, and liriodendrin in the stem of Sargentodoxa cuneata. With a C18 column, the analysis was performed using acetonitrile and 0.2% formic acid aqueous solution as mobile phase in gradient program at a flow rate of 0.9 mL/min. The optimum drift tube temperature of evaporative light scattering detection was at 105°C with the air flow rate of 2.5 L/min. The calibration curves showed good linearity during the test ranges. This method was validated for limits of detection and quantification, precision, and reproducibility. The recoveries were within the range of 96.39%–104.64%. The relative standard deviations of intraday and interday precision were less than 2.90% and 3.30%, respectively. The developed method can be successfully used to quantify the four analytes in the stem of Sargentodoxa cuneata from various regions in China.

  1. Fiber-Amplifier-Enhanced QEPAS Sensor for Simultaneous Trace Gas Detection of NH3 and H2S

    Directory of Open Access Journals (Sweden)

    Hongpeng Wu

    2015-10-01

    Full Text Available A selective and sensitive quartz enhanced photoacoustic spectroscopy (QEPAS sensor, employing an erbium-doped fiber amplifier (EDFA, and a distributed feedback (DFB laser operating at 1582 nm was demonstrated for simultaneous detection of ammonia (NH3 and hydrogen sulfide (H2S. Two interference-free absorption lines located at 6322.45 cm−1 and 6328.88 cm−1 for NH3 and H2S detection, respectively, were identified. The sensor was optimized in terms of current modulation depth for both of the two target gases. An electrical modulation cancellation unit was equipped to suppress the background noise caused by the stray light. An Allan-Werle variance analysis was performed to investigate the long-term performance of the fiber-amplifier-enhanced QEPAS sensor. Benefitting from the high power boosted by the EDFA, a detection sensitivity (1σ of 52 parts per billion by volume (ppbv and 17 ppbv for NH3 and H2S, respectively, were achieved with a 132 s data acquisition time at atmospheric pressure and room temperature.

  2. Fiber-Amplifier-Enhanced QEPAS Sensor for Simultaneous Trace Gas Detection of NH3 and H2S

    Science.gov (United States)

    Wu, Hongpeng; Dong, Lei; Liu, Xiaoli; Zheng, Huadan; Yin, Xukun; Ma, Weiguang; Zhang, Lei; Yin, Wangbao; Jia, Suotang

    2015-01-01

    A selective and sensitive quartz enhanced photoacoustic spectroscopy (QEPAS) sensor, employing an erbium-doped fiber amplifier (EDFA), and a distributed feedback (DFB) laser operating at 1582 nm was demonstrated for simultaneous detection of ammonia (NH3) and hydrogen sulfide (H2S). Two interference-free absorption lines located at 6322.45 cm−1 and 6328.88 cm−1 for NH3 and H2S detection, respectively, were identified. The sensor was optimized in terms of current modulation depth for both of the two target gases. An electrical modulation cancellation unit was equipped to suppress the background noise caused by the stray light. An Allan-Werle variance analysis was performed to investigate the long-term performance of the fiber-amplifier-enhanced QEPAS sensor. Benefitting from the high power boosted by the EDFA, a detection sensitivity (1σ) of 52 parts per billion by volume (ppbv) and 17 ppbv for NH3 and H2S, respectively, were achieved with a 132 s data acquisition time at atmospheric pressure and room temperature. PMID:26506351

  3. A Multiplex PCR for Simultaneous Detection of Three Zoonotic Parasites Ancylostoma ceylanicum, A. caninum, and Giardia lamblia Assemblage A

    Directory of Open Access Journals (Sweden)

    Wei Hu

    2015-01-01

    Full Text Available Ancylostoma ceylanicum, A. caninum, and Giardia lamblia assemblage A are common intestinal parasites of dogs and cats; they can also infect humans, causing parasitic zoonoses. In this study, a multiplex PCR method was developed for simultaneous identification and detection of those three zoonotic parasites. Three pairs of specific primers were designed based on ITS sequence of A. ceylanicum and A. caninum and TPI gene of G. lamblia available in the GenBank. The multiplex PCR reaction system was established by optimizing the reaction condition, and a series of tests on the sensitivity, specificity, and clinical application were also conducted. Results showed that three target fragments were amplified specifically; the detection limit was 10 eggs for both A. ceylanicum and A. caninum, 72 pg DNA for G. lamblia. Of 112 clinical fecal samples, 34.8% and 17.8% samples were positive for A. caninum and A. ceylanicum, respectively, while only 2.7% samples were positive for G. lamblia assemblage A. It is concluded that the established multiplex PCR assay is a convenient, rapid, cost-effective, and high-efficiency method for molecular detection and epidemiological investigation of three zoonotic parasites.

  4. Sensitive simultaneous detection of seven sexually transmitted agents in semen by multiplex-PCR and of HPV by single PCR.

    Directory of Open Access Journals (Sweden)

    Fabrícia Gimenes

    Full Text Available Sexually transmitted diseases (STDs may impair sperm parameters and functions thereby promoting male infertility. To date limited molecular studies were conducted to evaluate the frequency and type of such infections in semen Thus, we aimed at conceiving and validating a multiplex PCR (M-PCR assay for the simultaneous detection of the following STD pathogens in semen: Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Herpes virus simplex (HSV -1 and -2, and Treponema pallidum; We also investigated the potential usefulness of this M-PCR assay in screening programs for semen pathogens. In addition, we aimed: to detect human Papillomavirus (HPV and genotypes by single PCR (sPCR in the same semen samples; to determine the prevalence of the seven STDs, HPV and co-infections; to assess the possibility that these infections affect semen parameters and thus fertility. The overall validation parameters of M-PCR were extremely high including agreement (99.2%, sensitivity (100.00%, specificity (99.70%, positive (96.40% and negative predictive values (100.00% and accuracy (99.80%. The prevalence of STDs was very high (55.3%. Furthermore, associations were observed between STDs and changes in semen parameters, highlighting the importance of STD detection in semen. Thus, this M-PCR assay has great potential for application in semen screening programs for pathogens in infertility and STD clinics and in sperm banks.

  5. Development of duplex PCR for simultaneous detection of Theileria spp. and Anaplasma spp. in sheep and goats.

    Science.gov (United States)

    Cui, Yanyan; Zhang, Yan; Jian, Fuchun; Zhang, Longxian; Wang, Rongjun; Cao, Shuxuan; Wang, Xiaoxing; Yan, Yaqun; Ning, Changshen

    2017-05-01

    Theileria spp. and Anaplasma spp., which are important tick-borne pathogens (TBPs), impact the health of humans and animals in tropical and subtropical areas. Theileria and Anaplasma co-infections are common in sheep and goats. Following alignment of the relevant DNA sequences, two primer sets were designed to specifically target the Theileria spp. 18S rRNA and Anaplasma spp. 16S rRNA gene sequences. Genomic DNA from the two genera was serially diluted tenfold for testing the sensitivities of detection of the primer sets. The specificities of the primer sets were confirmed when DNA from Anaplasma and Theileria (positive controls), other related hematoparasites (negative controls) and ddH 2 O were used as templates. Fifty field samples were also used to evaluate the utility of single PCR and duplex PCR assays, and the detection results were compared with those of the PCR methods previously published. An optimized duplex PCR assay was established from the two primer sets based on the relevant genes from the two TBPs, and this assay generated products of 298-bp (Theileria spp.) and 139-bp (Anaplasma spp.). The detection limit of the assay was 29.4 × 10 -3  ng per μl, and there was no cross-reaction with the DNA from other hematoparasites. The results showed that the newly developed duplex PCR assay had an efficiency of detection (P > 0.05) similar to other published PCR methods. In this study, a duplex PCR assay was developed that can simultaneously identify Theileria spp. and Anaplasma spp. in sheep and goats. This duplex PCR is a potentially valuable assay for epidemiological studies of TBPs in that it can detect cases of mixed infections of the pathogens. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Facile sysnthesis of fluorescent silver nanoclusters as simultaneous detection and remediation for Hg{sup 2+}

    Energy Technology Data Exchange (ETDEWEB)

    Li, Dan; Li, Biao; Lee, Go Eun; Yang, Sung Ik [Dept. of Applied Chemistry, Kyung Hee University, Yongin (Korea, Republic of)

    2015-06-15

    Mercury is one of the most toxic metals to the environment and human life. 1 This metal can cause serious health problems because it easily passes through skin, respiratory tract, and gastrointestinal tissues into the human body, which will damage kidney, central nervou s system, and endocrine system. A facile synthetic strategy for the preparation of highly fluorescent AgNCs with red emission (λ em = 671 nm, quantum yield = 3.3%) has been developed. AgNCs showed a high sensitivity and selectivity toward the Hg{sup 2+} ions over other metal ions in spiked tap water. The detection limit was determined to be 1.3 ppb, which met the requirement of the USEPA standards for the maximum allowable level of Hg{sup 2+} in drinking water. AgNCs also showed applicability for remediation of Hg{sup 2+} in polluted water with high removal efficiencies. The employment of AgNCs could be used to monitor and remediate Hg{sup 2+} for the water analysis and purification.

  7. Facile sysnthesis of fluorescent silver nanoclusters as simultaneous detection and remediation for Hg2+

    International Nuclear Information System (INIS)

    Li, Dan; Li, Biao; Lee, Go Eun; Yang, Sung Ik

    2015-01-01

    Mercury is one of the most toxic metals to the environment and human life. 1 This metal can cause serious health problems because it easily passes through skin, respiratory tract, and gastrointestinal tissues into the human body, which will damage kidney, central nervou s system, and endocrine system. A facile synthetic strategy for the preparation of highly fluorescent AgNCs with red emission (λ em = 671 nm, quantum yield = 3.3%) has been developed. AgNCs showed a high sensitivity and selectivity toward the Hg 2+ ions over other metal ions in spiked tap water. The detection limit was determined to be 1.3 ppb, which met the requirement of the USEPA standards for the maximum allowable level of Hg 2+ in drinking water. AgNCs also showed applicability for remediation of Hg 2+ in polluted water with high removal efficiencies. The employment of AgNCs could be used to monitor and remediate Hg 2+ for the water analysis and purification.

  8. Development of an ELISA microarray assay for the sensitive and simultaneous detection of ten biodefense toxins.

    Energy Technology Data Exchange (ETDEWEB)

    Jenko, Kathryn; Zhang, Yanfeng; Kostenko, Yulia; Fan, Yongfeng; Garcia-Rodriguez, Consuelo; Lou, Jianlong; Marks, James D.; Varnum, Susan M.

    2014-10-21

    Plant and microbial toxins are considered bioterrorism threat agents because of their extreme toxicity and/or ease of availability. Additionally, some of these toxins are increasingly responsible for accidental food poisonings. The current study utilized an ELISA-based protein antibody microarray for the multiplexed detection of ten biothreat toxins, botulinum neurotoxins (BoNT) A, B, C, D, E, F, ricin, shiga toxins 1 and 2 (Stx), and staphylococcus enterotoxin B (SEB), in buffer and complex biological matrices. The multiplexed assay displayed a sensitivity of 1.3 pg/mL (BoNT/A, BoNT/B, SEB, Stx-1 and Stx-2), 3.3 pg/mL (BoNT/C, BoNT/E, BoNT/F) and 8.2 pg/mL (BoNT/D, ricin). All assays demonstrated high accuracy (75-120 percent recovery) and reproducibility (most coefficients of variation < 20%). Quantification curves for the ten toxins were also evaluated in clinical samples (serum, plasma, nasal fluid, saliva, stool, and urine) and environmental samples (apple juice, milk and baby food) with overall minimal matrix effects. The multiplex assays were highly specific, with little crossreactivity observed between the selected toxin antibodies. The results demonstrate a multiplex microarray that improves current immunoassay sensitivity for biological warfare agents in buffer, clinical, and environmental samples.

  9. Simultaneous detection of four garlic viruses by multiplex reverse transcription PCR and their distribution in Indian garlic accessions.

    Science.gov (United States)

    Majumder, S; Baranwal, V K

    2014-06-01

    Indian garlic is infected with Onion yellow dwarf virus (OYDV), Shallot latent virus (SLV), Garlic common latent virus (GarCLV) and allexiviruses. Identity and distribution of garlic viruses in various garlic accessions from different geographical regions of India were investigated. OYDV and allexiviruses were observed in all the garlic accessions, while SLV and GarCLV were observed only in a few accessions. A multiplex reverse transcription (RT)-PCR method was developed for the simultaneous detection and identification of OYDV, SLV, GarCLV and Allexivirus infecting garlic accessions in India. This multiplex protocol standardized in this study will be useful in indexing of garlic viruses and production of virus free seed material. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. infrared spectrophotometer for simultaneous detection of traces of heavy water and indocyanine green in flowing blood. In vivo experimentation

    International Nuclear Information System (INIS)

    Capitini, R.; Roveyaz, P.

    1981-07-01

    We have developed an infrared absorption method for the pulmonary extravascular water and cardiac output determination by the multiple indicator technique. This led us to construct an original double differential spectrophotometer (DUPLEX) for simultaneous detection of traces of heavy water (D 2 O) and indocyanine green (ICG) in circulating blood in a single absorption cell. This DUPLEX is connected to a computer and results are treated on line. D 2 O and ICG are used as non toxic diffusible and vascular tracers respectively. Performances of the DUPLEX are given and show a high accuracy with D 2 O (x 10) and ICG (x 2) compared with the commercial optical analysers. We show the validity of the method for determining the cardiac output and the pulmonary extravascular water in the course of numerous experiments on human and rats subjects. Preliminary results concerning the measurement of global water on rats are also given [fr

  11. Point-of-need simultaneous electrochemical detection of lead and cadmium using low-cost stencil-printed transparency electrodes.

    Science.gov (United States)

    Martín-Yerga, Daniel; Álvarez-Martos, Isabel; Blanco-López, M Carmen; Henry, Charles S; Fernández-Abedul, M Teresa

    2017-08-15

    In this work, we report a simple and yet efficient stencil-printed electrochemical platform that can be integrated into the caps of sample containers and thus, allows in-field quantification of Cd(II) and Pb(II) in river water samples. The device exploits the low-cost features of carbon (as electrode material) and paper/polyester transparency sheets (as substrate). Electrochemical analysis of the working electrodes prepared on different substrates (polyester transparency sheets, chromatographic, tracing and office papers) with hexaammineruthenium(III) showed that their electroactive area and electron transfer kinetics are highly affected by the porosity of the material. Electrodes prepared on transparency substrates showed the best electroanalytical performance for the simultaneous determination of Cd(II) and Pb(II) by square-wave anodic stripping voltammetry. Interestingly, the temperature and time at which the carbon ink was cured had significant effect on the electrochemical response, especially the capacitive current. The amount of Cd and Pb on the electrode surface can be increased about 20% by in situ electrodeposition of bismuth. The electrochemical platform showed a linear range comprised between 1 and 200 μg/L for both metals, sensitivity of analysis of 0.22 and 0.087 μA/ppb and limits of detection of 0.2 and 0.3 μg/L for Cd(II) and Pb(II), respectively. The analysis of river water samples was done directly in the container where the sample was collected, which simplifies the procedure and approaches field analysis. The developed point-of-need detection system allowed simultaneous determination of Cd(II) and Pb(II) in those samples using the standard addition method with precise and accurate results. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Simultaneous Electrochemical Detection of Dopamine and Ascorbic Acid Using an Iron Oxide/Reduced Graphene Oxide Modified Glassy Carbon Electrode

    Directory of Open Access Journals (Sweden)

    Teo Peik-See

    2014-08-01

    Full Text Available The fabrication of an electrochemical sensor based on an iron oxide/graphene modified glassy carbon electrode (Fe3O4/rGO/GCE and its simultaneous detection of dopamine (DA and ascorbic acid (AA is described here. The Fe3O4/rGO nanocomposite was synthesized via a simple, one step in-situ wet chemical method and characterized by different techniques. The presence of Fe3O4 nanoparticles on the surface of rGO sheets was confirmed by FESEM and TEM images. The electrochemical behavior of Fe3O4/rGO/GCE towards electrocatalytic oxidation of DA was investigated by cyclic voltammetry (CV and differential pulse voltammetry (DPV analysis. The electrochemical studies revealed that the Fe3O4/rGO/GCE dramatically increased the current response against the DA, due to the synergistic effect emerged between Fe3O4 and rGO. This implies that Fe3O4/rGO/GCE could exhibit excellent electrocatalytic activity and remarkable electron transfer kinetics towards the oxidation of DA. Moreover, the modified sensor electrode portrayed sensitivity and selectivity for simultaneous determination of AA and DA. The observed DPVs response linearly depends on AA and DA concentration in the range of 1–9 mM and 0.5–100 µM, with correlation coefficients of 0.995 and 0.996, respectively. The detection limit of (S/N = 3 was found to be 0.42 and 0.12 µM for AA and DA, respectively.

  13. Simultaneous determination of sulpiride and mebeverine by HPLC method using fluorescence detection: application to real human plasma

    Directory of Open Access Journals (Sweden)

    Walash Mohamed I

    2012-02-01

    Full Text Available Abstract A new simple, rapid and sensitive reversed-phase liquid chromatographic method was developed and validated for the simultaneous determination of sulpiride (SUL and mebeverine Hydrochloride (MEB in the presence of their impurities and degradation products. The separation of these compounds was achieved within 6 min on a 250 mm, 4.6 mm i.d., 5 m particle size Waters®-C18 column using isocractic mobile phase containing a mixture of acetonitrile and 0.01 M dihydrogenphosphate buffer (45:55 at pH = 4.0. The analysis was performed at a flow rate of 1.0 mL/min with fluorescence-detection at excitation 300 nm and emission at 365 nm. The concentration-response relationship was linear over a concentration range of 10- 100 ng/mL for both MEB and SUL with a limit of detection 0.73 ng/mL and 0.85 ng/mL for MEB and SUL respectively. The proposed method was successfully applied for the analysis of both MEB and SUL in bulk with average recoveries of 100.22 ± 0.757% and 99.96 ± 0.625% respectively, and in commercial tablets with average recoveries of 100.04 ± 0.93% and 100.03 ± 0.376% for MEB and SUL respectively. The proposed method was successfully applied to the determination of MEB metabolite (veratic acid in real plasma simultaneously with SUL. The mean% recoveries (n = 3 for both MEB metabolite (veratic acid and SUL were 100.36 ± 2.92 and 99.06 ± 2.11 for spiked human plasma respectively. For real human plasma, the mean% recoveries (n = 3 were and respectively.

  14. Gas chromatography with simultaneous detection: Ultraviolet spectroscopy, flame ionization, and mass spectrometry.

    Science.gov (United States)

    Gras, Ronda; Luong, Jim; Haddad, Paul R; Shellie, Robert A

    2018-05-08

    An effective analytical strategy was developed and implemented to exploit the synergy derived from three different detector classes for gas chromatography, namely ultraviolet spectroscopy, flame ionization, and mass spectrometry for volatile compound analysis. This strategy was achieved by successfully hyphenating a user-selectable multi-wavelength diode array detector featuring a positive temperature coefficient thermistor as an isothermal heater to a gas chromatograph. By exploiting the non-destructive nature of the diode array detector, the effluent from the detector was split to two parallel detectors; namely a quadrupole mass spectrometer and a flame ionization detector. This multi-hyphenated configuration with the use of three detectors is a powerful approach not only for selective detection enhancement but also for improvement in structural elucidation of volatile compounds where fewer fragments can be obtained or for isomeric compound analysis. With the diode array detector capable of generating high resolution gas phase spectra, the information collected provides useful confirmatory information without a total dependence on the chromatographic separation process which is based on retention time. This information-rich approach to chromatography is achieved without incurring extra analytical time, resulting in improvements in compound identification accuracy, analytical productivity, and cost. Chromatographic performance obtained from model compounds was found to be acceptable with a relative standard deviation of the retention times of less than 0.01% RSD, and a repeatability at two levels of concentration of 100 and 1000 ppm (v/v) of less than 5% (n = 10). With this configuration, correlation of data between the three detectors was simplified by having near identical retention times for the analytes studied. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Realization of ppm-level CO detection with exceptionally high sensitivity using reduced graphene oxide-loaded SnO2 nanofibers with simultaneous Au functionalization.

    Science.gov (United States)

    Kim, Jae-Hun; Katoch, Akash; Kim, Hyoun Woo; Kim, Sang Sub

    2016-03-07

    We have realized the highly sensitive, selective ppm-level carbon monoxide (CO) detection based on graphene oxide (RGO) nanosheets-loaded SnO2 nanofibers with simultaneous Au functionalization. The interplay between RGO/Au and SnO2 in terms of transfer of charge carriers and modulation of potential barriers is responsible for the exceptionally high CO detectability.

  16. Simultaneous detection of dual biomarkers from humans exposed to organophosphorus pesticides by combination of immunochromatographic test strip and ellman assay.

    Science.gov (United States)

    Yang, Mingming; Zhao, Yuting; Wang, Limin; Paulsen, Michael; Simpson, Christopher D; Liu, Fengquan; Du, Dan; Lin, Yuehe

    2018-05-01

    A novel sandwich immunoassay based immunochromatographic test strip (ICTS) has been developed for simultaneously measuring both butyrylcholinesterase (BChE) activity and the total amount of BChE (including inhibited and active enzyme) from 70 μLpost-exposure human plasma sample. The principle of this method is based on the BChE monoclonal antibody (MAb) capable of acting as both capture antibody and detection antibody. The BChE MAb which was immobilized on the test line was able to recognize both organophosphorus BChE adducts (OP-BChE) and BChE and provided equal binding affinity, permitting detection of the total enzyme amount in post-exposure human plasma samples. The formed immunocomplexes on the test line can further be excised from the test-strip for subsequent off-line measurement of BChE activity using the Ellman assay. Therefore, dual biomarkers of BChE activity and phosphorylation (OP-BChE) will be obtained simultaneously. The whole sandwich-immunoassay was performed on one ICTS, greatly reducing analytical time. The ICTS sensor showed excellent linear responses for assaying total amount of BChE and active BChE ranging from 0.22 to 3.58nM and 0.22-7.17nM, respectively. Both the signal detection limits are 0.10nM. We validated the practical application of the proposed method to measure 124 human plasma samples from orchard workers and cotton farmers with long-term exposure to organophosphorus pesticides (OPs). The results were in highly agreement with LC/MS/MS which verified our method is extremely accurate. Combining the portability and rapidity of test strip and the compatibility of BChE MAb as both capture antibody and detection antibody, the developed method provides a baseline-free, low-cost and rapid tool for in-field monitoring of OP exposures. Copyright © 2018 Elsevier B.V. All rights reserved.

  17. Multiplex Nucleic Acid Sequence-Based Amplification for Simultaneous Detection of Several Enteric Viruses in Model Ready-To-Eat Foods†

    Science.gov (United States)

    Jean, Julie; D'Souza, Doris H.; Jaykus, Lee-Ann

    2004-01-01

    Human enteric viruses are currently recognized as one of the most important causes of food-borne disease. Implication of enteric viruses in food-borne outbreaks can be difficult to confirm due to the inadequacy of the detection methods available. In this study, a nucleic acid sequence-based amplification (NASBA) method was developed in a multiplex format for the specific, simultaneous, and rapid detection of epidemiologically relevant human enteric viruses. Three previously reported primer sets were used in a single reaction for the amplification of RNA target fragments of 474, 371, and 165 nucleotides for the detection of hepatitis A virus and genogroup I and genogroup II noroviruses, respectively. Amplicons were detected by agarose gel electrophoresis and confirmed by electrochemiluminescence and Northern hybridization. Endpoint detection sensitivity for the multiplex NASBA assay was approximately 10−1 reverse transcription-PCR-detectable units (or PFU, as appropriate) per reaction. When representative ready-to-eat foods (deli sliced turkey and lettuce) were inoculated with various concentrations of each virus and processed for virus detection with the multiplex NASBA method, all three human enteric viruses were simultaneously detected at initial inoculum levels of 100 to 102 reverse transcription-PCR-detectable units (or PFU)/9 cm2 in both food commodities. The multiplex NASBA system provides rapid and simultaneous detection of clinically relevant food-borne viruses in a single reaction tube and may be a promising alternative to reverse transcription-PCR for the detection of viral contamination of foods. PMID:15528524

  18. Multiplex nucleic acid sequence-based amplification for simultaneous detection of several enteric viruses in model ready-to-eat foods.

    Science.gov (United States)

    Jean, Julie; D'Souza, Doris H; Jaykus, Lee-Ann

    2004-11-01

    Human enteric viruses are currently recognized as one of the most important causes of food-borne disease. Implication of enteric viruses in food-borne outbreaks can be difficult to confirm due to the inadequacy of the detection methods available. In this study, a nucleic acid sequence-based amplification (NASBA) method was developed in a multiplex format for the specific, simultaneous, and rapid detection of epidemiologically relevant human enteric viruses. Three previously reported primer sets were used in a single reaction for the amplification of RNA target fragments of 474, 371, and 165 nucleotides for the detection of hepatitis A virus and genogroup I and genogroup II noroviruses, respectively. Amplicons were detected by agarose gel electrophoresis and confirmed by electrochemiluminescence and Northern hybridization. Endpoint detection sensitivity for the multiplex NASBA assay was approximately 10(-1) reverse transcription-PCR-detectable units (or PFU, as appropriate) per reaction. When representative ready-to-eat foods (deli sliced turkey and lettuce) were inoculated with various concentrations of each virus and processed for virus detection with the multiplex NASBA method, all three human enteric viruses were simultaneously detected at initial inoculum levels of 10(0) to 10(2) reverse transcription-PCR-detectable units (or PFU)/9 cm2 in both food commodities. The multiplex NASBA system provides rapid and simultaneous detection of clinically relevant food-borne viruses in a single reaction tube and may be a promising alternative to reverse transcription-PCR for the detection of viral contamination of foods.

  19. Simultaneous determination of piracetam and its four impurities by RP-HPLC with UV detection.

    Science.gov (United States)

    Arayne, M Saeed; Sultana, Najma; Siddiqui, Farhan Ahmed; Mirza, Agha Zeeshan; Qureshi, Faiza; Zuberi, M Hashim

    2010-08-01

    A simple and rapid high-performance liquid chromatographic method for the separation and determination of piracetam and its four impurities, 2-oxopyrrolidin-1-yl)acetic acid, pyrrolidin-2-one, methyl (2-oxopyrrolidin-1-yl)acetate, and ethyl (2-oxopyrrolidin-1-yl)acetate, was developed. The separation was achieved on a reversed-phase C(18) Nucleosil column (25 cm x 0.46 cm, 10 microm). The mobile phase is composed of an aqueous solution containing 0.2 g/L of triethyl amine-acetonitrile (85:15, v/v). The pH of the mobile phase was adjusted to 6.5 with phosphoric acid at a flow rate of 1 mL/min at ambient temperature and UV detection at 205 nm. The developed method was found to give good separation between the pure drug and its four related substance. The polynomial regression data for the calibration plots showed good linear relationship in the concentration range of 50-10,000 ng/mL, 25-10,000 ng/mL, 45-10,000 ng/mL, 34-10,000 ng/mL, and 55-10,000 ng/mL, respectively, with r(2) = 0.9999. The method was validated for precision, accuracy, ruggedness, and recovery. The minimum quantifiable amounts were found to be 50 ng/mL of piracetam, 25 ng/mL of 2-oxopyrrolidin-1-yl)acetic acid, 45 ng/mL of pyrrolidin-2-one, 34 ng/mL of methyl (2-oxopyrrolidin-1-yl)acetate, and 55 ng/mL of ethyl (2-oxopyrrolidin-1-yl)acetate. Statistical analysis proves that the method is reproducible and selective for the estimation of piracetam as well as its related substance. As the method could effectively separate the drug from the related substances, it can be employed as a stability-indicating one. The proposed method shows high efficiency, allowing the separation of the main component piracetam from other impurities.

  20. A Type-2 fuzzy data fusion approach for building reliable weighted protein interaction networks with application in protein complex detection.

    Science.gov (United States)

    Mehranfar, Adele; Ghadiri, Nasser; Kouhsar, Morteza; Golshani, Ashkan

    2017-09-01

    Detecting the protein complexes is an important task in analyzing the protein interaction networks. Although many algorithms predict protein complexes in different ways, surveys on the interaction networks indicate that about 50% of detected interactions are false positives. Consequently, the accuracy of existing methods needs to be improved. In this paper we propose a novel algorithm to detect the protein complexes in 'noisy' protein interaction data. First, we integrate several biological data sources to determine the reliability of each interaction and determine more accurate weights for the interactions. A data fusion component is used for this step, based on the interval type-2 fuzzy voter that provides an efficient combination of the information sources. This fusion component detects the errors and diminishes their effect on the detection protein complexes. So in the first step, the reliability scores have been assigned for every interaction in the network. In the second step, we have proposed a general protein complex detection algorithm by exploiting and adopting the strong points of other algorithms and existing hypotheses regarding real complexes. Finally, the proposed method has been applied for the yeast interaction datasets for predicting the interactions. The results show that our framework has a better performance regarding precision and F-measure than the existing approaches. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Simultaneous detection and identification of Aspergillus and mucorales species in tissues collected from patients with fungal rhinosinusitis.

    Science.gov (United States)

    Zhao, Zuotao; Li, Lili; Wan, Zhe; Chen, Wei; Liu, Honggang; Li, Ruoyu

    2011-04-01

    Rapid detection and differentiation of Aspergillus and Mucorales species in fungal rhinosinusitis diagnosis are desirable, since the clinical management and prognosis associated with the two taxa are fundamentally different. We describe an assay based on a combination of broad-range PCR amplification and reverse line blot hybridization (PCR/RLB) to detect and differentiate the pathogens causing fungal rhinosinusitis, which include five Aspergillus species (A. fumigatus, A. flavus, A. niger, A. terreus, and A. nidulans) and seven Mucorales species (Mucor heimalis, Mucor racemosus, Mucor cercinelloidea, Rhizopus arrhizus, Rhizopus microsporus, Rhizomucor pusillus, and Absidia corymbifera). The assay was validated with 98 well-characterized clinical isolates and 41 clinical tissue specimens. PCR/RLB showed high sensitivity and specificity, with 100% correct identifications of 98 clinical isolates and no cross-hybridization between the species-specific probes. Results for five control isolates, Candida albicans, Fusarium solani, Scedosporium apiospermum, Penicillium marneffei, and Exophiala verrucosa, were negative as judged by PCR/RLB. The analytical sensitivity of PCR/RLB was found to be 1.8 × 10(-3) ng/μl by 10-fold serial dilution of Aspergillus genomic DNA. The assay identified 35 of 41 (85.4%) clinical specimens, exhibiting a higher sensitivity than fungal culture (22 of 41; 53.7%) and direct sequencing (18 of 41; 43.9%). PCR/RLB similarly showed high specificity, with correct identification 16 of 18 specimens detected by internal transcribed spacer (ITS) sequencing and 16 of 22 detected by fungal culture, but it also has the additional advantage of being able to detect mixed infection in a single clinical specimen. The PCR/RLB assay thus provides a rapid and reliable option for laboratory diagnosis of fungal rhinosinusitis.

  2. Simultaneous Detection and Identification of Aspergillus and Mucorales Species in Tissues Collected from Patients with Fungal Rhinosinusitis▿

    Science.gov (United States)

    Zhao, Zuotao; Li, Lili; Wan, Zhe; Chen, Wei; Liu, Honggang; Li, Ruoyu

    2011-01-01

    Rapid detection and differentiation of Aspergillus and Mucorales species in fungal rhinosinusitis diagnosis are desirable, since the clinical management and prognosis associated with the two taxa are fundamentally different. We describe an assay based on a combination of broad-range PCR amplification and reverse line blot hybridization (PCR/RLB) to detect and differentiate the pathogens causing fungal rhinosinusitis, which include five Aspergillus species (A. fumigatus, A. flavus, A. niger, A. terreus, and A. nidulans) and seven Mucorales species (Mucor heimalis, Mucor racemosus, Mucor cercinelloidea, Rhizopus arrhizus, Rhizopus microsporus, Rhizomucor pusillus, and Absidia corymbifera). The assay was validated with 98 well-characterized clinical isolates and 41 clinical tissue specimens. PCR/RLB showed high sensitivity and specificity, with 100% correct identifications of 98 clinical isolates and no cross-hybridization between the species-specific probes. Results for five control isolates, Candida albicans, Fusarium solani, Scedosporium apiospermum, Penicillium marneffei, and Exophiala verrucosa, were negative as judged by PCR/RLB. The analytical sensitivity of PCR/RLB was found to be 1.8 × 10−3 ng/μl by 10-fold serial dilution of Aspergillus genomic DNA. The assay identified 35 of 41 (85.4%) clinical specimens, exhibiting a higher sensitivity than fungal culture (22 of 41; 53.7%) and direct sequencing (18 of 41; 43.9%). PCR/RLB similarly showed high specificity, with correct identification 16 of 18 specimens detected by internal transcribed spacer (ITS) sequencing and 16 of 22 detected by fungal culture, but it also has the additional advantage of being able to detect mixed infection in a single clinical specimen. The PCR/RLB assay thus provides a rapid and reliable option for laboratory diagnosis of fungal rhinosinusitis. PMID:21325541

  3. Novel algorithm for simultaneous component detection and pseudo-molecular ion characterization in liquid chromatography–mass spectrometry

    International Nuclear Information System (INIS)

    Zhang, Yufeng; Wang, Xiaoan; Wo, Siukwan; Ho, Hingman; Han, Quanbin; Fan, Xiaohui; Zuo, Zhong

    2015-01-01

    Highlights: • Novel stepwise component detection algorithm (SCDA) for LC–MS datasets. • New isotopic distribution and adduct-ion models for mass spectra. • Automatic component classification based on adduct-ion and isotopic distributions. - Abstract: Resolving components and determining their pseudo-molecular ions (PMIs) are crucial steps in identifying complex herbal mixtures by liquid chromatography–mass spectrometry. To tackle such labor-intensive steps, we present here a novel algorithm for simultaneous detection of components and their PMIs. Our method consists of three steps: (1) obtaining a simplified dataset containing only mono-isotopic masses by removal of background noise and isotopic cluster ions based on the isotopic distribution model derived from all the reported natural compounds in dictionary of natural products; (2) stepwise resolving and removing all features of the highest abundant component from current simplified dataset and calculating PMI of each component according to an adduct-ion model, in which all non-fragment ions in a mass spectrum are considered as PMI plus one or several neutral species; (3) visual classification of detected components by principal component analysis (PCA) to exclude possible non-natural compounds (such as pharmaceutical excipients). This algorithm has been successfully applied to a standard mixture and three herbal extract/preparations. It indicated that our algorithm could detect components’ features as a whole and report their PMI with an accuracy of more than 98%. Furthermore, components originated from excipients/contaminants could be easily separated from those natural components in the bi-plots of PCA

  4. Novel algorithm for simultaneous component detection and pseudo-molecular ion characterization in liquid chromatography–mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yufeng; Wang, Xiaoan; Wo, Siukwan [School of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong (China); Ho, Hingman; Han, Quanbin [School of Chinese Medicine, Hong Kong Baptist University, 7 Baptist University Road, Kowloon Tong, Hong Kong (China); Fan, Xiaohui [College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058 (China); Zuo, Zhong, E-mail: joanzuo@cuhk.edu.hk [School of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong (China)

    2015-01-01

    Highlights: • Novel stepwise component detection algorithm (SCDA) for LC–MS datasets. • New isotopic distribution and adduct-ion models for mass spectra. • Automatic component classification based on adduct-ion and isotopic distributions. - Abstract: Resolving components and determining their pseudo-molecular ions (PMIs) are crucial steps in identifying complex herbal mixtures by liquid chromatography–mass spectrometry. To tackle such labor-intensive steps, we present here a novel algorithm for simultaneous detection of components and their PMIs. Our method consists of three steps: (1) obtaining a simplified dataset containing only mono-isotopic masses by removal of background noise and isotopic cluster ions based on the isotopic distribution model derived from all the reported natural compounds in dictionary of natural products; (2) stepwise resolving and removing all features of the highest abundant component from current simplified dataset and calculating PMI of each component according to an adduct-ion model, in which all non-fragment ions in a mass spectrum are considered as PMI plus one or several neutral species; (3) visual classification of detected components by principal component analysis (PCA) to exclude possible non-natural compounds (such as pharmaceutical excipients). This algorithm has been successfully applied to a standard mixture and three herbal extract/preparations. It indicated that our algorithm could detect components’ features as a whole and report their PMI with an accuracy of more than 98%. Furthermore, components originated from excipients/contaminants could be easily separated from those natural components in the bi-plots of PCA.

  5. Simultaneous Determination of 14 Phenolic Compounds in Grape Canes by HPLC-DAD-UV Using Wavelength Switching Detection

    Directory of Open Access Journals (Sweden)

    Ang Zhang

    2013-11-01

    Full Text Available The paper described a novel chromatographic method for the simultaneous determination of phenolic compounds such as gallic, protocatechuic, vanillic, caffeic, syringic, p-coumaric and salicylic acid, (+-catechin, (‒-epicatechin, rutin, morin, quercetin, coumarin and trans-resveratrol at their maximum absorbance wavelengths (MAW employing reverse-phase high performance liquid chromatography combined with DAD and UV detection via detection wavelength switching. The method was based on MAW acquisition by DAD and quantification by UV. The separation process was performed on a Shim-Pack VP-ODS C18 column (250 mm × 4.6 mm, 5 μm held at 30 °C, utilizing 3.0% acetic acid and acetonitrile as mobile phase at a flow rate of 0.8 mL/min in the gradient elution mode. The method was fully validated in terms of linearity (r2 > 0.9990, 10‒350 mg/L, precision (both intra-day and inter-day RSD < 4.22%, accuracy (97.31%‒104.66%, specificity, robustness (0.59% < RSD < 2.86%, limit of detection and quantification. The switching method significantly improved the sensitivities of most phenolics studied in comparison with the standard constant wavelength detection (280 nm. The proposed method has been successfully applied to the determination of 14 phenolic compounds in 89 varieties of one-year-old Chinese grape one-year-canes. Grape canes contain many phenolics, especially trans-resveratrol, (‒-epicatechin, and (+-catechin.

  6. Simultaneous determination of 1- and 2-naphthol in human urine using on-line clean-up column-switching liquid chromatography-fluorescence detection.

    Science.gov (United States)

    Preuss, Ralf; Angerer, Jürgen

    2004-03-05

    We developed a new 3-D HPLC method for on-line clean-up and simultaneous quantification of two important naphthalene metabolites, 1-naphthol and 2-naphthol, in human urine. Except an enzymatic hydrolysis no further sample pre-treatment is necessary. The metabolites are stripped from urinary matrix by on-line extraction on a restricted access material pre-column (RAM RP-8), transferred in backflush mode onto a silica-based CN-(cyano)phase column for further purification from interfering substances. By another successive column switching step both analytes are transferred with a minimum of overlapping interferences onto a C12 bonded reversed phase column with trimethylsilyl endcapping where the final separation is carried out. The entire arrangement is software controlled. Eluting analytes are quantified by fluorescence detection (227/430 nm) after an external calibration. Within a total run time of 40 min we can selectively quantify both naphthols with detection limits in the lower ppb range (1.5 and 0.5 microg/l for 1- and 2-naphthol, respectively) with excellent reliability (ensured by precision, accuracy, matrix-independency and FIOH quality assurance program participation). First results on a collective of 53 occupationally non exposed subjects showed mean levels of 11.0 microg/l (1-naphthol) and 12.9 microg/l (2-naphthol). Among smokers (n=21) a significantly elevated mean level of urinary naphthols was determined (1-naphthol: 19.2 microg/l and 2-naphthol: 23.7 microg/l) in comparison to non smokers (n=32; 1-naphthol: 5.6 microg/l, 2-naphthol: 5.6 microg/l).

  7. Electrochemical fecal pellet sensor for simultaneous real-time ex vivo detection of colonic serotonin signalling and motility

    Science.gov (United States)

    Morris, Rachel; Fagan-Murphy, Aidan; MacEachern, Sarah J.; Covill, Derek; Patel, Bhavik Anil

    2016-03-01

    Various investigations have focused on understanding the relationship between mucosal serotonin (5-HT) and colonic motility, however contradictory studies have questioned the importance of this intestinal transmitter. Here we described the fabrication and use of a fecal pellet electrochemical sensor that can be used to simultaneously detect the release of luminal 5-HT and colonic motility. Fecal pellet sensor devices were fabricated using carbon nanotube composite electrodes that were housed in 3D printed components in order to generate a device that had shape and size that mimicked a natural fecal pellet. Devices were fabricated where varying regions of the pellet contained the electrode. Devices showed that they were stable and sensitive for ex vivo detection of 5-HT, and no differences in the fecal pellet velocity was observed when compared to natural fecal pellets. The onset of mucosal 5-HT was observed prior to the movement of the fecal pellet. The release of mucosal 5-HT occurred oral to the fecal pellet and was linked to the contraction of the bowel wall that drove pellet propulsion. Taken, together these findings provide new insights into the role of mucosal 5-HT and suggest that the transmitter acts as a key initiator of fecal pellet propulsion.

  8. The effects of changes in object location on object identity detection: A simultaneous EEG-fMRI study.

    Science.gov (United States)

    Yang, Ping; Fan, Chenggui; Wang, Min; Fogelson, Noa; Li, Ling

    2017-08-15

    Object identity and location are bound together to form a unique integration that is maintained and processed in visual working memory (VWM). Changes in task-irrelevant object location have been shown to impair the retrieval of memorial representations and the detection of object identity changes. However, the neural correlates of this cognitive process remain largely unknown. In the present study, we aim to investigate the underlying brain activation during object color change detection and the modulatory effects of changes in object location and VWM load. To this end we used simultaneous electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) recordings, which can reveal the neural activity with both high temporal and high spatial resolution. Subjects responded faster and with greater accuracy in the repeated compared to the changed object location condition, when a higher VWM load was utilized. These results support the spatial congruency advantage theory and suggest that it is more pronounced with higher VWM load. Furthermore, the spatial congruency effect was associated with larger posterior N1 activity, greater activation of the right inferior frontal gyrus (IFG) and less suppression of the right supramarginal gyrus (SMG), when object location was repeated compared to when it was changed. The ERP-fMRI integrative analysis demonstrated that the object location discrimination-related N1 component is generated in the right SMG. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Simultaneous point-of-care detection of anemia and sickle cell disease in Tanzania: the RAPID study.

    Science.gov (United States)

    Smart, Luke R; Ambrose, Emmanuela E; Raphael, Kevin C; Hokororo, Adolfine; Kamugisha, Erasmus; Tyburski, Erika A; Lam, Wilbur A; Ware, Russell E; McGann, Patrick T

    2018-02-01

    Both anemia and sickle cell disease (SCD) are highly prevalent across sub-Saharan Africa, and limited resources exist to diagnose these conditions quickly and accurately. The development of simple, inexpensive, and accurate point-of-care (POC) assays represents an important advance for global hematology, one that could facilitate timely and life-saving medical interventions. In this prospective study, Robust Assays for Point-of-care Identification of Disease (RAPID), we simultaneously evaluated a POC immunoassay (Sickle SCAN™) to diagnose SCD and a first-generation POC color-based assay to detect anemia. Performed at Bugando Medical Center in Mwanza, Tanzania, RAPID tested 752 participants (age 1 day to 20 years) in four busy clinical locations. With minimally trained medical staff, the SCD POC assay diagnosed SCD with 98.1% sensitivity and 91.1% specificity. The hemoglobin POC assay had 83.2% sensitivity and 74.5% specificity for detection of severe anemia (Hb ≤ 7 g/dL). Interobserver agreement was excellent for both POC assays (r = 0.95-0.96). Results for the hemoglobin POC assay have informed the second-generation assay design to be more suitable for low-resource settings. RAPID provides practical feasibility data regarding two novel POC assays for the diagnosis of anemia and SCD in real-world field evaluations and documents the utility and potential impact of these POC assays for sub-Saharan Africa.

  10. Double-label immunofluorescence method for simultaneous detection of adenovirus and herpes simplex virus from the eye.

    Science.gov (United States)

    Walpita, P; Darougar, S

    1989-07-01

    The development and application of a double-label immunofluorescence method which has the potential to screen for single or dual infections from any site, in single shell vial cultures, is described. In this study, a total of 1,141 ocular specimens were inoculated in shell vials, centrifuged at 15,000 X g for 1 h, incubated at 37 degrees C for 48 h, and fixed in methanol at room temperature for 15 min. The virus inclusions were detected by staining with a double-label indirect immunofluorescence procedure using mixtures of appropriate first antibodies, followed by fluorescein- and rhodamine-conjugated second antibodies. Each specimen was also inoculated in parallel by the conventional virus isolation method. The sensitivity and specificity of the double-label shell vial procedure were comparable to those with the conventional method, and the former test took only 48 h to complete. The test offers a rapid and simple single-vial procedure which allows for individual or simultaneous detection of multiple pathogens. It results in savings in time and cost over the conventional virus isolation method and other shell vial procedures.

  11. Simultaneous determination of caffeine, paracetamol, and ibuprofen in pharmaceutical formulations by high-performance liquid chromatography with UV detection and by capillary electrophoresis with conductivity detection.

    Science.gov (United States)

    Cunha, Rafael R; Chaves, Sandro C; Ribeiro, Michelle M A C; Torres, Lívia M F C; Muñoz, Rodrigo A A; Dos Santos, Wallans T P; Richter, Eduardo M

    2015-05-01

    Paracetamol, caffeine and ibuprofen are found in over-the-counter pharmaceutical formulations. In this work, we propose two new methods for simultaneous determination of paracetamol, caffeine and ibuprofen in pharmaceutical formulations. One method is based on high-performance liquid chromatography with diode-array detection and the other on capillary electrophoresis with capacitively coupled contactless conductivity detection. The separation by high-performance liquid chromatography with diode-array detection was achieved on a C18 column (250×4.6 mm(2), 5 μm) with a gradient mobile phase comprising 20-100% acetonitrile in 40 mmol L(-1) phosphate buffer pH 7.0. The separation by capillary electrophoresis with capacitively coupled contactless conductivity detection was achieved on a fused-silica capillary (40 cm length, 50 μm i.d.) using 10 mmol L(-1) 3,4-dimethoxycinnamate and 10 mmol L(-1) β-alanine with pH adjustment to 10.4 with lithium hydroxide as background electrolyte. The determination of all three pharmaceuticals was carried out in 9.6 min by liquid chromatography and in 2.2 min by capillary electrophoresis. Detection limits for caffeine, paracetamol and ibuprofen were 4.4, 0.7, and 3.4 μmol L(-1) by liquid chromatography and 39, 32, and 49 μmol L(-1) by capillary electrophoresis, respectively. Recovery values for spiked samples were between 92-107% for both proposed methods. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Simultaneous Detection of Sulfamethoxazole, Diclofenac, Carbamazepine, and Bezafibrate by Solid Phase Extraction and High Performance Liquid Chromatography with Diode Array Detection

    Science.gov (United States)

    Zhou, Z.; Jiang, J.-Q.

    2014-05-01

    A method of solid phase extraction (SPE) coupled with high performance liquid chromatography and diode array detection (HPLC-DAD) was studied for the simultaneous determination of sulfamethoxazole (SMX), diclofenac (DCF), carbamazepine (CBZ), and bezafi brate (BZF) in test solutions. The target compounds were extracted by SPE from samples, and the resulting elutes were analyzed using a HPLC-DAD system at wavelengths of 270, 280, 290, and 230 nm for SMX, DCF, CBZ, and BZF, respectively. This method shows good recoveries for SMX, DCF, CBZ, and BZF with mean recoveries of 89.7 ± 9.3%, 86.1 ± 7.6%, 95.0 ± 6.5%, and 94.0 ± 5.4%, respectively.

  13. Simultaneous Photoacoustic and Photopyroelectric Detection of Trace Gas Emissions from Some Plant Parts and Their Related Essential Oils in a Combined Detection Cell

    Science.gov (United States)

    Abu-Taha, M. I.; Abu-Teir, M. M.; Al-Jamal, A. J.; Eideh, H.

    The aim of this work was to establish the feasibility of the combined photoacoustic (PA) and photopyroelectric (PPE) detection of the vapours emitted from essential oils and their corresponding uncrushed leaves or flowers. Gas traces of jasmine (Jessamine (Jasminum)), mint (Mentha arvensis L.) and Damask rose (Rosa damascena Miller) and their essential oils were tested using a combined cell fitted with both a photopyroelectric film (PVDF) and a microphone in conjunction with a pulsed wideband infrared source (PWBS) source. Infrared PA and PPE absorbances were obtained simultaneously at room temperatures with excellent reproducibility and high signal-to-noise ratios. Significant similarities found between the PA and PPE spectra of the trace gas emissions of plant parts, i.e., flowers or leaves and their related essential oils show the good correlation of their emissions and that both effects are initiated by the same absorbing molecules.

  14. [Simultaneous separation and detection of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate by RP-HPLC and structure confirmation].

    Science.gov (United States)

    Zhao, Yan-Yan; Liu, Li-Yan; Han, Yuan-Yuan; Li, Yue-Qiu; Wang, Yan; Shi, Min-Jian

    2013-08-01

    A simple, fast and sensitive analytical method for the simultaneous separation and detection of 18alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B by RP-HPLC and drug quality standard was established. The structures of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate have been confirmed. Reference European Pharmacopoeia EP7.0 version, British Pharmacopoeia 2012 version, National Drug Standards of China (WS 1-XG-2002), domestic and international interrelated literature were referred to select the composition of mobile phase. The experimental parameters including salt concentration, pH, addition quantities of organic solvent, column temperature and flow rate were optimized. Finally, the assay was conducted on a Durashell-C18 column (250 mm x 4.6 mm, 5 microm) with 0.01 mol x mL(-1) ammonium perchlorate (add ammonia to adjust the pH value to 8.2) -methanol (48 : 52) as mobile phase at the flow rate of 0.8 mL x min(-1), and the detection wavelength was set at 254 nm. The column temperature was 50 degrees C and the injection volume was 10 microL. The MS, NMR, UV and RP-HPLC were used to confirm the structures of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate. Under the optimized separation conditions, the calibration curves of 18 alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B showed good linearity within the concentration of 0.50-100 microg x mL(-1) (r = 0.999 9). The detection limits for 18alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B were 0.15, 0.10, 0.10, 0.15 microg x mL(-1) respectively. The method is sensitive, reproducible and the results are accurate and reliable. It can be used for chiral resolution of 18alpha-glycyrrhizinic acid, 18Pbeta-glycyrrhizinic acid, and detection content of principal component and

  15. Detecting concealed information from groups using a dynamic questioning approach: simultaneous skin conductance measurement and immediate feedback

    Directory of Open Access Journals (Sweden)

    Ewout H Meijer

    2013-02-01

    Full Text Available Lie detection procedures typically aim at determining the guilt or innocence of a single suspect. The Concealed Information Test (CIT, for example, has been shown to be highly successful in detecting the presence or absence of crime-related information in a suspect’s memory. Many of today’s security threats, however, do not come from individuals, but from organized groups such as criminal organizations or terrorist networks. In this study, we tested whether a plan of an upcoming mock terrorist attack could be extracted from a group of suspects using a dynamic questioning approach. One-hundred participants were tested in 20 groups of 5. Each group was asked to plan a mock terrorist attack based on a list of potential countries, cities and streets. Next, three questions referring to the country, city, and street were presented, each with 5 options. Skin conductance in all 5 members of the group was measured simultaneously during this presentation. The dynamic questioning approach entailed direct analysis of the data, and if the average skin conductance of the group to a certain option exceeded a threshold, this option was followed up. E.g., if the reaction to the option ‘Italy’ exceeded the threshold, this was followed up by presenting 5 cities in Italy. Results showed that in 19 of the 20 groups the country was correctly detected using this procedure. In 13 of these remaining 19 groups the city was correctly detected. In 7 of these 13, the street was also correctly detected. The question about the country resulted in no false positives (out of 20, the question about the city resulted in 2 false positives (out of 19, while the question about the streets resulted in 2 false positives (out of 13. Furthermore, the 2 false positives at the city level also yielded a false positive at the street level. Taken together these results indicate our dynamic questioning approach can help to unveil plans about a mock terrorist attack.

  16. Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR.

    Science.gov (United States)

    Kuamsab, Napaporn; Putaporntip, Chaturong; Pattanawong, Urassaya; Jongwutiwes, Somchai

    2012-06-10

    Gametocyte carriage is essential for malaria transmission and endemicity of disease; thereby it is a target for malaria control strategies. Malaria-infected individuals may harbour gametocytes below the microscopic detection threshold that can be detected by reverse transcription polymerase chain reaction (RT-PCR) targeting gametocyte-specific mRNA. To date, RT-PCR has mainly been applied to the diagnosis of Plasmodium falciparum gametocytes but very limited for that of Plasmodium vivax. A multiplex-nested RT-PCR targeting Pfs25 and Pvs25 mRNA specific to mature gametocytes of P. falciparum and P. vivax, respectively, was developed. The assay was evaluated using blood samples collected in rainy and dry seasons from febrile patients,in a malaria-endemic area in Thailand. Malaria diagnosis was performed by Giemsa-stained blood smears and 18S rRNA PCR. The multiplex-nested RT-PCR detected Pfs25 mRNA in 75 of 86 (87.2%) P. falciparum-infected individuals and Pvs25 mRNA in 82 of 90 (91.1%) P. vivax malaria patients diagnosed by 18S rRNA PCR. Gametocytes were detected in 38 (eight P. falciparum and 30 P. vivax) of 157 microscopy positive samples, implying that a large number of patients harbour sub-microscopic gametocytaemia. No seasonal differences in gametocyte carriage were observed for both malaria species diagnosed by multiplex-nested RT-PCR. With single-nested RT-PCR targeting Pfs25 or Pvs25 mRNA as standard, the multiplex-nested RT-PCR offered sensitivities of 97.4% and 98.9% and specificities of 100% and 98.8% for diagnosing mature gametocytes of P. falciparum and P. vivax, respectively. The minimum detection limit of the multiplex-nested PCR was 10 copies of templates. The multiplex-nested RT-PCR developed herein is useful for simultaneous assessment of both P. falciparum and P. vivax gametocyte carriage that is prevalent and generally sympatric in several malaria-endemic areas outside Africa.

  17. One-step Multiplex RT-PCR Method for Simultaneous Detection of Seed Transmissible Bacteria and Viruses in Pepper and Tomato Seeds

    Directory of Open Access Journals (Sweden)

    Kyusik Jeong

    2011-04-01

    Full Text Available The aim of this study was to develop specific and sensitive PCR-based procedures for simultaneous detection of economically important plant seed infection pathogenic bacteria and virus, Xanthomonns campestris pv. vesicatoria (Xcv, Clavibacter michiganensis subsp. michiganensis (Cmm, Erwinia carotovora subsp. carotovora (Ecc, Pepper mild mottle virus (PMMoV and Tobacco mild green mosaic virus (TMGMV in pepper and tomato seeds. Most of pepper and tomato bacterial and virus diseases are responsible for germination and growth obstruction. PCR with arbitral primers: selection of specific primers, performance of PCR with specific primers and determination of the threshold level for pathogens detection. To detect simultaneously the Xcv, Cmm, Ecc, PMMoV and TMGMV in pepper and tomato seeds, five pairs (Cmm-F/R, Ecc-F/R, Xcv-F/R, PMMoV-F/R, TMGMV-F/R of specific primer were synthesized by primer-blast program. The multiplex PCR for the five pathogens in pepper and tomato seeds could detect specially without interference among primers and/or cDNA of plant seeds and other plant pathogens. The PCR result for pathogen detection using 20 commercial pepper and 10 tomato seed samples, Ecc was detected from 4 pepper and 2 tomato seed samples, PMMoV was detected from 1 pepper seed sample, and PMMoV and TMGMV were simultaneously detected from 1 pepper seed sample.

  18. One-step Multiplex RT-PCR Method for Simultaneous Detection of Seed Transmissible Bacterium and Virus Occurring on Brassicaceae Crop Seeds

    Directory of Open Access Journals (Sweden)

    Kyusik Jeong

    2011-04-01

    Full Text Available The aim of this research was to develop specific and sensitive PCR-based procedures for simultaneous detection of economically important plant pathogenic bacteria and seed borne virus in commercial Brassicaceae crop seeds, Xanthomonns campestris pv. campestris (Xcc and Lettuce Mosaic Virus (LMV. Bacterial and virus diseases of Brassicaceae leaves are responsible for heavy losses. PCR with arbitral primers: selection of specific primers, performance of PCR with specific primers and determination of the threshold level for pathogens detection. To detect simultaneously the Xcc and LMV in commercial Brassicaceae crop seeds (lettuce, kohlrabi, radish, chinese cabbage and cabbage, two pairs of specific primer (LMV-F/R, Xcc-F/R were synthesized by using primer-blast program (http://www.ncbi.nlm.nih.gov/tools/ primer-blast/. The multiplex PCR for the two pathogens in Brassicaceae crop seeds could detect specifically without interference among primers and/or cDNA of other plant pathogens. The pathogen detection limit was determined at 1 ng of RNA extracted from pathogens. In the total PCR results for pathogen detection using commercial kohlrabi (10 varieties, lettuce (50 varieties, radish (20 varieties, chinese cabbage (20 varieties and cabbage (20 varieties, LMV and Xcc were detected from 39 and 2 varieties, respectively. In the PCR result of lettuce, LMV and Xcc were simultaneously detected in 8 varieties.

  19. Single assay for simultaneous detection and differential identification of human and avian influenza virus types, subtypes, and emergent variants.

    Directory of Open Access Journals (Sweden)

    David Metzgar

    Full Text Available For more than four decades the cause of most type A influenza virus infections of humans has been attributed to only two viral subtypes, A/H1N1 or A/H3N2. In contrast, avian and other vertebrate species are a reservoir of type A influenza virus genome diversity, hosting strains representing at least 120 of 144 combinations of 16 viral hemagglutinin and 9 viral neuraminidase subtypes. Viral genome segment reassortments and mutations emerging within this reservoir may spawn new influenza virus strains as imminent epidemic or pandemic threats to human health and poultry production. Traditional methods to detect and differentiate influenza virus subtypes are either time-consuming and labor-intensive (culture-based or remarkably insensitive (antibody-based. Molecular diagnostic assays based upon reverse transcriptase-polymerase chain reaction (RT-PCR have short assay cycle time, and high analytical sensitivity and specificity. However, none of these diagnostic tests determine viral gene nucleotide sequences to distinguish strains and variants of a detected pathogen from one specimen to the next. Decision-quality, strain- and variant-specific pathogen gene sequence information may be critical for public health, infection control, surveillance, epidemiology, or medical/veterinary treatment planning. The Resequencing Pathogen Microarray (RPM-Flu is a robust, highly multiplexed and target gene sequencing-based alternative to both traditional culture- or biomarker-based diagnostic tests. RPM-Flu is a single, simultaneous differential diagnostic assay for all subtype combinations of type A influenza viruses and for 30 other viral and bacterial pathogens that may cause influenza-like illness. These other pathogen targets of RPM-Flu may co-infect and compound the morbidity and/or mortality of patients with influenza. The informative specificity of a single RPM-Flu test represents specimen-specific viral gene sequences as determinants of virus type, A

  20. Single assay for simultaneous detection and differential identification of human and avian influenza virus types, subtypes, and emergent variants.

    Science.gov (United States)

    Metzgar, David; Myers, Christopher A; Russell, Kevin L; Faix, Dennis; Blair, Patrick J; Brown, Jason; Vo, Scott; Swayne, David E; Thomas, Colleen; Stenger, David A; Lin, Baochuan; Malanoski, Anthony P; Wang, Zheng; Blaney, Kate M; Long, Nina C; Schnur, Joel M; Saad, Magdi D; Borsuk, Lisa A; Lichanska, Agnieszka M; Lorence, Matthew C; Weslowski, Brian; Schafer, Klaus O; Tibbetts, Clark

    2010-02-03

    For more than four decades the cause of most type A influenza virus infections of humans has been attributed to only two viral subtypes, A/H1N1 or A/H3N2. In contrast, avian and other vertebrate species are a reservoir of type A influenza virus genome diversity, hosting strains representing at least 120 of 144 combinations of 16 viral hemagglutinin and 9 viral neuraminidase subtypes. Viral genome segment reassortments and mutations emerging within this reservoir may spawn new influenza virus strains as imminent epidemic or pandemic threats to human health and poultry production. Traditional methods to detect and differentiate influenza virus subtypes are either time-consuming and labor-intensive (culture-based) or remarkably insensitive (antibody-based). Molecular diagnostic assays based upon reverse transcriptase-polymerase chain reaction (RT-PCR) have short assay cycle time, and high analytical sensitivity and specificity. However, none of these diagnostic tests determine viral gene nucleotide sequences to distinguish strains and variants of a detected pathogen from one specimen to the next. Decision-quality, strain- and variant-specific pathogen gene sequence information may be critical for public health, infection control, surveillance, epidemiology, or medical/veterinary treatment planning. The Resequencing Pathogen Microarray (RPM-Flu) is a robust, highly multiplexed and target gene sequencing-based alternative to both traditional culture- or biomarker-based diagnostic tests. RPM-Flu is a single, simultaneous differential diagnostic assay for all subtype combinations of type A influenza viruses and for 30 other viral and bacterial pathogens that may cause influenza-like illness. These other pathogen targets of RPM-Flu may co-infect and compound the morbidity and/or mortality of patients with influenza. The informative specificity of a single RPM-Flu test represents specimen-specific viral gene sequences as determinants of virus type, A/HN subtype, virulence

  1. Simultaneous detection of Fusarium culmorum and F. graminearum in plant material by duplex PCR with melting curve analysis.

    Science.gov (United States)

    Brandfass, Christoph; Karlovsky, Petr

    2006-01-23

    Fusarium head blight (FHB) is a disease of cereal crops, which has a severe impact on wheat and barley production worldwide. Apart from reducing the yield and impairing grain quality, FHB leads to contamination of grain with toxic secondary metabolites (mycotoxins), which pose a health risk to humans and livestock. The Fusarium species primarily involved in FHB are F. graminearum and F. culmorum. A key prerequisite for a reduction in the incidence of FHB is an understanding of its epidemiology. We describe a duplex-PCR-based method for the simultaneous detection of F. culmorum and F. graminearum in plant material. Species-specific PCR products are identified by melting curve analysis performed in a real-time thermocycler in the presence of the fluorescent dye SYBR Green I. In contrast to multiplex real-time PCR assays, the method does not use doubly labeled hybridization probes. PCR with product differentiation by melting curve analysis offers a cost-effective means of qualitative analysis for the presence of F. culmorum and F. graminearum in plant material. This method is particularly suitable for epidemiological studies involving a large number of samples.

  2. Monte Carlo simulation of simultaneous radiation detection in the hybrid tomography system ClearPET-XPAD3/CT

    Energy Technology Data Exchange (ETDEWEB)

    Dávila, H. Olaya, E-mail: hernan.olaya@uptc.edu.co; Martínez, S. A. [Physics Department, Universidad Pedagógica y Tecnológica de Colombia, Tunja-Colombia (Colombia); Sevilla, A. C., E-mail: acsevillam@unal.edu.co; Castro, H. F. [Physics Department, Universidad Nacional de Colombia, Bogotá D.C - Colombia (Colombia)

    2016-07-07

    Using the Geant4 based simulation framework SciFW1, a detailed simulation was performed for a detector array in the hybrid tomography prototype for small animals called ClearPET / XPAD, which was built in the Centre de Physique des Particules de Marseille. The detector system consists of an array of phoswich scintillation detectors: LSO (Lutetium Oxy-ortosilicate doped with cerium Lu{sub 2}SiO{sub 5}:Ce) and LuYAP (Lutetium Ortoaluminate of Yttrium doped with cerium Lu{sub 0.7}Y{sub 0.3}AlO{sub 3}:Ce) for Positron Emission Tomography (PET) and hybrid pixel detector XPAD for Computed Tomography (CT). Simultaneous acquisition of deposited energy and the corresponding time - position for each recorded event were analyzed, independently, for both detectors. interference between detection modules for PET and CT. Information about amount of radiation reaching each phoswich crystal and XPAD detector using a phantom in order to study the effectiveness by radiation attenuation and influence the positioning of the radioactive source {sup 22}Na was obtained. The simulation proposed will improve distribution of detectors rings and interference values will be taken into account in the new versions of detectors.

  3. Fast simultaneous determination of trimethoprim and sulfamethoxazole by capillary zone electrophoresis with capacitively coupled contactless conductivity detection.

    Science.gov (United States)

    da Silva, Iranaldo Santos; Vidal, Denis Tadeu Rajh; do Lago, Claudimir Lucio; Angnes, Lúcio

    2013-04-01

    The association of trimethoprim and sulfamethoxazole is a very effective with antibiotic properties, and commonly used in the treatment of a variety of infections. Due to the importance in diseases treatment of humans and also of animals, the development of methods for their quantification in commercial formulations is highly desirable. In the present study, a rapid method for simultaneous determination of these compounds using CE with capacitively coupled contactless conductivity detection was developed. A favorable working region for both analytes was from 12.5 to 200 μmol/L (linear responses with R > 0.999 for N = 5). Other parameters calculated were sensitivity (1.28 ± 0.10/1.45 ± 0.11) min/(μmol L), RSD (4.5%/2.0%), and LOD (1.1/3.3) μmol/L for trimethoprim and sulfamethoxazole, respectively. Under this condition, the total run time was only 2.6 min. The proposed method was applied to the determination of trimethoprim and sulfamethoxazole in commercial samples and the results were compared to those obtained by using a HPLC pharmacopoeia method. This new method is advantageous for quality-control analyses of trimethoprim and sulfamethoxazole in pharmaceuticals samples, because it is rapid and precise. Moreover, it is less laborious and demands minimum amounts of reagents in comparison to the recommended method. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Monte Carlo simulation of simultaneous radiation detection in the hybrid tomography system ClearPET-XPAD3/CT

    Science.gov (United States)

    Dávila, H. Olaya; Sevilla, A. C.; Castro, H. F.; Martínez, S. A.

    2016-07-01

    Using the Geant4 based simulation framework SciFW1, a detailed simulation was performed for a detector array in the hybrid tomography prototype for small animals called ClearPET / XPAD, which was built in the Centre de Physique des Particules de Marseille. The detector system consists of an array of phoswich scintillation detectors: LSO (Lutetium Oxy-ortosilicate doped with cerium Lu2SiO5:Ce) and LuYAP (Lutetium Ortoaluminate of Yttrium doped with cerium Lu0.7Y0.3AlO3:Ce) for Positron Emission Tomography (PET) and hybrid pixel detector XPAD for Computed Tomography (CT). Simultaneous acquisition of deposited energy and the corresponding time - position for each recorded event were analyzed, independently, for both detectors. interference between detection modules for PET and CT. Information about amount of radiation reaching each phoswich crystal and XPAD detector using a phantom in order to study the effectiveness by radiation attenuation and influence the positioning of the radioactive source 22Na was obtained. The simulation proposed will improve distribution of detectors rings and interference values will be taken into account in the new versions of detectors.

  5. Simultaneous detection of Zika, Chikungunya and Dengue viruses by a multiplex real-time RT-PCR assay.

    Science.gov (United States)

    Pabbaraju, Kanti; Wong, Sallene; Gill, Kara; Fonseca, Kevin; Tipples, Graham A; Tellier, Raymond

    2016-10-01

    In the recent past, arboviruses such as Chikungunya (CHIKV) and Zika (ZIKV) have increased their area of endemicity and presented as an emerging global public health threat. To design an assay for the simultaneous detection of ZIKV, CHIKV and Dengue (DENV) 1-4 from patients with symptoms of arboviral infection. This would be advantageous because of the similar clinical presentation typically encountered with these viruses and their co-circulation in endemic areas. In this study we have developed and validated a triplex real time reverse transcription PCR assay using hydrolysis probes targeting the non-structural 5 (NS5) region of ZIKV, non-structural protein 4 (nsP4) from CHIKV and 3' untranslated region (3'UTR) of DENV 1-4. The 95% LOD by the triplex assay was 15 copies/reaction for DENV-1 and less than 10 copies/reaction for all other viruses. The triplex assay was 100% specific and did not amplify any of the other viruses tested. The assay was reproducible and adaptable to testing different specimen types including serum, plasma, urine, placental tissue, brain tissue and amniotic fluid. This assay can be easily implemented for diagnostic testing of patient samples, even in a high throughput laboratory. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. [Simultaneous determination of five effective components in Sijunzi bolus using high performance liquid chromatography-evaporation light scattering detection].

    Science.gov (United States)

    Li, Chunying; Zhang, Xiaojun

    2010-01-01

    A high performance liquid chromatographic (HPLC) method was developed for the simultaneous determination of lobetyolin, pachymic acid, glycyrrhizic acid, atractylenoide III and atractylenolide I in Sijunzi bolus. The separation was performed on an HIQ SIL C18 V column (250 mm x 4.6 mm, 5 microm) with 0.5% acetic acid-methanol as the mobile phase of gradient elution at a flow rate of 1.0 mL/min. The detection was performed with an evaporation light scattering detector (ELSD) and the sample volume was 10 microL. The temperature of drift tube and heating grade of nebulizer was respectively set at 55 degrees C and 60% at 0.2 MPa of pressure. Nitrogen gas was used as carrier gas. Under the optimized conditions, there were good linear relationships between the logarithm values of mass concentration and the peak areas of lobetyolin, pachymic acid, glycyrrhizic acid, atractylenoide III and atractylenolide I in the ranges of 0.076 - 1.21, 0.048 -0.76, 0.153 - 2.45, 0.045 - 0.72 and 0.098 - 1.56 g/L, respectively. The recoveries of the five components were between 97.13% and 100.25%, the relative standard deviations (RSDs) were between 1.23% and 2.44%. This method is simple, rapid, accurate and suitable for the quality control of Sijunzi bolus.

  7. Simultaneous detection of Fusarium culmorum and F. graminearum in plant material by duplex PCR with melting curve analysis

    Directory of Open Access Journals (Sweden)

    Karlovsky Petr

    2006-01-01

    Full Text Available Abstract Background Fusarium head blight (FHB is a disease of cereal crops, which has a severe impact on wheat and barley production worldwide. Apart from reducing the yield and impairing grain quality, FHB leads to contamination of grain with toxic secondary metabolites (mycotoxins, which pose a health risk to humans and livestock. The Fusarium species primarily involved in FHB are F. graminearum and F. culmorum. A key prerequisite for a reduction in the incidence of FHB is an understanding of its epidemiology. Results We describe a duplex-PCR-based method for the simultaneous detection of F. culmorum and F. graminearum in plant material. Species-specific PCR products are identified by melting curve analysis performed in a real-time thermocycler in the presence of the fluorescent dye SYBR Green I. In contrast to multiplex real-time PCR assays, the method does not use doubly labeled hybridization probes. Conclusion PCR with product differentiation by melting curve analysis offers a cost-effective means of qualitative analysis for the presence of F. culmorum and F. graminearum in plant material. This method is particularly suitable for epidemiological studies involving a large number of samples.

  8. A new method for simultaneous detection and discrimination of Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) using real time PCR with high resolution melting (HRM) analysis.

    Science.gov (United States)

    Marin, M S; Quintana, S; Leunda, M R; Recavarren, M; Pagnuco, I; Späth, E; Pérez, S; Odeón, A

    2016-01-01

    Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are antigenically and genetically similar. The aim of this study was to develop a simple and reliable one-step real time PCR assay with high resolution melting (HRM) analysis for the simultaneous detection and differentiation of BoHV-1 and BoHV-5. Optimization of assay conditions was performed with DNA from reference strains. Then, DNA from field isolates, clinical samples and tissue samples of experimentally infected animals were studied by real time PCR-HRM. An efficient amplification of real time PCR products was obtained, and a clear melting curve and appropriate melting peaks for both viruses were achieved in the HRM curve analysis for BoHV type identification. BoHV was identified in all of the isolates and clinical samples, and BoHV types were properly differentiated. Furthermore, viral DNA was detected in 12/18 and 7/18 samples from BoHV-1- and BoHV-5-infected calves, respectively. Real time PCR-HRM achieved a higher sensitivity compared with virus isolation or conventional PCR. In this study, HRM was used as a novel procedure. This method provides rapid, sensitive, specific and simultaneous detection of bovine alpha-herpesviruses DNA. Thus, this technique is an excellent tool for diagnosis, research and epidemiological studies of these viruses in cattle. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Reliability of ultrasonography in detecting shoulder disease in patients with rheumatoid arthritis.

    LENUS (Irish Health Repository)

    Bruyn, G A W

    2009-03-01

    To assess the intra and interobserver reproducibility of musculoskeletal ultrasonography (US) among rheumatologists in detecting destructive and inflammatory shoulder abnormalities in patients with rheumatoid arthritis (RA) and to determine the overall agreement between US and MRI.

  10. Reliable fault detection and diagnosis of photovoltaic systems based on statistical monitoring approaches

    KAUST Repository

    Harrou, Fouzi; Sun, Ying; Taghezouit, Bilal; Saidi, Ahmed; Hamlati, Mohamed-Elkarim

    2017-01-01

    This study reports the development of an innovative fault detection and diagnosis scheme to monitor the direct current (DC) side of photovoltaic (PV) systems. Towards this end, we propose a statistical approach that exploits the advantages of one

  11. A simple and reliable methodology to detect egg white in art samples

    Indian Academy of Sciences (India)

    2013-04-26

    Apr 26, 2013 ... threshold density values useful for the detection of ovalbumin in samples from ancient works of art. .... slides a mixture of a water solution of dry egg white and the .... ily, facing the problems of sample leakage, background.

  12. Reliability of ultrasonography in detecting shoulder disease in patients with rheumatoid arthritis

    NARCIS (Netherlands)

    Bruyn, G. A. W.; Naredo, E.; Moeller, I.; Moragues, C.; Garrido, J.; de Bock, G. H.; d'Agostino, M-A; Filippucci, E.; Iagnocco, A.; Backhaus, M.; Swen, W. A. A.; Balint, P.; Pineda, C.; Milutinovic, S.; Kane, D.; Kaeley, G.; Narvaez, F. J.; Wakefield, R. J.; Narvaez, J. A.; de Augustin, J.; Schmidt, W. A.; Moller, I.; Swen, N.; de Agustin, J.

    Objective: To assess the intra and interobserver reproducibility of musculoskeletal ultrasonography ( US) among rheumatologists in detecting destructive and inflammatory shoulder abnormalities in patients with rheumatoid arthritis ( RA) and to determine the overall agreement between US and MRI.

  13. Novel Multiplex PCR Assay for Detection of Chlorhexidine-Quaternary Ammonium, Mupirocin, and Methicillin Resistance Genes, with Simultaneous Discrimination of Staphylococcus aureus from Coagulase-Negative Staphylococci.

    Science.gov (United States)

    McClure, Jo-Ann; Zaal DeLongchamp, Johanna; Conly, John M; Zhang, Kunyan

    2017-06-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a clinically significant pathogen that is resistant to a wide variety of antibiotics and responsible for a large number of nosocomial infections worldwide. The Agency for Healthcare Research and Quality and the Centers for Disease Control and Prevention recently recommended the adoption of universal mupirocin-chlorhexidine decolonization of all admitted intensive care unit patients rather than MRSA screening with targeted treatments, which raises a serious concern about the selection of resistance to mupirocin and chlorhexidine in strains of staphylococci. Thus, a simple, rapid, and reliable approach is paramount in monitoring the prevalence of resistance to these agents. We developed a simple multiplex PCR assay capable of screening Staphylococcus isolates for the presence of antiseptic resistance genes for chlorhexidine and quaternary ammonium compounds, as well as mupirocin and methicillin resistance genes, while simultaneously discriminating S. aureus from coagulase-negative staphylococci (CoNS). The assay incorporates 7 PCR targets, including the Staphylococcus 16S rRNA gene (specifically detecting Staphylococcus spp.), nuc (distinguishing S. aureus from CoNS), mecA (distinguishing MRSA from methicillin-susceptible S. aureus ), mupA and mupB (identifying high-level mupirocin resistance), and qac and smr (identifying chlorhexidine and quaternary ammonium resistance). Our assay demonstrated 100% sensitivity, specificity, and accuracy in a total of 23 variant antiseptic- and/or antibiotic-resistant control strains. Further validation of our assay using 378 randomly selected and previously well-characterized local clinical isolates confirmed its feasibility and practicality. This may prove to be a useful tool for multidrug-resistant Staphylococcus monitoring in clinical laboratories, particularly in the wake of increased chlorhexidine and mupirocin treatments. Copyright © 2017 American Society for Microbiology.

  14. Simultaneous detection of six urinary pteridines and creatinine by high-performance liquid chromatography-tandem mass spectrometry for clinical breast cancer detection.

    Science.gov (United States)

    Burton, Casey; Shi, Honglan; Ma, Yinfa

    2013-11-19

    Recent preliminary studies have implicated urinary pteridines as candidate biomarkers in a growing number of malignancies including breast cancer. While the developments of capillary electrophoresis-laser induced fluorescence (CE-LIF), high performance liquid chromatography (HPLC), and liquid chromatography-mass spectroscopy (LC-MS) pteridine urinalyses among others have helped to enable these findings, limitations including poor pteridine specificity, asynchronous or nonexistent renal dilution normalization, and a lack of information regarding adduct formation in mass spectrometry techniques utilizing electrospray ionization (ESI) have prevented application of these techniques to a larger clinical setting. In this study, a simple, rapid, specific, and sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method has been developed and optimized for simultaneous detection of six pteridines previously implicated in breast cancer and creatinine as a renal dilution factor in urine. In addition, this study reports cationic adduct formation of urinary pteridines under ESI-positive ionization for the first time. This newly developed technique separates and detects the following six urinary pteridines: 6-biopterin, 6-hydroxymethylpterin, d-neopterin, pterin, isoxanthopterin, and xanthopterin, as well as creatinine. The method detection limit for the pteridines is between 0.025 and 0.5 μg/L, and for creatinine, it is 0.15 μg/L. The method was also validated by spiked recoveries (81-105%), reproducibility (RSD: 1-6%), and application to 25 real urine samples from breast cancer positive and negative samples through a double-blind study. The proposed technique was finally compared directly with a previously reported CE-LIF technique, concluding that additional or alternative renal dilution factors are needed for proper investigation of urinary pteridines as breast cancer biomarkers.

  15. Implanted cardiac devices are reliably detected by commercially available metal detectors

    DEFF Research Database (Denmark)

    Holm, Katja Fiedler; Hjortshøj, Søren Pihlkjær; Pehrson, Steen

    2013-01-01

    Explosions of Cardiovascular Implantable Electronic Devices (CIEDs) (pacemakers, defibrillators, and loop recorders) are a well-recognized problem during cremation, due to lithium-iodine batteries. In addition, burial of the deceased with a CIED can present a potential risk for environmental...... contamination. Therefore, detection of CIEDs in the deceased would be of value. This study evaluated a commercially available metal detector for detecting CIEDs....

  16. Experimental Research of Reliability of Plant Stress State Detection by Laser-Induced Fluorescence Method

    Directory of Open Access Journals (Sweden)

    Yury Fedotov

    2016-01-01

    Full Text Available Experimental laboratory investigations of the laser-induced fluorescence spectra of watercress and lawn grass were conducted. The fluorescence spectra were excited by YAG:Nd laser emitting at 532 nm. It was established that the influence of stress caused by mechanical damage, overwatering, and soil pollution is manifested in changes of the spectra shapes. The mean values and confidence intervals for the ratio of two fluorescence maxima near 685 and 740 nm were estimated. It is presented that the fluorescence ratio could be considered a reliable characteristic of plant stress state.

  17. Autism detection in early childhood (ADEC): reliability and validity data for a Level 2 screening tool for autistic disorder.

    Science.gov (United States)

    Nah, Yong-Hwee; Young, Robyn L; Brewer, Neil; Berlingeri, Genna

    2014-03-01

    The Autism Detection in Early Childhood (ADEC; Young, 2007) was developed as a Level 2 clinician-administered autistic disorder (AD) screening tool that was time-efficient, suitable for children under 3 years, easy to administer, and suitable for persons with minimal training and experience with AD. A best estimate clinical Diagnostic and Statistical Manual of Mental Disorders (4th ed., text rev.; DSM-IV-TR; American Psychiatric Association, 2000) diagnosis of AD was made for 70 children using all available information and assessment results, except for the ADEC data. A screening study compared these children on the ADEC with 57 children with other developmental disorders and 64 typically developing children. Results indicated high internal consistency (α = .91). Interrater reliability and test-retest reliability of the ADEC were also adequate. ADEC scores reliably discriminated different diagnostic groups after controlling for nonverbal IQ and Vineland Adaptive Behavior Composite scores. Construct validity (using exploratory factor analysis) and concurrent validity using performance on the Autism Diagnostic Observation Schedule (Lord et al., 2000), the Autism Diagnostic Interview-Revised (Le Couteur, Lord, & Rutter, 2003), and DSM-IV-TR criteria were also demonstrated. Signal detection analysis identified the optimal ADEC cutoff score, with the ADEC identifying all children who had an AD (N = 70, sensitivity = 1.0) but overincluding children with other disabilities (N = 13, specificity ranging from .74 to .90). Together, the reliability and validity data indicate that the ADEC has potential to be established as a suitable and efficient screening tool for infants with AD. 2014 APA

  18. Technical Note: The single particle soot photometer fails to reliably detect PALAS soot nanoparticles

    Directory of Open Access Journals (Sweden)

    M. Gysel

    2012-12-01

    Full Text Available The single particle soot photometer (SP2 uses laser-induced incandescence (LII for the measurement of atmospheric black carbon (BC particles. The BC mass concentration is obtained by combining quantitative detection of BC mass in single particles with a counting efficiency of 100% above its lower detection limit. It is commonly accepted that a particle must contain at least several tenths of a femtogram BC in order to be detected by the SP2.

    Here we show the result that most BC particles from a PALAS spark discharge soot generator remain undetected by the SP2, even if their BC mass, as independently determined with an aerosol particle mass analyser (APM, is clearly above the typical lower detection limit of the SP2. Comparison of counting efficiency and effective density data of PALAS soot with flame generated soot (combustion aerosol standard burner, CAST, fullerene soot and carbon black particles (Cabot Regal 400R reveals that particle morphology can affect the SP2's lower detection limit. PALAS soot particles are fractal-like agglomerates of very small primary particles with a low fractal dimension, resulting in a very low effective density. Such loosely packed particles behave like "the sum of individual primary particles" in the SP2's laser. Accordingly, most PALAS soot particles remain undetected as the SP2's laser intensity is insufficient to heat the primary particles to their vaporisation temperature because of their small size (Dpp ≈ 5–10 nm. Previous knowledge from pulsed laser-induced incandescence indicated that particle morphology might have an effect on the SP2's lower detection limit, however, an increase of the lower detection limit by a factor of ∼5–10, as reported here for PALAS soot, was not expected.

    In conclusion, the SP2's lower detection limit at a certain laser power depends primarily on the total BC mass per particle for compact particles with sufficiently high effective

  19. Test-retest reliability and minimal detectable change of two simplified 3-point balance measures in patients with stroke.

    Science.gov (United States)

    Chen, Yi-Miau; Huang, Yi-Jing; Huang, Chien-Yu; Lin, Gong-Hong; Liaw, Lih-Jiun; Lee, Shih-Chieh; Hsieh, Ching-Lin

    2017-10-01

    The 3-point Berg Balance Scale (BBS-3P) and 3-point Postural Assessment Scale for Stroke Patients (PASS-3P) were simplified from the BBS and PASS to overcome the complex scoring systems. The BBS-3P and PASS-3P were more feasible in busy clinical practice and showed similarly sound validity and responsiveness to the original measures. However, the reliability of the BBS-3P and PASS-3P is unknown limiting their utility and the interpretability of scores. We aimed to examine the test-retest reliability and minimal detectable change (MDC) of the BBS-3P and PASS-3P in patients with stroke. Cross-sectional study. The rehabilitation departments of a medical center and a community hospital. A total of 51 chronic stroke patients (64.7% male). Both balance measures were administered twice 7 days apart. The test-retest reliability of both the BBS-3P and PASS-3P were examined by intraclass correlation coefficients (ICC). The MDC and its percentage over the total score (MDC%) of each measure was calculated for examining the random measurement errors. The ICC values of the BBS-3P and PASS-3P were 0.99 and 0.97, respectively. The MDC% (MDC) of the BBS-3P and PASS-3P were 9.1% (5.1 points) and 8.4% (3.0 points), respectively, indicating that both measures had small and acceptable random measurement errors. Our results showed that both the BBS-3P and the PASS-3P had good test-retest reliability, with small and acceptable random measurement error. These two simplified 3-level balance measures can provide reliable results over time. Our findings support the repeated administration of the BBS-3P and PASS-3P to monitor the balance of patients with stroke. The MDC values can help clinicians and researchers interpret the change scores more precisely.

  20. Reliability of tensiomyography and myotonometry in detecting mechanical and contractile characteristics of the lumbar erector spinae in healthy volunteers.

    Science.gov (United States)

    Lohr, Christine; Braumann, Klaus-Michael; Reer, Ruediger; Schroeder, Jan; Schmidt, Tobias

    2018-04-20

    Tensiomyography™ (TMG) and MyotonPRO ® (MMT) are two non-invasive devices for monitoring muscle contractile and mechanical characteristics. This study aimed to evaluate the test-retest reliability of TMG and MMT parameters for measuring (TMG:) muscle displacement (D m ), contraction time (T c ), and velocity (V c ) and (MMT:) frequency (F), stiffness (S), and decrement (D) of the erector spinae muscles (ES) in healthy adults. A particular focus was set on the establishment of reliability measures for the previously barely evaluated secondary TMG parameter V c . Twenty-four subjects (13 female and 11 male, mean ± SD, 38.0 ± 12.0 years) were measured using TMG and MMT over 2 consecutive days. Absolute and relative reliability was calculated by standard error of measurement (SEM, SEM%), Minimum detectable change (MDC, MDC%), coefficient of variation (CV%) and intraclass correlation coefficient (ICC, 3.1) with a 95% confidence interval (CI). The ICCs for all variables and test-retest intervals ranged from 0.75 to 0.99 indicating a good to excellent relative reliability for both TMG and MMT, demonstrating the lowest values for TMG T c and between-day MMT D (ICC TMG parameter (ICC > 0.95, CV TMG V c could be established successfully. Its further applicability needs to be confirmed in future studies. MMT was found to be more reliable on repeated testing than the two other TMG parameters D m and T c .

  1. Three dimensional quantitative coronary angiography can detect reliably ischemic coronary lesions based on fractional flow reserve.

    Science.gov (United States)

    Chung, Woo-Young; Choi, Byoung-Joo; Lim, Seong-Hoon; Matsuo, Yoshiki; Lennon, Ryan J; Gulati, Rajiv; Sandhu, Gurpreet S; Holmes, David R; Rihal, Charanjit S; Lerman, Amir

    2015-06-01

    Conventional coronary angiography (CAG) has limitations in evaluating lesions producing ischemia. Three dimensional quantitative coronary angiography (3D-QCA) shows reconstructed images of CAG using computer based algorithm, the Cardio-op B system (Paieon Medical, Rosh Ha'ayin, Israel). The aim of this study was to evaluate whether 3D-QCA can reliably predict ischemia assessed by myocardial fractional flow reserve (FFR) < 0.80. 3D-QCA images were reconstructed from CAG which also were evaluated with FFR to assess ischemia. Minimal luminal diameter (MLD), percent diameter stenosis (%DS), minimal luminal area (MLA), and percent area stenosis (%AS) were obtained. The results of 3D-QCA and FFR were compared. A total of 266 patients was enrolled for the present study. FFR for all lesions ranged from 0.57 to 1.00 (0.85 ± 0.09). Measurement of MLD, %DS, MLA, and %AS all were significantly correlated with FFR (r = 0.569, 0609, 0.569, 0.670, respectively, all P < 0.001). In lesions with MLA < 4.0 mm(2), %AS of more than 65.5% had a 80% sensitivity and a 83% specificity to predict FFR < 0.80 (area under curve, AUC was 0.878). 3D-QCA can reliably predict coronary lesions producing ischemia and may be used to guide therapeutic approach for coronary artery disease.

  2. Reliable fault detection and diagnosis of photovoltaic systems based on statistical monitoring approaches

    KAUST Repository

    Harrou, Fouzi

    2017-09-18

    This study reports the development of an innovative fault detection and diagnosis scheme to monitor the direct current (DC) side of photovoltaic (PV) systems. Towards this end, we propose a statistical approach that exploits the advantages of one-diode model and those of the univariate and multivariate exponentially weighted moving average (EWMA) charts to better detect faults. Specifically, we generate array\\'s residuals of current, voltage and power using measured temperature and irradiance. These residuals capture the difference between the measurements and the predictions MPP for the current, voltage and power from the one-diode model, and use them as fault indicators. Then, we apply the multivariate EWMA (MEWMA) monitoring chart to the residuals to detect faults. However, a MEWMA scheme cannot identify the type of fault. Once a fault is detected in MEWMA chart, the univariate EWMA chart based on current and voltage indicators is used to identify the type of fault (e.g., short-circuit, open-circuit and shading faults). We applied this strategy to real data from the grid-connected PV system installed at the Renewable Energy Development Center, Algeria. Results show the capacity of the proposed strategy to monitors the DC side of PV systems and detects partial shading.

  3. Fast Metabolite Identification in Nuclear Magnetic Resonance Metabolomic Studies: Statistical Peak Sorting and Peak Overlap Detection for More Reliable Database Queries.

    Science.gov (United States)

    Hoijemberg, Pablo A; Pelczer, István

    2018-01-05

    A lot of time is spent by researchers in the identification of metabolites in NMR-based metabolomic studies. The usual metabolite identification starts employing public or commercial databases to match chemical shifts thought to belong to a given compound. Statistical total correlation spectroscopy (STOCSY), in use for more than a decade, speeds the process by finding statistical correlations among peaks, being able to create a better peak list as input for the database query. However, the (normally not automated) analysis becomes challenging due to the intrinsic issue of peak overlap, where correlations of more than one compound appear in the STOCSY trace. Here we present a fully automated methodology that analyzes all STOCSY traces at once (every peak is chosen as driver peak) and overcomes the peak overlap obstacle. Peak overlap detection by clustering analysis and sorting of traces (POD-CAST) first creates an overlap matrix from the STOCSY traces, then clusters the overlap traces based on their similarity and finally calculates a cumulative overlap index (COI) to account for both strong and intermediate correlations. This information is gathered in one plot to help the user identify the groups of peaks that would belong to a single molecule and perform a more reliable database query. The simultaneous examination of all traces reduces the time of analysis, compared to viewing STOCSY traces by pairs or small groups, and condenses the redundant information in the 2D STOCSY matrix into bands containing similar traces. The COI helps in the detection of overlapping peaks, which can be added to the peak list from another cross-correlated band. POD-CAST overcomes the generally overlooked and underestimated presence of overlapping peaks and it detects them to include them in the search of all compounds contributing to the peak overlap, enabling the user to accelerate the metabolite identification process with more successful database queries and searching all tentative

  4. Clinical Application of a Multiplex Real-Time PCR Assay for Simultaneous Detection of Legionella Species, Legionella pneumophila, and Legionella pneumophila Serogroup 1

    OpenAIRE

    Benitez, Alvaro J.; Winchell, Jonas M.

    2014-01-01

    We developed a single-tube multiplex real-time PCR assay capable of simultaneously detecting and discriminating Legionella spp., Legionella pneumophila, and Legionella pneumophila serogroup 1 in primary specimens. Evaluation of 21 clinical specimens and 115 clinical isolates demonstrated this assay to be a rapid, high-throughput diagnostic test with 100% specificity that may aid during legionellosis outbreaks and epidemiologic investigations.

  5. Development of SCAR markers and PCR assays for single or simultaneous species-specific detection of Phytophthora nicotianae and Pythium helicoides in ebb-and-flow irrigated kalanchoe.

    Science.gov (United States)

    Ahonsi, Monday O; Ling, Yin; Kageyama, Koji

    2010-11-01

    Phytophthora nicotianae and Pythium helicoides are important water-borne oomycete pathogens of irrigated ornamentals particularly ebb-and-flow irrigated kalanchoe in Japan. We developed novel PCR-based sequence characterized amplified region markers and assays for rapid identification and species-specific detection of both pathogens in separate PCR reactions or simultaneously in a duplex PCR.

  6. Development of a sensitive real-time PCR for simultaneous detection and subtyping of influenza A and B viruses

    Directory of Open Access Journals (Sweden)

    Daniela Amicizia

    2005-03-01

    Full Text Available

    A new real-time PCR assay, using melting curve analysis, was developed for the rapid and reliable detection and sub-typing of influenza A and B.

    In order to evaluate it’s specificity, cell culture surnatants positive for Respiratory Syncytial Virus, Parainfluenza Viruses 1, 2 and 3, Measles Virus, Influenza A (to evaluate Influenza B primer and B (to evaluate Influenza A primer were tested and all of the results were negative.

    A series of Influenza A and B cell culture-grown viruses were diluted in virus transport medium, titrated and tested to determine the analytical sensibility which equated to 0.64, 0.026, 0.64, 0.62 PFU for A/H1N1, A/H3N2, Victoria-like and Yamagata-like B viruses, respectively. Twenty-five specimens, collected during the 2001/02 and 2002/03 seasons, which were positive for A/H1N1 (n = 7, A/H3N2 (n = 10, B Victoria-lineage (n = 5 and B Yamagata-lineage (n = 3, were tested in order to evaluate the assay’s clinical sensitivity, all of the results were positive.

    The new real-time PCR appears to be a suitable tool for virological surveillance and the diagnosis of respiratory infections.

  7. Reliability and validity of the KIPPPI: an early detection tool for psychosocial problems in toddlers.

    Directory of Open Access Journals (Sweden)

    Ingrid Kruizinga

    Full Text Available BACKGROUND: The KIPPPI (Brief Instrument Psychological and Pedagogical Problem Inventory is a Dutch questionnaire that measures psychosocial and pedagogical problems in 2-year olds and consists of a KIPPPI Total score, Wellbeing scale, Competence scale, and Autonomy scale. This study examined the reliability, validity, screening accuracy and clinical application of the KIPPPI. METHODS: Parents of 5959 2-year-old children in the Rotterdam area, the Netherlands, were invited to participate in the study. Parents of 3164 children (53.1% of all invited parents completed the questionnaire. The internal consistency was evaluated and in subsamples the test-retest reliability and concurrent validity with regard to the Child Behavioral Checklist (CBCL. Discriminative validity was evaluated by comparing scores of parents who worried about their child's upbringing and parent's that did not. Screening accuracy of the KIPPPI was evaluated against the CBCL by calculating the Receiver Operating Characteristic (ROC curves. The clinical application was evaluated by the relation between KIPPPI scores and the clinical decision made by the child health professionals. RESULTS: Psychometric properties of the KIPPPI Total score, Wellbeing scale, Competence scale and Autonomy scale were respectively: Cronbach's alphas: 0.88, 0.86, 0.83, 0.58. Test-retest correlations: 0.80, 0.76, 0.73, 0.60. Concurrent validity was as hypothesised. The KIPPPI was able to discriminate between parents that worried about their child and parents that did not. Screening accuracy was high (>0.90 for the KIPPPI Total score and for the Wellbeing scale. The KIPPPI scale scores and clinical decision of the child health professional were related (p<0.05, indicating a good clinical application. CONCLUSION: The results in this large-scale study of a diverse general population sample support the reliability, validity and clinical application of the KIPPPI Total score, Wellbeing scale and Competence

  8. Acoustic feedwater heater leak detection: Industry application of low ampersand high frequency detection increases response and reliability

    International Nuclear Information System (INIS)

    Woyshner, W.S.; Bryson, T.; Robertson, M.O.

    1993-01-01

    The Electric Power Research Institute has sponsored research associated with acoustic Feedwater Heater Leak Detection since the early 1980s. Results indicate that this technology is economically beneficial and dependable. Recent research work has employed acoustic sensors and signal conditioning with wider frequency range response and background noise elimination techniques to provide increased accuracy and dependability. Dual frequency sensors have been applied at a few facilities to provide information on this application of dual frequency response. Sensor mounting methods and attenuation due to various mounting configurations are more conclusively understood. These are depicted and discussed in detail. The significance of trending certain plant parameters such as heat cycle flows, heater vent and drain valve position, proper relief valve operation, etc. is also addressed. Test data were collected at various facilities to monitor the effect of varying several related operational parameters. A group of FWHLD Users have been involved from the inception of the project and reports on their latest successes and failures, along with various data depicting early detection of FWHLD tube leaks, will be included. 3 refs., 12 figs., 1 tab

  9. FISHing for bacteria in food--a promising tool for the reliable detection of pathogenic bacteria?

    Science.gov (United States)

    Rohde, Alexander; Hammerl, Jens Andre; Appel, Bernd; Dieckmann, Ralf; Al Dahouk, Sascha

    2015-04-01

    Foodborne pathogens cause millions of infections every year and are responsible for considerable economic losses worldwide. The current gold standard for the detection of bacterial pathogens in food is still the conventional cultivation following standardized and generally accepted protocols. However, these methods are time-consuming and do not provide fast information about food contaminations and thus are limited in their ability to protect consumers in time from potential microbial hazards. Fluorescence in situ hybridization (FISH) represents a rapid and highly specific technique for whole-cell detection. This review aims to summarize the current data on FISH-testing for the detection of pathogenic bacteria in different food matrices and to evaluate its suitability for the implementation in routine testing. In this context, the use of FISH in different matrices and their pretreatment will be presented, the sensitivity and specificity of FISH tests will be considered and the need for automation shall be discussed as well as the use of technological improvements to overcome current hurdles for a broad application in monitoring food safety. In addition, the overall economical feasibility will be assessed in a rough calculation of costs, and strengths and weaknesses of FISH are considered in comparison with traditional and well-established detection methods. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  10. Multivariate normative comparison, a novel method for more reliably detecting cognitive impairment in HIV infection

    NARCIS (Netherlands)

    Su, Tanja; Schouten, Judith; Geurtsen, Gert J.; Wit, Ferdinand W.; Stolte, Ineke G.; Prins, Maria; Portegies, Peter; Caan, Matthan W. A.; Reiss, Peter; Majoie, Charles B.; Schmand, Ben A.

    2015-01-01

    The objective of this study is to assess whether multivariate normative comparison (MNC) improves detection of HIV-1-associated neurocognitive disorder (HAND) as compared with Frascati and Gisslén criteria. One-hundred and three HIV-1-infected men with suppressed viremia on combination

  11. Reliability of using retinal vascular fractal dimension as a biomarker in the diabetic retinopathy detection

    NARCIS (Netherlands)

    Huang, F.; Dashtbozorg, B.; Zhang, J.; Bekkers, E.J.; Abbasi-Sureshjani, S.; Berendschot, T.T.J.M.; ter Haar Romenij, B.M.

    2016-01-01

    The retinal fractal dimension (FD) is a measure of vasculature branching pattern complexity. FD has been considered as a potential biomarker for the detection of several diseases like diabetes and hypertension. However, conflicting findings were found in the reported literature regarding the

  12. Comparison of specificity and sensitivity of immunochemical and molecular techniques for reliable detection of Erwinia amylovora

    Czech Academy of Sciences Publication Activity Database

    Kokošková, B.; Mráz, Ivan; Hýblová, Jana

    2007-01-01

    Roč. 52, č. 2 (2007), s. 175-182 ISSN 0015-5632 R&D Projects: GA AV ČR(CZ) 1QS500510558 Institutional research plan: CEZ:AV0Z50510513 Keywords : Erwinia amylovora * detection Subject RIV: EE - Microbiology, Virology Impact factor: 0.989, year: 2007

  13. Sensitive and reliable detection of genomic imbalances in human neuroblastomas using comparative genomic hybridisation analysis

    NARCIS (Netherlands)

    van Gele, M.; van Roy, N.; Jauch, A.; Laureys, G.; Benoit, Y.; Schelfhout, V.; de Potter, C. R.; Brock, P.; Uyttebroeck, A.; Sciot, R.; Schuuring, E.; Versteeg, R.; Speleman, F.

    1997-01-01

    Deletions of the short arm of chromosome 1, extra copies of chromosome 17q and MYCN amplification are the most frequently encountered genetic changes in neuroblastomas. Standard techniques for detection of one or more of these genetic changes are karyotyping, FISH analysis and LOH analysis by

  14. Indirect immunofluorescence assay for the simultaneous detection of antibodies against clinically important old and new world hantaviruses.

    Directory of Open Access Journals (Sweden)

    Sabine Lederer

    Full Text Available In order to detect serum antibodies against clinically important Old and New World hantaviruses simultaneously, multiparametric indirect immunofluorescence assays (IFAs based on biochip mosaics were developed. Each of the mosaic substrates consisted of cells infected with one of the virus types Hantaan (HTNV, Puumala (PUUV, Seoul (SEOV, Saaremaa (SAAV, Dobrava (DOBV, Sin Nombre (SNV or Andes (ANDV. For assay evaluation, serum IgG and IgM antibodies were analyzed using 184 laboratory-confirmed hantavirus-positive sera collected at six diagnostic centers from patients actively or previously infected with the following hantavirus serotypes: PUUV (Finland, n=97; SEOV (China, n=5; DOBV (Romania, n=7; SNV (Canada, n=23; ANDV (Argentina and Chile, n=52. The control panel comprised 89 sera from healthy blood donors. According to the reference tests, all 184 patient samples were seropositive for hantavirus-specific IgG (n=177; 96% and/or IgM (n=131; 72%, while all control samples were tested negative. In the multiparametric IFA applied in this study, 183 (99% of the patient sera were IgG and 131 (71% IgM positive (accordance with the reference tests: IgG, 96%; IgM, 93%. Overall IFA sensitivity for combined IgG and IgM analysis amounted to 100% for all serotypes, except for SNV (96%. Of the 89 control sera, 2 (2% showed IgG reactivity against the HTNV substrate, but not against any other hantavirus. Due to the high cross-reactivity of hantaviral nucleocapsid proteins, endpoint titrations were conducted, allowing serotype determination in >90% of PUUV- and ANDV-infected patients. Thus, multiparametric IFA enables highly sensitive and specific serological diagnosis of hantavirus infections and can be used to differentiate PUUV and ANDV infection from infections with Murinae-borne hantaviruses (e.g. DOBV and SEOV.

  15. Novel enzyme immunoassay system for simultaneous detection of six subclasses of antiphospholipid antibodies for differential diagnosis of antiphospholipid syndrome.

    Science.gov (United States)

    Nojima, Junzo; Motoki, Yukari; Hara, Kazusa; Sakata, Toshiyuki; Ichihara, Kiyoshi

    2017-06-01

    : Antiphospholipid syndrome, which often complicates systemic lupus erythematosus (SLE), features high occurrence of arterial and/or venous thrombosis and recurrent fetal loss. However, which antibody subclass contributes to which clinical event remains uncertain. We newly developed an up-to-date enzyme immunoassay system using the AcuStar automated analyzer (Instrumentation Laboratory, Bedford, Massachusetts, USA) for parallel detection of six subclasses of antiphospholipid antibodies (aPLs): anticardiolipin antibodies (aCL) of IgG, IgM, and IgA and anti-β2-glycoprotein I antibodies (aβ2GPI) of IgG, IgM, and IgA. They were measured in 276 healthy volunteers and 138 patients with SLE: 45 with thromboembolic complications (29 arterial; 16 venous) and 93 without. Lupus anticoagulant activity in their plasma was measured according to the guidelines recommended by the Subcommittee on Lupus Anticoagulant/Phospholipid-Dependent Antibodies. aCL/β2GPI was measured with a standard ELISA kit commonly used in Japan. The positive results of IgG aCL, IgA aCL, and IgG aβ2GPI were closely associated with thromboembolic complications, whereas IgM aCL and IgM aβ2GPI were not. receiver operating characteristic analysis revealed that the accuracy of predicting thromboembolic complications based on the composite test results of the former three antibodies were obviously higher than by each alone. Regarding agreement with the test results of lupus anticoagulant activity, IgG aβ2GPI showed the closest match. Patients with SLE frequently possess various combinations of the six aPL subclasses, and this antibody spectrum is closely associated with thromboembolic events in these patients. This new automated enzyme immunoassay system allows simultaneous analysis of the profile of aPL subclasses for the differential diagnosis of antiphospholipid antibody syndrome in its early stage.

  16. Simultaneous determination of eight flavonoids in propolis using chemometrics-assisted high performance liquid chromatography-diode array detection.

    Science.gov (United States)

    Sun, Yan-Mei; Wu, Hai-Long; Wang, Jian-Yao; Liu, Zhi; Zhai, Min; Yu, Ru-Qin

    2014-07-01

    A fast analytical strategy of second-order calibration method based on the alternating trilinear decomposition algorithm (ATLD)-assisted high performance liquid chromatography coupled with a diode array detector (HPLC-DAD) was established for the simultaneous determination of eight flavonoids (rutin, quercetin, luteolin, kaempferol, isorhamnetin, apigenin, galangin and chrysin) in propolis capsules samples. The chromatographic separation was implemented on a Wondasil™ C18 column (250mm×4.6mm, 5μm) within 13min with a binary mobile phase composed of water with 1% formic acid and methanol at a flow rate of 1.0mLmin(-1) after flavonoids were only extracted with methanol by ultrasound extraction for 15min. The baseline problem was overcome by considering background drift as additional compositions or factors as well as the target analytes, and ATLD was employed to handle the overlapping peaks from analytes of interest or from analytes and co-eluting matrix compounds. The linearity was good with the correlation coefficients no less than 0.9947; the limit of detections (LODs) within the range of 3.39-33.05ngmL(-1) were low enough; the accuracy was confirmed by the recoveries ranged from 91.9% to 110.2% and the root-mean-square-error of predictions (RMSEPs) less than 1.1μg/mL. The results indicated that the chromatographic method with the aid of ATLD is efficient, sensitive and cost-effective and can realize the resolution and accurate quantification of flavonoids even in the presence of interferences, thus providing an alternative method for accurate quantification of analytes especially when the complete separation is not easily accomplished. The method was successfully applied to propolis capsules samples and the satisfactory results were obtained. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Simultaneous Detection of Key Bacterial Pathogens Related to Pneumonia and Meningitis Using Multiplex PCR Coupled With Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Chi Zhang

    2018-04-01

    Full Text Available Pneumonia and meningitis continue to present an enormous public health burden and pose a major threat to young children. Among the causative organisms of pneumonia and meningitis, bacteria are the most common causes of serious disease and deaths. It is challenging to accurately and rapidly identify these agents. To solve this problem, we developed and validated a 12-plex PCR coupled with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS method (bacterial pathogen-mass spectrometry, BP-MS that can be used to simultaneously screen for 11 key bacterial pathogens related to pneumonia and meningitis. Forty-six nasopharyngeal swabs and 12 isolates were used to determine the specificity of the method. The results showed that, using the BP-MS method, we could accurately identify the expected bacteria without cross-reactivity with other pathogens. For the 11 target bacterial pathogens, the analytical sensitivity of the BP-MS method was as low as 10 copies/reaction. To further evaluate the clinical effectiveness of this method, 204 nasopharyngeal swabs from hospitalized children with suspected pneumonia were tested using this method. In total, 81.9% (167/204 of the samples were positive for at least one of the 11 target pathogens. Among the 167 bacteria-positive samples, the rate of multiple infections was 55.7% (93/167, and the most frequent combination was Streptococcus pneumoniae with Haemophilus influenzae, representing 46.2% (43/93 two-pathogen mixed infections. We used real-time PCR and nested PCR to confirm positive results, with identical results obtained for 81.4% (136/167 of the samples. The BP-MS method is a sensitive and specific molecular detection technique in a multiplex format and with high sample throughput. Therefore, it will be a powerful tool for pathogen screening and antibiotic selection at an early stage of disease.

  18. Simultaneous determination of moxifloxacin and ofloxacin in physiological fluids using high performance liquid chromatography with ultraviolet detection.

    Science.gov (United States)

    Khan, Fahim Ullah; Nasir, Fazli; Iqbal, Zafar; Khan, Ismail; Shahbaz, Naila; Hassan, Muhammad; Ullah, Farhad

    2016-04-01

    A novel, sensitive and validated RP-HPLC-UV method was developed for simultaneous determination of moxifloxacin and ofloxacin using timolol maleate as internal standard in physiological fluids. Different experimental parameters were optimized and validated according to international guidelines. Complete separation of the analytes was achieved with Kromasil 100-5C18 analytical column (250mm×4.6mm×5μm), methanol and 0.05% trifloroacetic acid (TFA) (38:62v/v) were used as mobile phase, pumped at flow rate of 1.1ml/min in isocratic phase, column oven temperature maintained at 45°C and detection wavelength of 290nm. Protein precipitation method was applied to extract the drugs from human plasma and bovine aqueous humor samples using methanol as precipitating solvent. This method is linear in concentration range of 0.018-100μg/ml for moxifloxacin and 0.014-20μg/ml for ofloxacin. The recoveries of the method were 97.52 and 97.39% in human plasma for MX and OFN respectively, while in aqueous humor 94.48% for MX. The LOD values in plasma were found to be 10.0 and 8.00ng/ml for MX and OFN respectively, while their respective LOQ values were 18.0 and 14ng/ml. In aqueous humor the LOD and LOQ for MX were 16.0 and 24ng/ml respectively. In future, this method will be used to study the pharmacokinetic profile of moxifloxacin and ofloxacin in biological fluids and pharmaceutical products. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Chromogenic in situ hybridization is a reliable assay for detection of ALK rearrangements in adenocarcinomas of the lung.

    Science.gov (United States)

    Schildhaus, Hans-Ulrich; Deml, Karl-Friedrich; Schmitz, Katja; Meiboom, Maren; Binot, Elke; Hauke, Sven; Merkelbach-Bruse, Sabine; Büttner, Reinhard

    2013-11-01

    Reliable detection of anaplastic lymphoma kinase (ALK) rearrangements is a prerequisite for personalized treatment of lung cancer patients, as ALK rearrangements represent a predictive biomarker for the therapy with specific tyrosine kinase inhibitors. Currently, fluorescent in situ hybridization (FISH) is considered to be the standard method for assessing formalin-fixed and paraffin-embedded tissue for ALK inversions and translocations. However, FISH requires a specialized equipment, the signals fade rapidly and it is difficult to detect overall morphology and tumor heterogeneity. Chromogenic in situ hybridization (CISH) has been successfully introduced as an alternative test for the detection of several genetic aberrations. This study validates a newly developed ALK CISH assay by comparing FISH and CISH signal patterns in lung cancer samples with and without ALK rearrangements. One hundred adenocarcinomas of the lung were included in this study, among them 17 with known ALK rearrangement. FISH and CISH were carried out and evaluated according to the manufacturers' recommendations. For both assays, tumors were considered positive if ≥15% of tumor cells showed either isolated 3' signals or break-apart patterns or a combination of both. A subset of tumors was exemplarily examined by using a novel EML4 (echinoderm microtubule-associated protein-like 4) CISH probe. Red, green and fusion CISH signals were clearcut and different signal patterns were easily recognized. The percentage of aberrant tumor cells was statistically highly correlated (PCISH. On the basis of 86 samples that were evaluable by ALK CISH, we found a 100% sensitivity and 100% specificity of this assay. Furthermore, EML4 rearrangements could be recognized by CISH. CISH is a highly reliable, sensitive and specific method for the detection of ALK gene rearrangements in pulmonary adenocarcinomas. Our results suggest that CISH might serve as a suitable alternative to FISH, which is the current gold

  20. Linear SVM-Based Android Malware Detection for Reliable IoT Services

    Directory of Open Access Journals (Sweden)

    Hyo-Sik Ham

    2014-01-01

    Full Text Available Current many Internet of Things (IoT services are monitored and controlled through smartphone applications. By combining IoT with smartphones, many convenient IoT services have been provided to users. However, there are adverse underlying effects in such services including invasion of privacy and information leakage. In most cases, mobile devices have become cluttered with important personal user information as various services and contents are provided through them. Accordingly, attackers are expanding the scope of their attacks beyond the existing PC and Internet environment into mobile devices. In this paper, we apply a linear support vector machine (SVM to detect Android malware and compare the malware detection performance of SVM with that of other machine learning classifiers. Through experimental validation, we show that the SVM outperforms other machine learning classifiers.

  1. Autopiquer - a Robust and Reliable Peak Detection Algorithm for Mass Spectrometry.

    Science.gov (United States)

    Kilgour, David P A; Hughes, Sam; Kilgour, Samantha L; Mackay, C Logan; Palmblad, Magnus; Tran, Bao Quoc; Goo, Young Ah; Ernst, Robert K; Clarke, David J; Goodlett, David R

    2017-02-01

    We present a simple algorithm for robust and unsupervised peak detection by determining a noise threshold in isotopically resolved mass spectrometry data. Solving this problem will greatly reduce the subjective and time-consuming manual picking of mass spectral peaks and so will prove beneficial in many research applications. The Autopiquer approach uses autocorrelation to test for the presence of (isotopic) structure in overlapping windows across the spectrum. Within each window, a noise threshold is optimized to remove the most unstructured data, whilst keeping as much of the (isotopic) structure as possible. This algorithm has been successfully demonstrated for both peak detection and spectral compression on data from many different classes of mass spectrometer and for different sample types, and this approach should also be extendible to other types of data that contain regularly spaced discrete peaks. Graphical Abstract ᅟ.

  2. Reliability of panoramic ultrasound imaging in simultaneously examining muscle size and quality of the hamstring muscles in young, healthy males and females.

    Science.gov (United States)

    Palmer, Ty B; Akehi, Kazuma; Thiele, Ryan M; Smith, Doug B; Thompson, Brennan J

    2015-03-01

    The purpose of this study was to examine the reliability of ultrasound (US) measures of cross-sectional area (CSA), muscle thickness (MT) and echo intensity (EI) of the hamstrings, with comparisons between males and females. In 20 healthy participants (10 males, 10 females), CSA, MT and EI were measured from panoramic US scans of the hamstrings on 2 separate days. The intra-class correlation coefficients and standard errors of measurement as a percentage of the mean for CSA, MT and EI ranged from 0.715 to 0.984 and from 3.145 to 12.541% in the males and from 0.724 to 0.977 and from 4.571 to 17.890% in the females, respectively. The males had greater CSAs and MTs and lower EIs than the females (p = 0.002-0.049), and significant relationships were observed between CSA and MT (r = 0.714-0.938, p ≤ 0.001-0.023). From an overall reliability standpoint, these findings suggest that panoramic US may be a reliable technique for examining muscle size and quality of the hamstrings in both males and females. Copyright © 2015 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  3. Clinical accuracy of a PLEX-ID flu device for simultaneous detection and identification of influenza viruses A and B.

    Science.gov (United States)

    Tang, Yi-Wei; Lowery, Kristin S; Valsamakis, Alexandra; Schaefer, Virginia C; Chappell, James D; White-Abell, Jill; Quinn, Criziel D; Li, Haijing; Washington, Cicely A; Cromwell, Jenna; Giamanco, Chantel M; Forman, Michael; Holden, Jeffery; Rothman, Richard E; Parker, Michelle L; Ortenberg, Elaine V; Zhang, Lei; Lin, Yea-Lin; Gaydos, Charlotte A

    2013-01-01

    influenza A virus detection and H1N1-p subtyping. The PLEX-ID Flu assay demonstrated a high level of accuracy for the simultaneous detection and identification of influenza A and B viruses in patient specimens, providing a new laboratory tool for the rapid diagnosis and management of influenza A and B virus infections.

  4. Test-retest reliability and smallest detectable change of the Bristol Impact of Hypermobility (BIoH) questionnaire.

    Science.gov (United States)

    Palmer, S; Manns, S; Cramp, F; Lewis, R; Clark, E M

    2017-12-01

    The Bristol Impact of Hypermobility (BIoH) questionnaire is a patient-reported outcome measure developed in conjunction with adults with Joint Hypermobility Syndrome (JHS). It has demonstrated strong concurrent validity with the Short Form-36 (SF-36) physical component score but other psychometric properties have yet to be established. This study aimed to determine its test-retest reliability and smallest detectable change (SDC). A test-retest reliability study. Participants were recruited from the Hypermobility Syndromes Association, a patient organisation in the United Kingdom. Recruitment packs were sent to 1080 adults who had given permission to be contacted about research. BIoH and SF-36 questionnaires were administered at baseline and repeated two weeks later. An 11-point global rating of change scale (-5 to +5) was also administered at two weeks. Test-retest analysis and calculation of the SDC was conducted on 'stable' patients (defined as global rating of change -1 to +1). 462 responses were received. 233 patients reported a 'stable' condition and were included in analysis (95% women; mean (SD) age 44.5 (13.9) years; BIoH score 223.6 (54.0)). The BIoH questionnaire demonstrated excellent test-retest reliability (ICC 0.923, 95% CI 0.900-0.940). The SDC was 42 points (equivalent to 19% of the mean baseline score). The SF-36 physical and mental component scores demonstrated poorer test-retest reliability and larger SDCs (as a proportion of the mean baseline scores). The results provide further evidence of the potential of the BIoH questionnaire to underpin research and clinical practice for people with JHS. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Test-Retest Reliability and Minimal Detectable Change of the D2 Test of Attention in Patients with Schizophrenia.

    Science.gov (United States)

    Lee, Posen; Lu, Wen-Shian; Liu, Chin-Hsuan; Lin, Hung-Yu; Hsieh, Ching-Lin

    2017-12-08

    The d2 Test of Attention (D2) is a commonly used measure of selective attention for patients with schizophrenia. However, its test-retest reliability and minimal detectable change (MDC) are unknown in patients with schizophrenia, limiting its utility in both clinical and research settings. The aim of the present study was to examine the test-retest reliability and MDC of the D2 in patients with schizophrenia. A rater administered the D2 on 108 patients with schizophrenia twice at a 1-month interval. Test-retest reliability was determined through the calculation of the intra-class correlation coefficient (ICC). We also carried out Bland-Altman analysis, which included a scatter plot of the differences between test and retest against their mean. Systematic biases were evaluated by use of a paired t-test. The ICCs for the D2 ranged from 0.78 to 0.94. The MDCs (MDC%) of the seven subscores were 102.3 (29.7), 19.4 (85.0), 7.2 (94.6), 21.0 (69.0), 104.0 (33.1), 105.0 (35.8), and 7.8 (47.8), which represented limited-to-acceptable random measurement error. Trends in the Bland-Altman plots of the omissions (E1), commissions (E2), and errors (E) were noted, presenting that the data had heteroscedasticity. According to the results, the D2 had good test-retest reliability, especially in the scores of TN, TN-E, and CP. For the further research, finding a way to improve the administration procedure to reduce random measurement error would be important for the E1, E2, E, and FR subscores. © The Author(s) 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. A novel method for rapid and reliable detection of complex vertebral malformation and bovine leukocyte adhesion deficiency in Holstein cattle

    Directory of Open Access Journals (Sweden)

    Zhang Yi

    2012-07-01

    Full Text Available Abstract Background Complex vertebral malformation (CVM and bovine leukocyte adhesion deficiency (BLAD are two autosomal recessive lethal genetic defects frequently occurring in Holstein cattle, identifiable by single nucleotide polymorphisms. The objective of this study is to develop a rapid and reliable genotyping assay to screen the active Holstein sires and determine the carrier frequency of CVM and BLAD in Chinese dairy cattle population. Results We developed real-time PCR-based assays for discrimination of wild-type and defective alleles, so that carriers can be detected. Only one step was required after the DNA extraction from the sample and time consumption was about 2 hours. A total of 587 Chinese Holstein bulls were assayed, and fifty-six CVM-carriers and eight BLAD-carriers were identified, corresponding to heterozygote carrier frequencies of 9.54% and 1.36%, respectively. The pedigree analysis showed that most of the carriers could be traced back to the common ancestry, Osborndale Ivanhoe for BLAD and Pennstate Ivanhoe Star for CVM. Conclusions These results demonstrate that real-time PCR is a simple, rapid and reliable assay for BLAD and CVM defective allele detection. The high frequency of the CVM allele suggests that implementing a routine testing system is necessary to gradually eradicate the deleterious gene from the Chinese Holstein population.

  7. Can magnetic resonance imaging at 3.0-Tesla reliably detect patients with endometriosis? Initial results.

    Science.gov (United States)

    Thomeer, Maarten G; Steensma, Anneke B; van Santbrink, Evert J; Willemssen, Francois E; Wielopolski, Piotr A; Hunink, Myriam G; Spronk, Sandra; Laven, Joop S; Krestin, Gabriel P

    2014-04-01

    The aim of this study was to determine whether an optimized 3.0-Tesla magnetic resonance imaging (MRI) protocol is sensitive and specific enough to detect patients with endometriosis. This was a prospective cohort study with consecutive patients. Forty consecutive patients with clinical suspicion of endometriosis underwent 3.0-Tesla MRI, including a T2-weighted high-resolution fast spin echo sequence (spatial resolution=0.75 ×1.2 ×1.5 mm³) and a 3D T1-weighted high-resolution gradient echo sequence (spatial resolution=0.75 ×1.2 × 2.0 mm³). Two radiologists reviewed the dataset with consensus reading. During laparoscopy, which was used as reference standard, all lesions were characterized according to the revised criteria of the American Fertility Society. Patient-level and region-level sensitivities and specificities and lesion-level sensitivities were calculated. Patient-level sensitivity was 42% for stage I (5/12) and 100% for stages II, III and IV (25/25). Patient-level specificity for all stages was 100% (3/3). The region-level sensitivity and specificity was 63% and 97%, respectively. The sensitivity per lesion was 61% (90% for deep lesions, 48% for superficial lesions and 100% for endometriomata). The detection rate of obliteration of the cul-the-sac was 100% (10/10) with no false positive findings. The interreader agreement was substantial to perfect (kappa=1 per patient, 0.65 per lesion and 0.71 for obliteration of the cul-the-sac). An optimized 3.0-Tesla MRI protocol is accurate in detecting stage II to stage IV endometriosis. © 2014 The Authors. Journal of Obstetrics and Gynaecology Research © 2014 Japan Society of Obstetrics and Gynecology.

  8. Lipase-nanoporous gold biocomposite modified electrode for reliable detection of triglycerides.

    Science.gov (United States)

    Wu, Chao; Liu, Xueying; Li, Yufei; Du, Xiaoyu; Wang, Xia; Xu, Ping

    2014-03-15

    For triglycerides biosensor design, protein immobilization is necessary to create the interface between the enzyme and the electrode. In this study, a glassy carbon electrode (GCE) was modified with lipase-nanoporous gold (NPG) biocomposite (denoted as lipase/NPG/GCE). Due to highly conductive, porous, and biocompatible three-dimensional structure, NPG is suitable for enzyme immobilization. In cyclic voltammetry experiments, the lipase/NPG/GCE bioelectrode displayed surface-confined reaction in a phosphate buffer solution. Linear responses were obtained for tributyrin concentrations ranging from 50 to 250 mg dl(-1) and olive oil concentrations ranging from 10 to 200 mg dl(-1). The value of apparent Michaelis-Menten constant for tributyrin was 10.67 mg dl(-1) and the detection limit was 2.68 mg dl(-1). Further, the lipase/NPG/GCE bioelectrode had strong anti-interference ability against urea, glucose, cholesterol, and uric acid as well as a long shelf-life. For the detection of triglycerides in human serum, the values given by the lipase/NPG/GCE bioelectrode were in good agreement with those of an automatic biochemical analyzer. These properties along with a long self-life make the lipase/NPG/GCE bioelectrode an excellent choice for the construction of triglycerides biosensor. © 2013 Elsevier B.V. All rights reserved.

  9. Delta flow: An accurate, reliable system for detecting kicks and loss of circulation during drilling

    Energy Technology Data Exchange (ETDEWEB)

    Speers, J.M.; Gehrig, G.F.

    1987-12-01

    A system to monitor drilling-fluid flow rate has been developed that detects kicks and lost returns in floating, fixed-platform, and land-base drilling operations. The system uses flowmeters that monitor the flow rates of drilling fluids entering the borehole through the standpipe and leaving the well through the return flowline. These readings are processed in a computer-based, data-acquisition system to form a filtered delta-flow signal that identified the occurrence of downhole fluid gains or losses. The system is designed to trip an alarm when a gain or loss exceeds 25 gal/min (1.6 dm/sup 3//s), even in a floating drilling environment. This sensitivity will generally keep gains or losses to less than 5 bbl (0.8 m/sup 3/).

  10. A rapid and reliable determination of doxycycline hyclate by HPLC with UV detection in pharmaceutical samples

    Directory of Open Access Journals (Sweden)

    SNEZANA S. MITIC

    2008-06-01

    Full Text Available An accurate, sensitive and reproducible high performance liquid chromatographic (HPLC method for the quantification of doxycycline hyclate in pharmaceutical samples has been developed and validated. The drug and the standard were eluted from a Lichrosorb RP-8 (250 mm´4.6 mm, 10 mm particle size at 20 °C with a mobile phase consisting of methanol, acetonitrile and 0.010 M aqueous solution of oxalic acid (2:3:5, v/v/v. The flow rate was 1.25 ml min-1. A UV detector set at 350 nm was used to monitor the effluent. Each analysis required no longer than 4 min. The limits of detection and quantification were 1.15 and 3.84 μg ml-1, respectively. Recoveries for different concentrations ranged from 99.58 to 101.93 %.

  11. Reliability of Using Retinal Vascular Fractal Dimension as a Biomarker in the Diabetic Retinopathy Detection.

    Science.gov (United States)

    Huang, Fan; Dashtbozorg, Behdad; Zhang, Jiong; Bekkers, Erik; Abbasi-Sureshjani, Samaneh; Berendschot, Tos T J M; Ter Haar Romeny, Bart M

    2016-01-01

    The retinal fractal dimension (FD) is a measure of vasculature branching pattern complexity. FD has been considered as a potential biomarker for the detection of several diseases like diabetes and hypertension. However, conflicting findings were found in the reported literature regarding the association between this biomarker and diseases. In this paper, we examine the stability of the FD measurement with respect to (1) different vessel annotations obtained from human observers, (2) automatic segmentation methods, (3) various regions of interest, (4) accuracy of vessel segmentation methods, and (5) different imaging modalities. Our results demonstrate that the relative errors for the measurement of FD are significant and FD varies considerably according to the image quality, modality, and the technique used for measuring it. Automated and semiautomated methods for the measurement of FD are not stable enough, which makes FD a deceptive biomarker in quantitative clinical applications.

  12. Detecting recurrent major depressive disorder within primary care rapidly and reliably using short questionnaire measures.

    Science.gov (United States)

    Thapar, Ajay; Hammerton, Gemma; Collishaw, Stephan; Potter, Robert; Rice, Frances; Harold, Gordon; Craddock, Nicholas; Thapar, Anita; Smith, Daniel J

    2014-01-01

    Major depressive disorder (MDD) is often a chronic disorder with relapses usually detected and managed in primary care using a validated depression symptom questionnaire. However, for individuals with recurrent depression the choice of which questionnaire to use and whether a shorter measure could suffice is not established. To compare the nine-item Patient Health Questionnaire (PHQ-9), the Beck Depression Inventory, and the Hospital Anxiety and Depression Scale against shorter PHQ-derived measures for detecting episodes of DSM-IV major depression in primary care patients with recurrent MDD. Diagnostic accuracy study of adults with recurrent depression in primary care predominantly from Wales Scores on each of the depression questionnaire measures were compared with the results of a semi-structured clinical diagnostic interview using Receiver Operating Characteristic curve analysis for 337 adults with recurrent MDD. Concurrent questionnaire and interview data were available for 272 participants. The one-month prevalence rate of depression was 22.2%. The area under the curve (AUC) and positive predictive value (PPV) at the derived optimal cut-off value for the three longer questionnaires were comparable (AUC = 0.86-0.90, PPV = 49.4-58.4%) but the AUC for the PHQ-9 was significantly greater than for the PHQ-2. However, by supplementing the PHQ-2 score with items on problems concentrating and feeling slowed down or restless, the AUC (0.91) and the PPV (55.3%) were comparable with those for the PHQ-9. A novel four-item PHQ-based questionnaire measure of depression performs equivalently to three longer depression questionnaires in identifying depression relapse in patients with recurrent MDD.

  13. Ultrasensitive and simultaneous detection of hydroquinone, catechol and resorcinol based on the electrochemical co-reduction prepared Au-Pd nanoflower/reduced graphene oxide nanocomposite

    International Nuclear Information System (INIS)

    Chen, Yuan; Liu, Xiaoying; Zhang, Si; Yang, Liuqing; Liu, Meiling; Zhang, Youyu; Yao, Shouzhuo

    2017-01-01

    A simple and efficient eletrochemical sensing platform for simultaneous detection of hydroquinone (HQ), catechol (CC) and resorcinol (RC) based on the Au-Pd bimetallic and graphene is described in this paper. The Au-Pd reduced graphene oxide (Au-Pd NF/rGO) was prepared by the electrochemical co-reduction deposition via cyclic voltammetry method (CV). The Au-Pd NF/rGO nanocomposite was examined by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX) and electrochemical methods CV and differential pulse voltammety (DPV) study showed that the three dihydroxybenzene isomers can be catalytically oxidized and discriminated simultaneously on the Au-Pd NF/rGO/GCE. The presence of Pd makes the performance of the sensor superior to that of in the absence of it. Owing to the integrated superior conductivity and excellent catalytic property of Au-Pd NF/rGO, the sensitive and simultaneous detection of HQ, CC and RC was realized in the individual or triple-components solution based on the as proposed Au-Pd NF/rGO/GCE, which shows wide linear range and low detection limit. The detection of them in tap water, river water and lake water were also successfully performed and good recovery was obtained.

  14. Seismic Azimuthal Anisotropy of the Lower Paleozoic Shales in Northern Poland: can we reliably detect it?

    Science.gov (United States)

    Cyz, Marta; Malinowski, Michał

    2017-04-01

    Analysis of the azimuthal anisotropy is an important aspect of characterization the Lower Paleozoic shale play in northern Poland, since it can be used to map pre-existing fracture networks or help in optimal placement of the horizontal wells. Previous studies employed Velocity versus Azimuth (VVAz) method and found that this anisotropy is weak - on the order of 1-2%, only locally - close to major fault zones - being higher (ca. 7%). This is consistent with the recent re-interpretation of the cross-dipole sonic data, which indicates average shear wave anisotropy of 1%. The problem with the VVAz method is that it requires good definition of the interval, for which the analysis is made and it should be minimum 100 ms thick. In our case, the target intervals are thin - upper reservoir (Lower Silurian Jantar formation) is 15 m thick, lower reservoir (Upper Ordovician Sasino formation) is 25 m thick. Therefore, we prefer to use the Amplitude vs Azimuth (AVAz) method, which can be applied on a single horizon (e.g. the base of the reservoir). However, the AVAz method depends critically on the quality of the seismic data and preservation of amplitudes during processing. On top of the above mentioned issues, physical properties of the Lower Paleozoic shales from Poland seem to be unfavourable for detecting azimuthal anisotropy. For example, for both target formations, parameter g=(Vs/Vp)2 is close to 0.32, which implies that the anisotropy expressed by the anisotropic gradient in the dry (i.e. gas-filled fractures) case is close to zero. In case of e.g. the Bakken Shale, g is much higher (0.38-0.4), leading to a detectable anisotropic signature even in the dry case. Modelling of the synthetic AVAz response performed using available well data suggested that anisotropic gradient in the wet (fluid-filled) case should be detectable even in case of the weak anisotropy (1-2%). This scenario is consistent with the observation, that the studied area is located in the liquid

  15. Detecting inflammation in the unprepared pediatric colon - how reliable is magnetic resonance enterography?

    Energy Technology Data Exchange (ETDEWEB)

    Barber, Joy L.; Watson, Tom A. [Great Ormond Street Hospital for Children NHS Foundation Trust, Department of Radiology, London (United Kingdom); Lozinsky, Adriana Chebar; Kiparissi, Fevronia; Shah, Neil [Great Ormond Street Hospital for Children NHS Foundation Trust, Department of Gastroenterology, London (United Kingdom)

    2016-05-15

    Pediatric inflammatory bowel disease frequently affects the colon. MR enterography is used to assess the small bowel but it also depicts the colon. To compare the accuracy of MR enterography and direct visualization at endoscopy in assessing the colon in pediatric inflammatory bowel disease. We included children with inflammatory bowel disease who had undergone both MR enterography and endoscopy, and we restrospectively assessed the imaging and endoscopic findings. We scored the colonic appearance at MR using a total colon score. We then compared scores for the whole colon and for its individual segments with endoscopy and histology. We included 15 children. An elevated MR colonic segmental score predicted the presence of active inflammation on biopsy with a specificity of 90% (95% confidence interval [CI] 79.5-96.2%) and sensitivity of 60% (CI 40.6-77.3%); this compares reasonably with the predictive values for findings at colonoscopy - specificity 85% (CI 73.4 - 92.9%) and sensitivity 53.3% (CI 34.3%-71.6%). Accuracy did not change significantly with increasing bowel distension. MR-derived scores had comparable accuracy to those derived during visualization at colonoscopy for detecting biopsy-proven inflammation in our patient group. MR enterography might prove useful in guiding biopsy or monitoring treatment response. Collapse of a colonic segment did not impair assessment of inflammation. (orig.)

  16. Can the comet assay be used reliably to detect nanoparticle-induced genotoxicity?

    Science.gov (United States)

    Karlsson, Hanna L; Di Bucchianico, Sebastiano; Collins, Andrew R; Dusinska, Maria

    2015-03-01

    The comet assay is a sensitive method to detect DNA strand breaks as well as oxidatively damaged DNA at the level of single cells. Today the assay is commonly used in nano-genotoxicology. In this review we critically discuss possible interactions between nanoparticles (NPs) and the comet assay. Concerns for such interactions have arisen from the occasional observation of NPs in the "comet head", which implies that NPs may be present while the assay is being performed. This could give rise to false positive or false negative results, depending on the type of comet assay endpoint and NP. For most NPs, an interaction that substantially impacts the comet assay results is unlikely. For photocatalytically active NPs such as TiO2 , on the other hand, exposure to light containing UV can lead to increased DNA damage. Samples should therefore not be exposed to such light. By comparing studies in which both the comet assay and the micronucleus assay have been used, a good consistency between the assays was found in general (69%); consistency was even higher when excluding studies on TiO2 NPs (81%). The strong consistency between the comet and micronucleus assays for a range of different NPs-even though the two tests measure different endpoints-implies that both can be trusted in assessing the genotoxicity of NPs, and that both could be useful in a standard battery of test methods. © 2014 Wiley Periodicals, Inc.

  17. Robust and reliable banknote authentification and print flaw detection with opto-acoustical sensor fusion methods

    Science.gov (United States)

    Lohweg, Volker; Schaede, Johannes; Türke, Thomas

    2006-02-01

    The authenticity checking and inspection of bank notes is a high labour intensive process where traditionally every note on every sheet is inspected manually. However with the advent of more and more sophisticated security features, both visible and invisible, and the requirement of cost reduction in the printing process, it is clear that automation is required. As more and more print techniques and new security features will be established, total quality security, authenticity and bank note printing must be assured. Therefore, this factor necessitates amplification of a sensorial concept in general. We propose a concept for both authenticity checking and inspection methods for pattern recognition and classification for securities and banknotes, which is based on the concept of sensor fusion and fuzzy interpretation of data measures. In the approach different methods of authenticity analysis and print flaw detection are combined, which can be used for vending or sorting machines, as well as for printing machines. Usually only the existence or appearance of colours and their textures are checked by cameras. Our method combines the visible camera images with IR-spectral sensitive sensors, acoustical and other measurements like temperature and pressure of printing machines.

  18. Detecting inflammation in the unprepared pediatric colon - how reliable is magnetic resonance enterography?

    International Nuclear Information System (INIS)

    Barber, Joy L.; Watson, Tom A.; Lozinsky, Adriana Chebar; Kiparissi, Fevronia; Shah, Neil

    2016-01-01

    Pediatric inflammatory bowel disease frequently affects the colon. MR enterography is used to assess the small bowel but it also depicts the colon. To compare the accuracy of MR enterography and direct visualization at endoscopy in assessing the colon in pediatric inflammatory bowel disease. We included children with inflammatory bowel disease who had undergone both MR enterography and endoscopy, and we restrospectively assessed the imaging and endoscopic findings. We scored the colonic appearance at MR using a total colon score. We then compared scores for the whole colon and for its individual segments with endoscopy and histology. We included 15 children. An elevated MR colonic segmental score predicted the presence of active inflammation on biopsy with a specificity of 90% (95% confidence interval [CI] 79.5-96.2%) and sensitivity of 60% (CI 40.6-77.3%); this compares reasonably with the predictive values for findings at colonoscopy - specificity 85% (CI 73.4 - 92.9%) and sensitivity 53.3% (CI 34.3%-71.6%). Accuracy did not change significantly with increasing bowel distension. MR-derived scores had comparable accuracy to those derived during visualization at colonoscopy for detecting biopsy-proven inflammation in our patient group. MR enterography might prove useful in guiding biopsy or monitoring treatment response. Collapse of a colonic segment did not impair assessment of inflammation. (orig.)

  19. A bare-eye-based lateral flow immunoassay based on the use of gold nanoparticles for simultaneous detection of three pesticides

    International Nuclear Information System (INIS)

    Wang, Limin; Cai, Jia; Wang, Yulong; Fang, Qingkui; Wang, Suyan; Cheng, Qi; Liu, Fengquan; Du, Dan; Lin, Yuehe

    2014-01-01

    We present a novel lateral flow immunoassay (LFIA) for the simultaneous detection of the pesticides imidacloprid, chlorpyrifos-methyl and isocarbophos based on three competitive immunoreactions. In contrast to previously reported LFIAs, the method is based on the use of four strips. Each has three red channels (three test lines dispensed with different capture reagent) to detect imidacloprid, chlorpyrifos-methyl and isocarbophos respectively. Different channels on each strip are the key to multi-detection, and four strips of LFIA are needed for visual and semi-quantitative read-outs. Under optimized conditions, the LFIA was applied to the determination of three pesticides. The detection time is within 7 min and the detection limits are 50, 100, and 100 μg L −1 , respectively. Furthermore, the LFIA was applied to the analysis of spiked Chinese cabbage and soil samples and results were validated by HPLC. (author)

  20. Simultaneous Detection of Mixed Infection of Onion yellow dwarf virus and an Allexivirus in RT-PCR for Ensuring Virus Free Onion Bulbs.

    Science.gov (United States)

    Kumar, Sandeep; Baranwal, V K; Joshi, Subodh; Arya, Meenakshi; Majumder, S

    2010-06-01

    Reduced seed production in onion is associated with Onion yellow dwarf virus (OYDV), a filamentous Potyvirus. Onion is also infected with other filamentous virus particles suspected to be Allexivirus. RT-PCR was used to detect mixed infection of both the viruses in leaves and bulbs. A duplex RT-PCR was developed, which simultaneously detected the presence of these two viruses in winter (Rabi) onion bulb. In summer (Kharif) onion bulbs only Allexivirus was detected. The absence of OYDV in summer crop is discussed. The sequencing of RT-PCR amplified products confirmed the identity of OYDV and Allexivirus, the latter showing closer identity to Garlic virus C (GVC)/Garlic mite-borne mosaic virus. This makes the first detection of an Allexivirus in onion crop in India. The duplex RT-PCR to detect these viruses (OYDV and Allexivirus) would be an improvement for indexing of viruses in onion bulbs for seed production.

  1. The sensitivity of computed tomography (CT) scans in detecting trauma: are CT scans reliable enough for courtroom testimony?

    Science.gov (United States)

    Molina, D Kimberley; Nichols, Joanna J; Dimaio, Vincent J M

    2007-09-01

    Rapid and accurate recognition of traumatic injuries is extremely important in emergency room and surgical settings. Emergency departments depend on computed tomography (CT) scans to provide rapid, accurate injury assessment. We conducted an analysis of all traumatic deaths autopsied at the Bexar County Medical Examiner's Office in which perimortem medical imaging (CT scan) was performed to assess the reliability of the CT scan in detecting trauma with sufficient accuracy for courtroom testimony. Cases were included in the study if an autopsy was conducted, a CT scan was performed within 24 hours before death, and there was no surgical intervention. Analysis was performed to assess the correlation between the autopsy and CT scan results. Sensitivity, specificity, positive predictive value, and negative predictive value were defined for the CT scan based on the autopsy results. The sensitivity of the CT scan ranged from 0% for cerebral lacerations, cervical vertebral body fractures, cardiac injury, and hollow viscus injury to 75% for liver injury. This study reveals that CT scans are an inadequate detection tool for forensic pathologists, where a definitive diagnosis is required, because they have a low level of accuracy in detecting traumatic injuries. CT scans may be adequate for clinicians in the emergency room setting, but are inadequate for courtroom testimony. If the evidence of trauma is based solely on CT scan reports, there is a high possibility of erroneous accusations, indictments, and convictions.

  2. A dual-color flow cytometry protocol for the simultaneous detection of Vibrio parahaemolyticus and Salmonella typhimurium using aptamer conjugated quantum dots as labels

    International Nuclear Information System (INIS)

    Duan, Nuo; Wu, Shijia; Yu, Ye; Ma, Xiaoyuan; Xia, Yu; Chen, Xiujuan; Huang, Yukun; Wang, Zhouping

    2013-01-01

    Graphical abstract: -- Highlights: •Two bacteria were simultaneously detected using QD-apt as labels by flow cytometry. •QD-apt were used for recognition and fluorescence detection of two bacteria. •The method was applied successfully for bacteria detection in real samples. -- Abstract: A sensitive, specific method for the collection and detection of pathogenic bacteria was demonstrated using quantum dots (QDs) as a fluorescence marker coupled with aptamers as the molecular recognition element by flow cytometry. The aptamer sequences were selected using a bacterium-based SELEX strategy in our laboratory for Vibrio parahaemolyticus and Salmonella typhimurium that, when applied in this method, allows for the specific recognition of the bacteria from complex mixtures including shrimp samples. Aptamer-modified QDs (QD-apt) were employed to selectively capture and simultaneously detect the target bacteria with high sensitivity using the fluorescence of the labeled QDs. The signal intensity is amplified due to the high photostability of QDs nanoparticles, resulting in improved sensitivity over methods using individual dye-labeled probes. This proposed method is promising for the sensitive detection of other pathogenic bacteria in food stuff if suitable aptamers are chosen. The method may also provide another potential platform for the application of aptamer-conjugated QDs in flow cytometry

  3. A dual-color flow cytometry protocol for the simultaneous detection of Vibrio parahaemolyticus and Salmonella typhimurium using aptamer conjugated quantum dots as labels

    Energy Technology Data Exchange (ETDEWEB)

    Duan, Nuo; Wu, Shijia [State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122 (China); Yu, Ye [Zhangjiagang Entry-Exit Inspection and Quarantine Bureau, Zhangjiangang 215600 (China); Ma, Xiaoyuan; Xia, Yu; Chen, Xiujuan; Huang, Yukun [State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122 (China); Wang, Zhouping, E-mail: wangzp@jiangnan.edu.cn [State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122 (China)

    2013-12-04

    Graphical abstract: -- Highlights: •Two bacteria were simultaneously detected using QD-apt as labels by flow cytometry. •QD-apt were used for recognition and fluorescence detection of two bacteria. •The method was applied successfully for bacteria detection in real samples. -- Abstract: A sensitive, specific method for the collection and detection of pathogenic bacteria was demonstrated using quantum dots (QDs) as a fluorescence marker coupled with aptamers as the molecular recognition element by flow cytometry. The aptamer sequences were selected using a bacterium-based SELEX strategy in our laboratory for Vibrio parahaemolyticus and Salmonella typhimurium that, when applied in this method, allows for the specific recognition of the bacteria from complex mixtures including shrimp samples. Aptamer-modified QDs (QD-apt) were employed to selectively capture and simultaneously detect the target bacteria with high sensitivity using the fluorescence of the labeled QDs. The signal intensity is amplified due to the high photostability of QDs nanoparticles, resulting in improved sensitivity over methods using individual dye-labeled probes. This proposed method is promising for the sensitive detection of other pathogenic bacteria in food stuff if suitable aptamers are chosen. The method may also provide another potential platform for the application of aptamer-conjugated QDs in flow cytometry.

  4. Chlorine activation by N2O5: simultaneous, in situ detection of ClNO2 and N2O5 by chemical ionization mass spectrometry

    Directory of Open Access Journals (Sweden)

    J. A. Thornton

    2009-05-01

    Full Text Available We report a new method for the simultaneous in situ detection of nitryl chloride (ClNO2 and dinitrogen pentoxide (N2O5 using chemical ionization mass spectrometry (CIMS. The technique relies on the formation and detection of iodide ion-molecule clusters, I(ClNO2− and I(N2O5−. The novel N2O5 detection scheme is direct. It does not suffer from high and variable chemical interferences, which are associated with the typical method of nitrate anion detection. We address the role of water vapor, CDC electric field strength, and instrument zero determinations, which influence the overall sensitivity and detection limit of this method. For both species, the method demonstrates high sensitivity (>1 Hz/pptv, precision (~10% for 100 pptv in 1 s, and accuracy (~20%, the latter ultimately determined by the nitrogen dioxide (NO2 cylinder calibration standard and characterization of inlet effects. For the typically low background signals (S/N ratios of 2 for 1 pptv in 60 s averages, but uncertainty associated with the instrumental zero currently leads to an ultimate detection limit of ~5 pptv for both species. We validate our approach for the simultaneous in situ measurement of ClNO2 and N2O5 while on board the R/V Knorr as part of the ICEALOT 2008 Field Campaign.

  5. A Targeted LC-MS/MS Method for the Simultaneous Detection and Quantitation of Egg, Milk, and Peanut Allergens in Sugar Cookies.

    Science.gov (United States)

    Boo, Chelsea C; Parker, Christine H; Jackson, Lauren S

    2018-01-01

    Food allergy is a growing public health concern, with many individuals reporting allergies to multiple food sources. Compliance with food labeling regulations and prevention of inadvertent cross-contact in manufacturing requires the use of reliable methods for the detection and quantitation of allergens in processed foods. In this work, a novel liquid chromatography-tandem mass spectrometry multiple-reaction monitoring method for multiallergen detection and quantitation of egg, milk, and peanut was developed and evaluated in an allergen-incurred baked sugar cookie matrix. A systematic evaluation of method parameters, including sample extraction, concentration, and digestion, were optimized for candidate allergen peptide markers. The optimized method enabled the reliable detection and quantitation of egg, milk, and peanut allergens in sugar cookies, with allergen concentrations as low as 5 ppm allergen-incurred ingredient.

  6. Water-soluble upper GI based on clinical findings is reliable to detect anastomotic leaks after laparoscopic gastric bypass.

    Science.gov (United States)

    Katasani, V G; Leeth, R R; Tishler, D S; Leath, T D; Roy, B P; Canon, C L; Vickers, S M; Clements, R H

    2005-11-01

    Anastomotic leak after laparoscopic Roux-en-Y gastric bypass (LGB) is a major complication that must be recognized and treated early for best results. There is controversy in the literature regarding the reliability of upper GI series (UGI) in diagnosing leaks. LGB was performed in patients meeting NIH criteria for the surgical treatment of morbid obesity. All leaks identified at the time of surgery were repaired with suture and retested. Drains were placed at the surgeon's discretion. Postoperatively, UGI was performed by an experienced radiologist if there was a clinical suspicion of leak. From September 2001 until October 2004, a total of 553 patients (age 40.4 +/- 9.2 years, BMI 48.6 +/- 7.2) underwent LGB at UAB. Seventy-eight per cent (431 of 553) of patients had no clinical evidence suggesting anastomotic leak and were managed expectantly. Twenty-two per cent (122 of 553) of patients met at least one inclusion criteria for leak and underwent UGI. Four of 122 patients (3.2%) had a leak, two from anastomosis and two from the perforation of the stapled end of the Roux limb. No patient returned to the operating room without a positive UGI. High clinical suspicion and selectively performed UGI based on clinical evidence is reliable in detecting leaks.

  7. Usefulness of the dynamic gadolinium-enhanced magnetic resonance imaging with simultaneous acquisition of coronal and sagittal planes for detection of pituitary microadenomas.

    Science.gov (United States)

    Lee, Han Bee; Kim, Sung Tae; Kim, Hyung-Jin; Kim, Keon Ha; Jeon, Pyoung; Byun, Hong Sik; Choi, Jin Wook

    2012-03-01

    Does dynamic gadolinium-enhanced imaging with simultaneous acquisition of coronal and sagittal planes improve diagnostic accuracy of pituitary microadenomas compared with coronal images alone? Fifty-six patients underwent 3-T sella MRI including dynamic simultaneous acquisition of coronal and sagittal planes after gadolinium injection. According to conspicuity, lesions were divided into four scores (0, no; 1, possible; 2, probable; 3, definite delayed enhancing lesion). Additional information on supplementary sagittal images compared with coronal ones was evaluated with a 4-point score (0, no; 1, possible; 2, probable; 3, definite additional information). Accuracy of tumour detection was calculated. Average scores for lesion detection of a combination of two planes, coronal, and sagittal images were 2.59, 2.32, and 2.18. 6/10 lesions negative on coronal images were detected on sagittal ones. Accuracy of a combination of two planes, of coronal and of sagittal images was 92.86%, 82.14% and 75%. Six patients had probable or definite additional information on supplementary sagittal images compared with coronal ones alone (10.71%). Dynamic MRI with combined coronal and sagittal planes was more accurate for detection of pituitary microadenomas than routinely used coronal images. Simultaneous dynamic enhanced acquisition can make study time fast and costs low. We present a new dynamic MRI technique for evaluating pituitary microadenomas • This technique provides simultaneous acquisition of contrast enhanced coronal and sagittal images. • This technique makes the diagnosis more accurate and reduces the examination time. • Such MR imaging only requires one single bolus of contrast agent.

  8. Simultaneous detection of genetically modified organisms by multiplex ligation-dependent genome amplification and capillary gel electrophoresis with laser-induced fluorescence.

    Science.gov (United States)

    García-Cañas, Virginia; Mondello, Monica; Cifuentes, Alejandro

    2010-07-01

    In this work, an innovative method useful to simultaneously analyze multiple genetically modified organisms is described. The developed method consists in the combination of multiplex ligation-dependent genome dependent amplification (MLGA) with CGE and LIF detection using bare-fused silica capillaries. The MLGA process is based on oligonucleotide constructs, formed by a universal sequence (vector) and long specific oligonucleotides (selectors) that facilitate the circularization of specific DNA target regions. Subsequently, the circularized target sequences are simultaneously amplified with the same couple of primers and analyzed by CGE-LIF using a bare-fused silica capillary and a run electrolyte containing 2-hydroxyethyl cellulose acting as both sieving matrix and dynamic capillary coating. CGE-LIF is shown to be very useful and informative for optimizing MLGA parameters such as annealing temperature, number of ligation cycles, and selector probes concentration. We demonstrate the specificity of the method in detecting the presence of transgenic DNA in certified reference and raw commercial samples. The method developed is sensitive and allows the simultaneous detection in a single run of percentages of transgenic maize as low as 1% of GA21, 1% of MON863, and 1% of MON810 in maize samples with signal-to-noise ratios for the corresponding DNA peaks of 15, 12, and 26, respectively. These results demonstrate, to our knowledge for the first time, the great possibilities of MLGA techniques for genetically modified organisms analysis.

  9. Simultaneous determination of chromium(III) and chromium(VI) in aqueous solutions by ion chromatography and chemiluminescence detection

    DEFF Research Database (Denmark)

    Gammelgaard, Bente; Jøns, O; Nielsen, B

    1992-01-01

    A method for the simultaneous determination of chromium(iii) and chromium(vi) in a flow system based on chemiluminescence was developed. A Dionex cation-exchange guard column was used to separate chromium(iii) from chromium(vi), and chromium(vi) was reduced by potassium sulfite, whereupon both...

  10. Separation and Simultaneous Determination of 14 Fungicides with the Combination of Multi-Analyte Methods and HPLC Detection

    Energy Technology Data Exchange (ETDEWEB)

    Canping, Pan [Department of Applied Chemistry, China Agricultural University, Beijing (China)

    2009-07-15

    The separation and simultaneous HPLC-MS determination for a series of fungicide products is reported. Multi-analyte methods were applied on a Chromolith RP-18e monolithic column having low resistance and enabling high flow rates and short analysis time at very good separation power. Details and analytical conditions are described with chromatograms illustrating the results and work done. (author)

  11. Simultaneous determination of ammonia, dimethylamine, trimethylamine and trimethylamine-N-oxide in fish extracts by capillary electrophoresis with indirect UV-detection

    DEFF Research Database (Denmark)

    Timm Heinrich, Maike; Jørgensen, Bo

    2002-01-01

    A capillary electrophoretic method with indirect UV detection is described for simultaneous determination of ammonia, dimethylamine (DMA), trimethylamine (TMA) and trimethylamine-N- oxide (TMAO) in aqueous extracts of fish, A buffer consisting of 4 mM formic acid, 5 mM copper(II)sulfate and 3 m......M. The detection limit for ammonia, DMA, TMA, and TMAO was less than 0.04 mM, corresponding to 2 mg nitrogen per 100 g fish. As an extra benefit, the method also provided a quantitative determination of potassium, sodium, calcium and magnesium ions. (C) 2002 Elsevier Science Ltd. All rights reserved....

  12. Reliability of the MicroScan WalkAway PC21 panel in identifying and detecting oxacillin resistance in clinical coagulase-negative staphylococci strains.

    Science.gov (United States)

    Olendzki, A N; Barros, E M; Laport, M S; Dos Santos, K R N; Giambiagi-Demarval, M

    2014-01-01

    The purpose of this study was to determine the reliability of the MicroScan WalkAway PosCombo21 (PC21) system for the identification of coagulase-negative staphylococci (CNS) strains and the detection of oxacillin resistance. Using molecular and phenotypic methods, 196 clinical strains were evaluated. The automated system demonstrated 100 % reliability for the identification of the clinical strains Staphylococcus haemolyticus, Staphylococcus hominis and Staphylococcus cohnii; 98.03 % reliability for the identification of Staphylococcus epidermidis; 70 % reliability for the identification of Staphylococcus lugdunensis; 40 % reliability for the identification of Staphylococcus warneri; and 28.57 % reliability for the identification of Staphylococcus capitis, but no reliability for the identification of Staphylococcus auricularis, Staphylococcus simulans and Staphylococcus xylosus. We concluded that the automated system provides accurate results for the more common CNS species but often fails to accurately identify less prevalent species. For the detection of oxacillin resistance, the automated system showed 100 % specificity and 90.22 % sensitivity. Thus, the PC21 panel detects oxacillin-resistant strains, but is limited by the heteroresistance that is observed when using most phenotypic methods.

  13. Electrochemical Selective and Simultaneous Detection of Diclofenac and Ibuprofen in Aqueous Solution Using HKUST-1 Metal-Organic Framework-Carbon Nanofiber Composite Electrode

    Directory of Open Access Journals (Sweden)

    Sorina Motoc

    2016-10-01

    Full Text Available In this study, the detection protocols for the individual, selective, and simultaneous determination of ibuprofen (IBP and diclofenac (DCF in aqueous solutions have been developed using HKUST-1 metal-organic framework-carbon nanofiber composite (HKUST-CNF electrode. The morphological and electrical characterization of modified composite electrode prepared by film casting was studied by scanning electronic microscopy and four-point-probe methods. The electrochemical characterization of the electrode by cyclic voltammetry (CV was considered the reference basis for the optimization of the operating conditions for chronoamperometry (CA and multiple-pulsed amperometry (MPA. This electrode exhibited the possibility to selectively detect IBP and DCF by simple switching the detection potential using CA. However, the MPA operated under optimum working conditions of four potential levels selected based on CV shape in relation to the potential value, pulse time, and potential level number, and order allowed the selective/simultaneous detection of IBP and DCF characterized by the enhanced detection performance. For this application, the HKUST-CNF electrode exhibited a good stability and reproducibility of the results was achieved.

  14. Electrochemical Selective and Simultaneous Detection of Diclofenac and Ibuprofen in Aqueous Solution Using HKUST-1 Metal-Organic Framework-Carbon Nanofiber Composite Electrode.

    Science.gov (United States)

    Motoc, Sorina; Manea, Florica; Iacob, Adriana; Martinez-Joaristi, Alberto; Gascon, Jorge; Pop, Aniela; Schoonman, Joop

    2016-10-17

    In this study, the detection protocols for the individual, selective, and simultaneous determination of ibuprofen (IBP) and diclofenac (DCF) in aqueous solutions have been developed using HKUST-1 metal-organic framework-carbon nanofiber composite (HKUST-CNF) electrode. The morphological and electrical characterization of modified composite electrode prepared by film casting was studied by scanning electronic microscopy and four-point-probe methods. The electrochemical characterization of the electrode by cyclic voltammetry (CV) was considered the reference basis for the optimization of the operating conditions for chronoamperometry (CA) and multiple-pulsed amperometry (MPA). This electrode exhibited the possibility to selectively detect IBP and DCF by simple switching the detection potential using CA. However, the MPA operated under optimum working conditions of four potential levels selected based on CV shape in relation to the potential value, pulse time, and potential level number, and order allowed the selective/simultaneous detection of IBP and DCF characterized by the enhanced detection performance. For this application, the HKUST-CNF electrode exhibited a good stability and reproducibility of the results was achieved.

  15. Simultaneous detection of viruses and Toxoplasma gondii in cerebrospinal fluid specimens by multiplex polymerase chain reaction-based reverse hybridization assay.

    Science.gov (United States)

    Del Prete, Raffaele; Di Taranto, Anna Maria; Lipsi, Maria Rosaria; Natalicchio, Maria Iole; Antonetti, Raffaele; Miragliotta, Giuseppe

    2009-04-01

    The lack of rapidity and the low sensitivity and specificity of traditional laboratory methods limits their usefulness in the laboratory diagnosis of viral central nervous system (CNS) infections. This study describes the use of a commercially available multiplex polymerase chain reaction (mPCR)-based reverse hybridization assay (RHA) for the simultaneous detection of the genomes of 8 viruses and Toxoplasma gondii in cerebrospinal fluids (CSF) from 181 patients suspected of having viral meningitis. Twenty-two/181 (12.15%) CSF samples resulted positive by mPCR. Eighteen/22 were positive for 1 viral pathogen, whereas a dual infection was detected in 4/22 samples. Epstein-Barr virus (EBV) was the most commonly detected virus (6/22), followed by herpes simplex virus type-1 (HSV-1) (5/22) and -2 (HSV-2) (4/22). Cytomegalovirus (CMV), human herpesvirus-6 (HHV-6), and Epstein-Barr virus (EBV) were detected in 1 specimen each. Two CSF samples were co-infected by HSV-1/HSV-2, 1 sample by HHV-6/T. gondii, and 1 sample by EBV/EV, respectively. Our data support the usefulness of mPCR as a rapid molecular method for the simultaneous detection of major viral pathogens and T. gondii in aseptic meningitis also to allow the earlier application of specific antiviral therapy.

  16. Simultaneous Determination of Bergapten, Imperatorin, Notopterol, and Isoimperatorin in Rat Plasma by High Performance Liquid Chromatography with Fluorescence Detection and Its Application to Pharmacokinetic and Excretion Study after Oral Administration of Notopterygium incisum Extract

    Directory of Open Access Journals (Sweden)

    John Teye Azietaku

    2016-01-01

    Full Text Available A specific, sensitive, and reliable high performance liquid chromatography with fluorescence detection (HPLC-FLD was first optimized and then used in the simultaneous quantification of bergapten, imperatorin, notopterol, and isoimperatorin in rat plasma using osthole as the internal standard. Liquid-liquid extraction with ethyl acetate was employed in treating the rat plasma samples obtained. Separation was carried out with a Hedera™ ODS column (4.6 × 250 mm, 5 μm by gradient elution at a temperature of 40°C. Excitation and emission of the fluorescence detector were set to 300 and 490 nm, respectively. The lower limits of quantification for bergapten, imperatorin, notopterol, and isoimperatorin in rat plasma were 4, 40, 4, and 2 ng mL−1, respectively. The intraday and interday precision and accuracy for the four coumarins were within acceptable criteria. The recovery of the method was satisfactory with a range of 80.3–114%. The validated method was successfully used for the simultaneous determination of the four coumarins in Notopterygium incisum extracts and also for the pharmacokinetic and excretion study of bergapten, imperatorin, notopterol, and isoimperatorin in rats.

  17. Ambient Pressure Laser Desorption—Chemical Ionization Mass Spectrometry for Fast and Reliable Detection of Explosives, Drugs, and Their Precursors

    Directory of Open Access Journals (Sweden)

    René Reiss

    2018-06-01

    Full Text Available Fast and reliable information is crucial for first responders to draw correct conclusions at crime scenes. An ambient pressure laser desorption (APLD mass spectrometer is introduced for this scenario, which enables detecting substances on surfaces without sample pretreatment. It is especially useful for substances with low vapor pressure and thermolabile ones. The APLD allows for the separation of desorption and ionization into two steps and, therefore, both can be optimized separately. Within this work, an improved version of the developed system is shown that achieves limits of detection (LOD down to 500 pg while remaining fast and flexible. Furthermore, realistic scenarios are applied to prove the usability of this system in real-world issues. For this purpose, post-blast residues of a bomb from the Second World War were analyzed, and the presence of PETN was proven without sample pretreatment. In addition, the analyzable substance range could be expanded by various drugs and drug precursors. Thus, the presented instrumentation can be utilized for an increased number of forensically important compound classes without changing the setup. Drug precursors revealed a LOD ranging from 6 to 100 ng. Drugs such as cocaine hydrochloride, heroin, (3,4-methylendioxy-methamphetamine hydrochloride (MDMA hydrochloride, and others exhibit a LOD between 10 to 200 ng.

  18. Reliability and relative validity of a child nutrition questionnaire to simultaneously assess dietary patterns associated with positive energy balance and food behaviours, attitudes, knowledge and environments associated with healthy eating

    Directory of Open Access Journals (Sweden)

    Magarey Anthea M

    2008-01-01

    Full Text Available Abstract Background Food behaviours, attitudes, environments and knowledge are relevant to professionals in childhood obesity prevention, as are dietary patterns which promote positive energy balance. There is a lack of valid and reliable tools to measure these parameters. The aim of this study was to determine the reliability and relative validity of a child nutrition questionnaire assessing all of these parameters, used in the evaluation of a community-based childhood obesity prevention project. Methods The development of the 14-item questionnaire was informed by the aims of the obesity prevention project. A sub-sample of children aged 10–12 years from primary schools involved in the intervention was recruited at the project's baseline data collection (Test 1. Questionnaires were readministered (Test 2 following which students completed a 7-day food diary designed to reflect the questionnaire. Twelve scores were derived to assess consumption of fruit, vegetables, water, noncore foods and sweetened beverages plus food knowledge, behaviours, attitudes and environments. Reliability was assessed using (a the intra class correlation coefficient (ICC and 95% confidence intervals to compare scores from Tests 1 and 2 (test-retest reliability and (b Cronbach's alpha (internal consistency. Validity was assessed with Spearman correlations, bias and limits of agreement between scores from Test 1 and the 7-day diaries. The Wilcoxon signed rank test checked for significant differences between mean scores. Results One hundred and forty one students consented to the study. Test 2 (n = 134 occurred between eight and 36 days after Test 1. For 10/12 scores ICCs ranged from 0.47–0.66 (p 0.05 for 10/12 (test-retest reliability and 3/7 (validity scores. Conclusion This child nutrition questionnaire is a valid and reliable tool to simultaneously assess dietary patterns associated with positive energy balance, and food behaviours, attitudes and environments in

  19. Is air-displacement plethysmography a reliable method of detecting ongoing changes in percent body fat within obese children involved in a weight management program?

    DEFF Research Database (Denmark)

    Ewane, Cecile; McConkey, Stacy A; Kreiter, Clarence D

    2010-01-01

    (percent body fat) over time. The gold standard method, hydrodensitometry, has severe limitations for the pediatric population. OBJECTIVE: This study examines the reliability of air-displacement plethysmography (ADP) in detecting percent body fat changes within obese children over time. METHODS: Percent...... body fat by ADP, weight, and body mass index (BMI) were measured for eight obese children aged 5-12 years enrolled in a weight management program over a 12-month period. These measurements were taken at initial evaluation, 1.5 months, 3 months, 6 months, and 12 months to monitor the progress...... of the subjects and detect any changes in these measures over time. Statistical analysis was used to determine the reliability of the data collected. RESULTS: The reliability estimate for percent body fat by ADP was 0.78. This was much lower than the reliability of BMI, 0.98, and weight measurements, 0...

  20. 3-lead electrocardiogram is more reliable than pulse oximetry to detect bradycardia during stabilisation at birth of very preterm infants.

    Science.gov (United States)

    Iglesias, Beatriz; Rodrí Guez, Marí A José; Aleo, Esther; Criado, Enrique; Martí Nez-Orgado, Jose; Arruza, Luis

    2018-05-01

    Current neonatal resuscitation guidelines suggest the use of ECG in the delivery room (DR) to assess heart rate (HR). However, reliability of ECG compared with pulse oximetry (PO) in a situation of bradycardia has not been specifically investigated. The objective of the present study was to compare HR monitoring using ECG or PO in a situation of bradycardia (HR <100 beats per minute (bpm)) during preterm stabilisation in the DR. Video recordings of resuscitations of infants <32 weeks of gestation were reviewed. HR readings in a situation of bradycardia (<100 bpm) at any moment during stabilisation were registered with both devices every 5 s from birth. A total of 29 episodes of bradycardia registered by the ECG in 39 video recordings were included in the analysis (n=29). PO did not detect the start of these events in 20 cases (69%). PO detected the start and the end of bradycardia later than the ECG (median (IQR): 5 s (0-10) and 5 s (0-7.5), respectively). A decline in PO accuracy was observed as bradycardia progressed so that by the end of the episode PO offered significantly lower HR readings than ECG. PO detects the start and recovery of bradycardia events slower and less accurately than ECG during stabilisation at birth of very preterm infants. ECG use in this scenario may contribute to an earlier initiation of resuscitation manoeuvres and to avoid unnecessary prolongation of resuscitation efforts after recovery. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  1. The Space-Time Variation of Global Crop Yields, Detecting Simultaneous Outliers and Identifying the Teleconnections with Climatic Patterns

    Science.gov (United States)

    Najafi, E.; Devineni, N.; Pal, I.; Khanbilvardi, R.

    2017-12-01

    An understanding of the climate factors that influence the space-time variability of crop yields is important for food security purposes and can help us predict global food availability. In this study, we address how the crop yield trends of countries globally were related to each other during the last several decades and the main climatic variables that triggered high/low crop yields simultaneously across the world. Robust Principal Component Analysis (rPCA) is used to identify the primary modes of variation in wheat, maize, sorghum, rice, soybeans, and barley yields. Relations between these modes of variability and important climatic variables, especially anomalous sea surface temperature (SSTa), are examined from 1964 to 2010. rPCA is also used to identify simultaneous outliers in each year, i.e. systematic high/low crop yields across the globe. The results demonstrated spatiotemporal patterns of these crop yields and the climate-related events that caused them as well as the connection of outliers with weather extremes. We find that among climatic variables, SST has had the most impact on creating simultaneous crop yields variability and yield outliers in many countries. An understanding of this phenomenon can benefit global crop trade networks.

  2. One-step simultaneous differential scanning calorimetry-FTIR microspectroscopy to quickly detect continuous pathways in the solid-state glucose/asparagine Maillard reaction.

    Science.gov (United States)

    Hwang, Deng-Fwu; Hsieh, Tzu-Feng; Lin, Shan-Yang

    2013-01-01

    The stepwise reaction pathway of the solid-state Maillard reaction between glucose (Glc) and asparagine (Asn) was investigated using simultaneous differential scanning calorimetry (DSC)-FTIR microspectroscopy. The color change and FTIR spectra of Glc-Asn physical mixtures (molar ratio = 1:1) preheated to different temperatures followed by cooling were also examined. The successive reaction products such as Schiff base intermediate, Amadori product, and decarboxylated Amadori product in the solid-state Glc-Asn Maillard reaction were first simultaneously evidenced by this unique DSC-FTIR microspectroscopy. The color changed from white to yellow-brown to dark brown, and appearance of new IR peaks confirmed the formation of Maillard reaction products. The present study clearly indicates that this unique DSC-FTIR technique not only accelerates but also detects precursors and products of the Maillard reaction in real time.

  3. Reliability engineering

    International Nuclear Information System (INIS)

    Lee, Chi Woo; Kim, Sun Jin; Lee, Seung Woo; Jeong, Sang Yeong

    1993-08-01

    This book start what is reliability? such as origin of reliability problems, definition of reliability and reliability and use of reliability. It also deals with probability and calculation of reliability, reliability function and failure rate, probability distribution of reliability, assumption of MTBF, process of probability distribution, down time, maintainability and availability, break down maintenance and preventive maintenance design of reliability, design of reliability for prediction and statistics, reliability test, reliability data and design and management of reliability.

  4. Atrial fibrillation detected by external loop recording for seven days or two-day simultaneous Holter recording: A comparison in patients with ischemic stroke or transient ischemic attack.

    Science.gov (United States)

    Sejr, Michala Herskind; Nielsen, Jens Cosedis; Damgaard, Dorte; Sandal, Birgitte Forsom; May, Ole

    Atrial fibrillation (AF) is the most common cardiac cause of ischemic stroke and transient ischemic attack (IS/TIA). To compare the diagnostic value of seven-day external loop recording (ELR) and two-day Holter recording for detecting AF after IS/TIA. 191 IS/TIA patients without AF history. Endpoint was AF >30s. We started two-day Holter recording and seven-day ELR simultaneously. Seven-day ELR and two-day Holter recording detected the same three AF patients. ELR detected another six patients with AF adjudicated by cardiologists, four detections after Holter (3 vs. 7, p=0.125) and two false-positive detections during Holter. Seven-day ELR automatically classified 50/191 patients (26%) with AF, but only 7/50 (14%) were confirmed as AF by cardiologists. Seven-day ELR did not detect significantly more patients with AF than two-day Holter recording. 86% of patients with ELR-classified AF were false positives, indicating a poor performance of the automatic AF detection algorithm used. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. 1H-detected MAS solid-state NMR experiments enable the simultaneous mapping of rigid and dynamic domains of membrane proteins

    Science.gov (United States)

    Gopinath, T.; Nelson, Sarah E. D.; Veglia, Gianluigi

    2017-12-01

    Magic angle spinning (MAS) solid-state NMR (ssNMR) spectroscopy is emerging as a unique method for the atomic resolution structure determination of native membrane proteins in lipid bilayers. Although 13C-detected ssNMR experiments continue to play a major role, recent technological developments have made it possible to carry out 1H-detected experiments, boosting both sensitivity and resolution. Here, we describe a new set of 1H-detected hybrid pulse sequences that combine through-bond and through-space correlation elements into single experiments, enabling the simultaneous detection of rigid and dynamic domains of membrane proteins. As proof-of-principle, we applied these new pulse sequences to the membrane protein phospholamban (PLN) reconstituted in lipid bilayers under moderate MAS conditions. The cross-polarization (CP) based elements enabled the detection of the relatively immobile residues of PLN in the transmembrane domain using through-space correlations; whereas the most dynamic region, which is in equilibrium between folded and unfolded states, was mapped by through-bond INEPT-based elements. These new 1H-detected experiments will enable one to detect not only the most populated (ground) states of biomacromolecules, but also sparsely populated high-energy (excited) states for a complete characterization of protein free energy landscapes.

  6. Detecting concealed information from groups using a dynamic questioning approach: simultaneous skin conductance measurement and immediate feedback

    NARCIS (Netherlands)

    Meijer, E.H.; Bente, G.; Ben-Shakhar, G.; Schumacher, A.

    2013-01-01

    Lie detection procedures typically aim at determining the guilt or innocence of a single suspect. The Concealed Information Test (CIT), for example, has been shown to be highly successful in detecting the presence or absence of crime-related information in a suspect's memory. Many of today's

  7. Microarray multiplex assay for the simultaneous detection and discrimination of hepatitis B, hepatitis C, and human immunodeficiency type-1 viruses in human blood samples

    International Nuclear Information System (INIS)

    Hsia, Chu Chieh; Chizhikov, Vladimir E.; Yang, Amy X.; Selvapandiyan, Angamuthu; Hewlett, Indira; Duncan, Robert; Puri, Raj K.; Nakhasi, Hira L.; Kaplan, Gerardo G.

    2007-01-01

    Hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type-1 (HIV-1) are transfusion-transmitted human pathogens that have a major impact on blood safety and public health worldwide. We developed a microarray multiplex assay for the simultaneous detection and discrimination of these three viruses. The microarray consists of 16 oligonucleotide probes, immobilized on a silylated glass slide. Amplicons from multiplex PCR were labeled with Cy-5 and hybridized to the microarray. The assay detected 1 International Unit (IU), 10 IU, 20 IU of HBV, HCV, and HIV-1, respectively, in a single multiplex reaction. The assay also detected and discriminated the presence of two or three of these viruses in a single sample. Our data represent a proof-of-concept for the possible use of highly sensitive multiplex microarray assay to screen and confirm the presence of these viruses in blood donors and patients

  8. Reliable allele detection using SNP-based PCR primers containing Locked Nucleic Acid: application in genetic mapping

    Directory of Open Access Journals (Sweden)

    Trognitz Friederike

    2007-02-01

    Full Text Available Abstract Background The diploid, Solanum caripense, a wild relative of potato and tomato, possesses valuable resistance to potato late blight and we are interested in the genetic base of this resistance. Due to extremely low levels of genetic variation within the S. caripense genome it proved impossible to generate a dense genetic map and to assign individual Solanum chromosomes through the use of conventional chromosome-specific SSR, RFLP, AFLP, as well as gene- or locus-specific markers. The ease of detection of DNA polymorphisms depends on both frequency and form of sequence variation. The narrow genetic background of close relatives and inbreds complicates the detection of persisting, reduced polymorphism and is a challenge to the development of reliable molecular markers. Nonetheless, monomorphic DNA fragments representing not directly usable conventional markers can contain considerable variation at the level of single nucleotide polymorphisms (SNPs. This can be used for the design of allele-specific molecular markers. The reproducible detection of allele-specific markers based on SNPs has been a technical challenge. Results We present a fast and cost-effective protocol for the detection of allele-specific SNPs by applying Sequence Polymorphism-Derived (SPD markers. These markers proved highly efficient for fingerprinting of individuals possessing a homogeneous genetic background. SPD markers are obtained from within non-informative, conventional molecular marker fragments that are screened for SNPs to design allele-specific PCR primers. The method makes use of primers containing a single, 3'-terminal Locked Nucleic Acid (LNA base. We demonstrate the applicability of the technique by successful genetic mapping of allele-specific SNP markers derived from monomorphic Conserved Ortholog Set II (COSII markers mapped to Solanum chromosomes, in S. caripense. By using SPD markers it was possible for the first time to map the S. caripense alleles

  9. Mammographic casting-type calcification associated with small screen-detected invasive breast cancers: is this a reliable prognostic indicator?

    International Nuclear Information System (INIS)

    Peacock, C.; Given-Wilson, R.M.; Duffy, S.W.

    2004-01-01

    AIM: The aim of the present study was to establish whether mammographic casting-type calcification associated with small screen-detected invasive breast cancers is a reliable prognostic indicator. METHODS AND MATERIALS: We retrospectively identified 50 consecutive women diagnosed with an invasive cancer less than 15 mm who showed associated casting calcification on their screening mammograms. Controls were identified that showed no microcalcification and were matched for tumour size, histological type and lymph node status. A minimum of 5 years follow-up was obtained, noting recurrence and outcome. Conditional and unconditional logistic regression, depending on the outcome variable, were used to analyse the data, taking the matched design into account in both cases. Where small numbers prohibited the use of logistic regression, Fisher's exact test was used. RESULTS: Five deaths from breast cancer occurred out of the 50 cases, of which three were lymph node positive, two were lymph node negative and none were grade 3. None of the 78 control cases died from breast cancer. The difference in breast cancer death rates was significant by Fisher's exact test (p=0.02). Risk of recurrence was also significantly increased in the casting cases (OR=3.55, 95% CI 1.02-12.33, p=0.046). CONCLUSION: Although the overall outcome for small screen-detected breast cancers is good, our study suggests that casting calcification is a poorer prognostic factor. The advantage of a mammographic feature as an independent prognostic indicator lies in early identification of high-risk patients, allowing optimization of management

  10. Sampling inspection for the evaluation of time-dependent reliability of deteriorating systems under imperfect defect detection

    International Nuclear Information System (INIS)

    Kuniewski, Sebastian P.; Weide, Johannes A.M. van der; Noortwijk, Jan M. van

    2009-01-01

    The paper presents a sampling-inspection strategy for the evaluation of time-dependent reliability of deteriorating systems, where the deterioration is assumed to initiate at random times and at random locations. After initiation, defects are weakening the system's resistance. The system becomes unacceptable when at least one defect reaches a critical depth. The defects are assumed to initiate at random times modeled as event times of a non-homogeneous Poisson process (NHPP) and to develop according to a non-decreasing time-dependent gamma process. The intensity rate of the NHPP is assumed to be a combination of a known time-dependent shape function and an unknown proportionality constant. When sampling inspection (i.e. inspection of a selected subregion of the system) results in a number of defect initiations, Bayes' theorem can be used to update prior beliefs about the proportionality constant of the NHPP intensity rate to the posterior distribution. On the basis of a time- and space-dependent Poisson process for the defect initiation, an adaptive Bayesian model for sampling inspection is developed to determine the predictive probability distribution of the time to failure. A potential application is, for instance, the inspection of a large vessel or pipeline suffering pitting/localized corrosion in the oil industry. The possibility of imperfect defect detection is also incorporated in the model.

  11. Simultaneous Voltammetric Detection of Carbaryl and Paraquat Pesticides on Graphene-Modified Boron-Doped Diamond Electrode

    NARCIS (Netherlands)

    Pop, Aniela; Manea, Florica; Flueras, Adriana; Schoonman, J.

    2017-01-01

    Monitoring of pesticide residues in food, beverages, and the environment requires fast, versatile, and sensitive analyzing methods. Direct electrochemical detection of pesticides could represent an efficient solution. Adequate electrode material, electrochemical technique, and optimal operation

  12. Simultaneous determination of ethyl carbamate and urea in Korean rice wine by ultra-performance liquid chromatography coupled with mass spectrometric detection.

    Science.gov (United States)

    Lee, Gyeong-Hweon; Bang, Dae-Young; Lim, Jung-Hoon; Yoon, Seok-Min; Yea, Myeong-Jai; Chi, Young-Min

    2017-10-15

    In this study, a rapid method for simultaneous detection of ethyl carbamate (EC) and urea in Korean rice wine was developed. To achieve quantitative analysis of EC and urea, the conditions for Ultra-performance liquid chromatography (UPLC) separation and atmospheric-pressure chemical ionization tandem mass spectrometry (APCI-MS/MS) detection were first optimized. Under the established conditions, the detection limit, relative standard deviation and linear range were 2.83μg/L, 3.75-5.96%, and 0.01-10.0mg/L, respectively, for urea; the corresponding values were 0.17μg/L, 1.06-4.01%, and 1.0-50.0μg/L, respectively, for EC. The correlation between the contents of EC and its precursor urea was determined under specific pH (3.5 and 4.5) and temperature (4, 25, and 50°C) conditions using the developed method. As a result, EC content was increased with greater temperature and lower pH. In Korean rice wine, urea was detected 0.19-1.37mg/L and EC was detected 2.0-7.7μg/L. The method developed in this study, which has the advantages of simplified sample preparation, low detection limits, and good selectivity, was successfully applied for the rapid analysis of EC and urea. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Two-Photon Probes for Lysosomes and Mitochondria: Simultaneous Detection of Lysosomes and Mitochondria in Live Tissues by Dual-Color Two-Photon Microscopy Imaging.

    Science.gov (United States)

    Lim, Chang Su; Hong, Seung Taek; Ryu, Seong Shick; Kang, Dong Eun; Cho, Bong Rae

    2015-10-01

    Novel two-photon (TP) probes were developed for lysosomes (PLT-yellow) and mitochondria (BMT-blue and PMT-yellow). These probes emitted strong TP-excited fluorescence in cells at widely separated wavelength regions and displayed high organelle selectivity, good cell permeability, low cytotoxicity, and pH insensitivity. The BMT-blue and PLT-yellow probes could be utilized to detect lysosomes and mitochondria simultaneously in live tissues by using dual-color two-photon microscopy, with minimum interference from each other. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Reliability, standard error, and minimum detectable change of clinical pressure pain threshold testing in people with and without acute neck pain.

    Science.gov (United States)

    Walton, David M; Macdermid, Joy C; Nielson, Warren; Teasell, Robert W; Chiasson, Marco; Brown, Lauren

    2011-09-01

    Clinical measurement. To evaluate the intrarater, interrater, and test-retest reliability of an accessible digital algometer, and to determine the minimum detectable change in normal healthy individuals and a clinical population with neck pain. Pressure pain threshold testing may be a valuable assessment and prognostic indicator for people with neck pain. To date, most of this research has been completed using algometers that are too resource intensive for routine clinical use. Novice raters (physiotherapy students or clinical physiotherapists) were trained to perform algometry testing over 2 clinically relevant sites: the angle of the upper trapezius and the belly of the tibialis anterior. A convenience sample of normal healthy individuals and a clinical sample of people with neck pain were tested by 2 different raters (all participants) and on 2 different days (healthy participants only). Intraclass correlation coefficient (ICC), standard error of measurement, and minimum detectable change were calculated. A total of 60 healthy volunteers and 40 people with neck pain were recruited. Intrarater reliability was almost perfect (ICC = 0.94-0.97), interrater reliability was substantial to near perfect (ICC = 0.79-0.90), and test-retest reliability was substantial (ICC = 0.76-0.79). Smaller change was detectable in the trapezius compared to the tibialis anterior. This study provides evidence that novice raters can perform digital algometry with adequate reliability for research and clinical use in people with and without neck pain.

  15. Detection of Alkynes via Click Chemistry with a Brominated Coumarin Azide by Simultaneous Fluorescence and Isotopic Signatures in Mass Spectrometry.

    Science.gov (United States)

    Yang, Lihua; Chumsae, Chris; Kaplan, Jenifer B; Moulton, Kevin Ryan; Wang, Dongdong; Lee, David H; Zhou, Zhaohui Sunny

    2017-09-20

    Alkynes are a key component of click chemistry and used for a wide variety of applications including bioconjugation, selective tagging of protein modifications, and labeling of metabolites and drug targets. However, challenges still exist for detecting alkynes because most 1,2,3-triazole products from alkynes and azides do not possess distinct intrinsic properties that can be used for their facile detection by either fluorescence or mass spectrometry. To address this critical need, a novel brominated coumarin azide was used to tag alkynes and detect alkyne-conjugated biomolecules. This tag has several useful properties: first, it is fluorogenic and the click-chemistry products are highly fluorescent and quantifiable; second, its distinct isotopic pattern facilitates identification by mass spectrometry; and third, its click-chemistry products form a unique pair of reporter ions upon fragmentation that can be used for the quick screening of data. Using a monoclonal antibody conjugated with alkynes, a general workflow has been developed and examined comprehensively.

  16. The reliability and accuracy of two methods for proximal caries detection and depth on directly visible proximal surfaces: an in vitro study

    DEFF Research Database (Denmark)

    Ekstrand, K R; Alloza, Alvaro Luna; Promisiero, L

    2011-01-01

    This study aimed to determine the reliability and accuracy of the ICDAS and radiographs in detecting and estimating the depth of proximal lesions on extracted teeth. The lesions were visible to the naked eye. Three trained examiners scored a total of 132 sound/carious proximal surfaces from 106 p...

  17. The current incidence of viral disease in korean sweet potatoes and development of multiplex rt-PCR assays for simultaneous detection of eight sweet potato viruses.

    Science.gov (United States)

    Kwak, Hae-Ryun; Kim, Mi-Kyeong; Shin, Jun-Chul; Lee, Ye-Ji; Seo, Jang-Kyun; Lee, Hyeong-Un; Jung, Mi-Nam; Kim, Sun-Hyung; Choi, Hong-Soo

    2014-12-01

    Sweet potato is grown extensively from tropical to temperate regions and is an important food crop worldwide. In this study, we established detection methods for 17 major sweet potato viruses using single and multiplex RT-PCR assays. To investigate the current incidence of viral diseases, we collected 154 samples of various sweet potato cultivars showing virus-like symptoms from 40 fields in 10 Korean regions, and analyzed them by RT-PCR using specific primers for each of the 17 viruses. Of the 17 possible viruses, we detected eight in our samples. Sweet potato feathery mottle virus (SPFMV) and sweet potato virus C (SPVC) were most commonly detected, infecting approximately 87% and 85% of samples, respectively. Furthermore, Sweet potato symptomless virus 1 (SPSMV-1), Sweet potato virus G (SPVG), Sweet potato leaf curl virus (SPLCV), Sweet potato virus 2 ( SPV2), Sweet potato chlorotic fleck virus (SPCFV), and Sweet potato latent virus (SPLV) were detected in 67%, 58%, 47%, 41%, 31%, and 20% of samples, respectively. This study presents the first documented occurrence of four viruses (SPVC, SPV2, SPCFV, and SPSMV-1) in Korea. Based on the results of our survey, we developed multiplex RT-PCR assays for simple and simultaneous detection of the eight sweet potato viruses we recorded.

  18. CMsearch: simultaneous exploration of protein sequence space and structure space improves not only protein homology detection but also protein structure prediction

    KAUST Repository

    Cui, Xuefeng

    2016-06-15

    Motivation: Protein homology detection, a fundamental problem in computational biology, is an indispensable step toward predicting protein structures and understanding protein functions. Despite the advances in recent decades on sequence alignment, threading and alignment-free methods, protein homology detection remains a challenging open problem. Recently, network methods that try to find transitive paths in the protein structure space demonstrate the importance of incorporating network information of the structure space. Yet, current methods merge the sequence space and the structure space into a single space, and thus introduce inconsistency in combining different sources of information. Method: We present a novel network-based protein homology detection method, CMsearch, based on cross-modal learning. Instead of exploring a single network built from the mixture of sequence and structure space information, CMsearch builds two separate networks to represent the sequence space and the structure space. It then learns sequence–structure correlation by simultaneously taking sequence information, structure information, sequence space information and structure space information into consideration. Results: We tested CMsearch on two challenging tasks, protein homology detection and protein structure prediction, by querying all 8332 PDB40 proteins. Our results demonstrate that CMsearch is insensitive to the similarity metrics used to define the sequence and the structure spaces. By using HMM–HMM alignment as the sequence similarity metric, CMsearch clearly outperforms state-of-the-art homology detection methods and the CASP-winning template-based protein structure prediction methods.

  19. The Current Incidence of Viral Disease in Korean Sweet Potatoes and Development of Multiplex RT-PCR Assays for Simultaneous Detection of Eight Sweet Potato Viruses

    Directory of Open Access Journals (Sweden)

    Hae-Ryun Kwak

    2014-12-01

    Full Text Available Sweet potato is grown extensively from tropical to temperate regions and is an important food crop worldwide. In this study, we established detection methods for 17 major sweet potato viruses using single and multiplex RT-PCR assays. To investigate the current incidence of viral diseases, we collected 154 samples of various sweet potato cultivars showing virus-like symptoms from 40 fields in 10 Korean regions, and analyzed them by RT-PCR using specific primers for each of the 17 viruses. Of the 17 possible viruses, we detected eight in our samples. Sweet potato feathery mottle virus (SPFMV and sweet potato virus C (SPVC were most commonly detected, infecting approximately 87% and 85% of samples, respectively. Furthermore, Sweet potato symptomless virus 1 (SPSMV-1, Sweet potato virus G (SPVG, Sweet potato leaf curl virus (SPLCV, Sweet potato virus 2 ( SPV2, Sweet potato chlorotic fleck virus (SPCFV, and Sweet potato latent virus (SPLV were detected in 67%, 58%, 47%, 41%, 31%, and 20% of samples, respectively. This study presents the first documented occurrence of four viruses (SPVC, SPV2, SPCFV, and SPSMV-1 in Korea. Based on the results of our survey, we developed multiplex RT-PCR assays for simple and simultaneous detection of the eight sweet potato viruses we recorded.

  20. A technique for simultaneous detection of individual vortex states of Laguerre-Gaussian beams transmitted through an aqueous suspension of microparticles

    Science.gov (United States)

    Khonina, S. N.; Karpeev, S. V.; Paranin, V. D.

    2018-06-01

    A technique for simultaneous detection of individual vortex states of the beams propagating in a randomly inhomogeneous medium is proposed. The developed optical system relies on the correlation method that is invariant to the beam wandering. The intensity distribution formed at the optical system output does not require digital processing. The proposed technique based on a multi-order phase diffractive optical element (DOE) is studied numerically and experimentally. The developed detection technique is used for the analysis of Laguerre-Gaussian vortex beams propagating under conditions of intense absorption, reflection, and scattering in transparent and opaque microparticles in aqueous suspensions. The performed experimental studies confirm the relevance of the vortex phase dependence of a laser beam under conditions of significant absorption, reflection, and scattering of the light.

  1. Simultaneous detection and serotyping of Salmonellae by immunomagnetic separation and label-free surface enhanced Raman spectroscopy

    Science.gov (United States)

    Salmonella spp. are one of the leading causes of foodborne outbreaks in the United States and globally. Current detection and characterization techniques for Salmonellae are time consuming and costly, and rapid methods could greatly benefit outbreak investigation, new case prevention and disease tre...

  2. Development of a nucleic acid lateral flow immunoassay for simultaneous detection of Listeria spp. and Listeriamonocytogenes in food

    NARCIS (Netherlands)

    Blazkova, M.; Koets, M.; Rauch, P.; Amerongen, van A.

    2009-01-01

    We present a new nucleic acid lateral flow immunoassay (NALFIA) for the assessment of listeria contamination. The detection procedure starts with enrichment of sample in Half Fraser broth (24 h). Following isolation of DNA, a duplex PCR is performed with two labelled primer sets, one generic and

  3. Strip-based immunoassay for the simultaneous detection of the neonicotinoid insecticides imidacloprid and thiamethoxam in agricultural products

    Science.gov (United States)

    A semiquantitative strip immunoassay was developed for the rapid detection of imidacloprid and thiamethoxam in agricultural products using specific nanocolloidal gold-labeled monoclonal antibodies. The conjugates of imidacloprid-BSA and thiamethoxam-BSA and goat anti-mouse IgG were coated on the ni...

  4. The Cu-MOF-199/single-walled carbon nanotubes modified electrode for simultaneous determination of hydroquinone and catechol with extended linear ranges and lower detection limits

    International Nuclear Information System (INIS)

    Zhou, Jian; Li, Xi; Yang, Linlin; Yan, Songlin; Wang, Mengmeng; Cheng, Dan; Chen, Qi; Dong, Yulin; Liu, Peng; Cai, Weiquan; Zhang, Chaocan

    2015-01-01

    A novel electrochemical sensor based on Cu-MOF-199 [Cu-MOF-199 = Cu 3 (BTC) 2 (BTC = 1,3,5-benzenetricarboxylicacid)] and SWCNTs (single-walled carbon nanotubes) was fabricated for the simultaneous determination of hydroquinone (HQ) and catechol (CT). The modification procedure was carried out through casting SWCNTs on the bare glassy carbon electrode (GCE) and followed by the electrodeposition of Cu-MOF-199 on the SWCNTs modified electrode. Cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM) were performed to characterize the electrochemical performance and surface characteristics of the as-prepared sensor. The composite electrode exhibited an excellent electrocatalytic activity with increased electrochemical signals towards the oxidation of HQ and CT, owing to the synergistic effect of SWCNTs and Cu-MOF-199. Under the optimized condition, the linear response range were from 0.1 to 1453 μmol L −1 (R HQ  = 0.9999) for HQ and 0.1–1150 μmol L −1 (R CT  = 0.9990) for CT. The detection limits for HQ and CT were as low as 0.08 and 0.1 μmol L −1 , respectively. Moreover, the modified electrode presented the good reproducibility and the excellent anti-interference performance. The analytical performance of the developed sensor for the simultaneous detection of HQ and CT had been evaluated in practical samples with satisfying results. - Highlights: • Cu-MOF-199/SWCNTs/GCE was facilely fabricated by the electrodeposition on SWCNTs/GCE. • An electrochemical sensor for detecting HQ and CT was constructed based on this modified electrode. • The proposed electrochemical sensor showed an extended linear range and lower detection limits. • The proposed electrochemical sensor had an excellent stability and reproducibility.

  5. Multiplex real-time PCR (TaqMan) assay for the simultaneous detection and discrimination of potato powdery and common scab diseases and pathogens.

    Science.gov (United States)

    Qu, X S; Wanner, L A; Christ, B J

    2011-03-01

    To develop a multiplex real-time PCR assay using TaqMan probes for the simultaneous detection and discrimination of potato powdery scab and common scab, two potato tuber diseases with similar symptoms, and the causal pathogens Spongospora subterranea and plant pathogenic Streptomyces spp. Real-time PCR primers and a probe for S. subterranea were designed based on the DNA sequence of the ribosomal RNA ITS2 region. Primers and a probe for pathogenic Streptomyces were designed based on the DNA sequence of the txtAB genes. The two sets of primer pairs and probes were used in a single real-time PCR assay. The multiplex real-time PCR assay was confirmed to be specific for S. subterranea and pathogenic Streptomyces. The assay detected DNA quantities of 100 fg for each of the two pathogens and linear responses and high correlation coefficients between the amount of DNA and C(t) values for each pathogen were achieved. The presence of two sets of primer pairs and probes and of plant extracts did not alter the sensitivity and efficiency of multiplex PCR amplification. Using the PCR assay, we could discriminate between powdery scab and common scab tubers with similar symptoms. Common scab and powdery scab were detected in some tubers with no visible symptoms. Mixed infections of common scab and powdery scab on single tubers were also revealed. This multiplex real-time PCR assay is a rapid, cost efficient, specific and sensitive tool for the simultaneous detection and discrimination of the two pathogens on infected potato tubers when visual symptoms are inconclusive or not present. Accurate and quick identification and discrimination of the cause of scab diseases on potatoes will provide critical information to potato growers and researchers for disease management. This is important because management strategies for common and powdery scab diseases are very different. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  6. Improvement of Matrix Converter Drive Reliability by Online Fault Detection and a Fault-Tolerant Switching Strategy

    DEFF Research Database (Denmark)

    Nguyen-Duy, Khiem; Liu, Tian-Hua; Chen, Der-Fa

    2011-01-01

    The matrix converter system is becoming a very promising candidate to replace the conventional two-stage ac/dc/ac converter, but system reliability remains an open issue. The most common reliability problem is that a bidirectional switch has an open-switch fault during operation. In this paper, a...

  7. Simultaneous detection of ascorbic acid, dopamine, uric acid and tryptophan with Azure A-interlinked multi-walled carbon nanotube/gold nanoparticles composite modified electrode

    Directory of Open Access Journals (Sweden)

    Hayati Filik

    2016-05-01

    Full Text Available In this paper, multi-walled carbon nanotube/Azure A/gold nanoparticle composites (Nafion/AuNPs/AzA/MWCNTs were prepared by binding gold nanoparticles to the surfaces of Azure A-coated carbon nanotubes. Nafion/AuNPs/AzA/MWCNTs based electrochemical sensor was fabricated for the simultaneous determination of ascorbic acid, dopamine, uric acid, and tryptophan. Cyclic voltammetry and electrochemical impedance spectroscopy were used to characterize the electrochemical properties of the modified electrodes. The modified electrode showed excellent electrocatalytic activity toward ascorbic acid, dopamine, uric acid, and tryptophan (pH 7.0. The experiment results showed that the linear response range for simultaneous detection of AA, DA, UA and Trp were 300–10,000 μM, 0.5–50 μM, 0.5–50 μM and 1.0–100 μM, respectively, and the detection limits were 16 μM, 0.014 μM, 0.028 μM and 0.56 μM (S/N = 3. The proposed method offers promise for simple, rapid, selective and cost-effective analysis of small biomolecules. The procedure was also applied to the determination of tryptophan in spiked milk samples.

  8. A sensitive HPLC-MS/MS method for the simultaneous detection of microbial transglutaminase, and bovine and porcine fibrinogen/thrombin in restructured meat.

    Science.gov (United States)

    Jira, Wolfgang; Schwägele, Fredi

    2017-12-15

    A sensitive HPLC-MS/MS method for the simultaneous detection of microbial transglutaminase (TG) from Streptomyces mobaraensis, and bovine and porcine fibrinogen/thrombin in restructured meat was developed using tryptic marker peptides of TG (five markers), and bovine and porcine fibrinogen (six markers each). Meat binding experiments with beef and pork were performed using a technical TG mixture (Activa, Ajinomoto), and bovine and porcine plasmapowder FG (PPFG; Sonac B.V.). The method developed allows the simultaneous detection of the use of these cold-set binders in raw and heated samples. The peak areas of the fibrinogen marker peptides were increased by a factor of about 100, compared to blank values originating from the occurrence of residual blood in meat, using a concentration of 0.6% bovine and porcine PPFG. A differentiation between the use of blood plasma powder and PPFG using the ratios of fibrinogen to serotransferrin peptide peak areas seems to be possible. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Ascorbic acid surface modified TiO₂-thin layers as a fully integrated analysis system for visual simultaneous detection of organophosphorus pesticides.

    Science.gov (United States)

    Li, Shunxing; Liang, Wenjie; Zheng, Fengying; Lin, Xiaofeng; Cai, Jiabai

    2014-11-06

    TiO₂ photocatalysis and colorimetric detection are coupled with thin layer chromatography (TLC) for the first time to develop a fully integrated analysis system. Titania@polystyrene hybrid microspheres were surface modified with ascorbic acid, denoted AA-TiO₂@PS, and used as the stationary phase for TLC. Because the affinity between AA-TiO₂@PS and organophosphorus pesticides (OPs) was different for different species of OPs (including chlopyrifos, malathion, parathion, parathion-methyl, and methamidophos), OPs could be separated simultaneously by the mobile phase in 12.0 min with different Rf values. After surface modification, the UV-vis wavelength response range of AA-TiO₂@PS was expanded to 650 nm. Under visible-light irradiation, all of the OPs could be photodegraded to PO₄(3-) in 25.0 min. Based on the chromogenic reaction between PO₄(3-) and chromogenic agents (ammonium molybdate and ascorbic acid), OPs were quantified from color intensity images using a scanner in conjunction with image processing software. So, AA-TiO₂@PS was respectively used as the stationary phase of TLC for efficient separation of OPs, as a photocatalyst for species transformation of phosphorus, and as a colorimetric probe for on-field simultaneous visual detection of OPs in natural water. Linear calibration curves for each OP ranged from 19.3 nmol P L(-1) to 2.30 μmol P L(-1). This integrated analysis system was simple, inexpensive, easy to operate, and sensitive.

  10. Simultaneous determination of flavonoids, isochlorogenic acids and triterpenoids in Ilex hainanensis Using high performance liquid chromatography coupled with diode array and evaporative light scattering detection.

    Science.gov (United States)

    Peng, Bo; Qiao, Chun-Feng; Zhao, Jing; Huang, Wei-Hua; Hu, De-Jun; Liu, Hua-Gang; Li, Shao-Ping

    2013-03-04

    A high performance liquid chromatography coupled with diode array and evaporative light scattering detection (HPLC-DAD-ELSD) method for simultaneous determination of eight major bioactive compounds including two flavonoids (rutin and eriodictyol-7-O-β-D-glucopyranoside), two isochlorogenic acids (isochlorogenic acid A and isochlorogenic acid C) and four triterpenoids (ilexhainanoside D, ilexsaponin A1, ilexgenin A and ursolic acid) in Ilex hainanensis has been developed for the first time. The 283 nm wavelength was chosen for determination of two flavonoids and two isochlorogenic acids. ELSD was applied to determine four triterpenoids. The analysis was performed on an Agilent Zorbax SB-C18 column (250 × 4.6 mm i.d., 5 µm) with gradient elution of 0.2% formic acid in water and acetonitrile. The method was validated for linearity, limit of detection, limit of quantification, precision, repeatability and accuracy. The proposed method has been successfully applied for simultaneous quantification of the analytes in four samples of Ilex hainanensis, which is helpful for quality control of this plant.

  11. Metabolic cytometry: capillary electrophoresis with two-color fluorescence detection for the simultaneous study of two glycosphingolipid metabolic pathways in single primary neurons.

    Science.gov (United States)

    Essaka, David C; Prendergast, Jillian; Keithley, Richard B; Palcic, Monica M; Hindsgaul, Ole; Schnaar, Ronald L; Dovichi, Norman J

    2012-03-20

    Metabolic cytometry is a form of chemical cytometry wherein metabolic cascades are monitored in single cells. We report the first example of metabolic cytometry where two different metabolic pathways are simultaneously monitored. Glycolipid catabolism in primary rat cerebella neurons was probed by incubation with tetramethylrhodamine-labeled GM1 (GM1-TMR). Simultaneously, both catabolism and anabolism were probed by coincubation with BODIPY-FL labeled LacCer (LacCer-BODIPY-FL). In a metabolic cytometry experiment, single cells were incubated with substrate, washed, aspirated into a capillary, and lysed. The components were separated by capillary electrophoresis equipped with a two-spectral channel laser-induced fluorescence detector. One channel monitored fluorescence generated by the metabolic products produced from GM1-TMR and the other monitored the metabolic products produced from LacCer-BODIPY-FL. The metabolic products were identified by comparison with the mobility of a set of standards. The detection system produced at least 6 orders of magnitude dynamic range in each spectral channel with negligible spectral crosstalk. Detection limits were 1 zmol for BODIPY-FL and 500 ymol for tetramethylrhodamine standard solutions.

  12. Reliability and minimal detectable change of a modified passive neck flexion test in patients with chronic nonspecific neck pain and asymptomatic subjects.

    Science.gov (United States)

    López-de-Uralde-Villanueva, Ibai; Acuyo-Osorio, Mario; Prieto-Aldana, María; La Touche, Roy

    2017-04-01

    The Passive Neck Flexion Test (PNFT) can diagnose meningitis and potential spinal disorders. Little evidence is available concerning the use of a modified version of the PNFT (mPNFT) in patients with chronic nonspecific neck pain (CNSNP). To assess the reliability of the mPNFT in subjects with and without CNSNP. The secondary objective was to assess the differences in the symptoms provoked by the mPNFT between these two populations. We used repeated measures concordance design for the main objective and cross-sectional design for the secondary objective. A total of 30 asymptomatic subjects and 34 patients with CNSNP were recruited. The following measures were recorded: the range of motion at the onset of symptoms (OS-mPNFT), the range of motion at the submaximal pain (SP-mPNFT), and evoked pain intensity on the mPNFT (VAS-mPNFT). Good to excellent reliability was observed for OS-mPNFT and SP-mPNFT in the asymptomatic group (intra-examiner reliability: 0.95-0.97; inter-examiner reliability: 0.86-0.90; intra-examiner test-retest reliability: 0.84-0.87). In the CNSNP group, a good to excellent reliability was obtained for the OS-mPNFT (intra-examiner reliability: 0.89-0.96; inter-examiner reliability: 0.83-0.86; intra-examiner test-retest reliability: 0.83-0.85) and the SP-PNFT (intra-examiner reliability: 0.94-0.98; inter-examiner reliability: 0.80-0.82; intra-examiner test-retest reliability: 0.88-0.91). The CNSNP group showed statistically significant differences in OS-mPNFT (t = 4.92; P reliable tool regardless of the examiner and the time factor. Patients with CNSNP have a decrease range of motion and more pain than asymptomatic subjects in the mPNFT. This exceeds the minimal detectable changes for OS-mPNFT and VAS-mPNFT. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Simultaneous application of multiple platforms (Glider, Scanfish, profiling mooring, CTD) to improve detection and quantification of temporal ocean dynamics

    Science.gov (United States)

    Meyer, D.; Prien, R. D.; Lips, U.; Naumann, M.; Liblik, T.; Schulz-Bull, D. E.

    2016-02-01

    Ocean dynamics are difficult to observe given the broad spectrum of temporal and spatial scales. Robotic technology can be used to address this issue, and help to investigate the variability of physical and biogeochemical processes. This work focuses on ocean robots and in particular on glider technology which seems to be one of the most promising oceanographic tools for future marine research. In this context, we present the results of an observational program conducted in the Baltic Sea combining a profiling mooring (GODESS - Gotland Deep Environmental Sampling Station) and glider technology (Slocum). The temporal variability is captured by the mooring, while the spatial variability is obtained from the glider sampling the surrounding area. Furthermore, classical CTD-measurements and an underwater vehicle (Scanfish) are used simultaneously by two different research vessels to validate and complement the observing network. The main aim of the study is to identify possible synergies between the different platforms and to get a better understanding of maximizing the information content of the data collected by this network. The value and the quality of the data of each individual platform is analyzed and their contribution to the performance of the network itself evaluated.

  14. Direct and simultaneous detection of organic and inorganic ingredients in herbal powder preparations by Fourier transform infrared microspectroscopic imaging.

    Science.gov (United States)

    Chen, Jian-Bo; Sun, Su-Qin; Tang, Xu-Dong; Zhang, Jing-Zhao; Zhou, Qun

    2016-08-05

    Herbal powder preparation is a kind of widely-used herbal product in the form of powder mixture of herbal ingredients. Identification of herbal ingredients is the first and foremost step in assuring the quality, safety and efficacy of herbal powder preparations. In this research, Fourier transform infrared (FT-IR) microspectroscopic identification method is proposed for the direct and simultaneous recognition of multiple organic and inorganic ingredients in herbal powder preparations. First, the reference spectrum of characteristic particles of each herbal ingredient is assigned according to FT-IR results and other available information. Next, a statistical correlation threshold is determined as the lower limit of correlation coefficients between the reference spectrum and a larger number of calibration characteristic particles. After validation, the reference spectrum and correlation threshold can be used to identify herbal ingredient in mixture preparations. A herbal ingredient is supposed to be present if correlation coefficients between the reference spectrum and some sample particles are above the threshold. Using this method, all kinds of herbal materials in powder preparation Kouqiang Kuiyang San are identified successfully. This research shows the potential of FT-IR microspectroscopic identification method for the accurate and quick identification of ingredients in herbal powder preparations. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Simultaneous detection of hepatitis B virus genotypes and mutations associated with resistance to lamivudine, adefovir, and telbivudine by the polymerase chain reaction-ligase detection reaction

    Directory of Open Access Journals (Sweden)

    Yong-Zhong Wang

    Full Text Available OBJECTIVES: Detection of mutations associated to nucleos(tide analogs and hepatitis B virus (HBV genotyping are essential for monitoring treatment of HBV infection. We developed a multiplex polymerase chain reaction-ligase detection reaction (PCR-LDR assay for the rapid detection of HBV genotypes and mutations associated with lamivudine, adefovir, and telbivudine resistance in HBV-infected patients. METHODS: HBV templates were amplified by PCR, followed by LDR and electrophoresis on a sequencer. The assay was evaluated using plasmids that contained wild-type or mutant HBV sequences and 216 clinical samples. RESULTS: The PCR-LDR assay and sequencing gave comparable results for 158 of the 216 samples (73.1% with respect to mutation detection and genotyping. Complete agreement between the two methods was observed for all the samples (100% at codon 180 and codon 204. Concordant results were observed for 99.4% of the 158 samples at codon 181 and 98.7% at codon 236. The genotyping results were completely concordant between the PCR-LDR assay and sequencing. The PCR-LDR assay could detect a proportion of 1% mutant plasmid in a background of wild-type plasmid. CONCLUSION: The PCR-LDR assay is sensitive and specific for detection of HBV genotypes and drug resistance mutations, and could be helpful for decision making in the treatment of HBV infection.

  16. Simultaneous determination of active ingredients in Erigeron breviscapus (Vant.) Hand-Mazz. by capillary electrophoresis with electrochemical detection.

    Science.gov (United States)

    Chu, Qingcui; Wu, Ting; Fu, Liang; Ye, Jiannong

    2005-03-09

    A high-performance capillary electrophoresis (CE) with electrochemical detection (ED) method was developed for the determination of the pharmacologically active ingredients in Erigeron breviscapus (Vant.) Hand-Mazz. and its extract phytopharmaceuticals in this work. Under the optimum conditions, nine analytes, baicalein, naringenin, scopoletin, kaempferol, apigenin, scutellarin, luteolin, caffeic acid and protocatechuic acid were separated within 24 min in a borax buffer (pH 8.7). Notably, excellent linearity was obtained over two orders of magnitude with detection limits (S/N=3) ranged from 1.0 x 10(-7) g/mL to 5.6 x 10(-7) g/mL for all nine analytes. This method was successfully used in the analysis of E. breviscapus (Vant.) Hand-Mazz. and its phytopharmaceuticals with a relatively simple extraction procedure, and the assay results were satisfactory.

  17. Simultaneous detection of enteropathogenic viruses in buffalos faeces using multiplex reverse transcription-polymerase chain reaction (mRT-PCR

    Directory of Open Access Journals (Sweden)

    U. Pagnini

    2010-02-01

    Full Text Available A multiplex reverse transcription- polymerase chain reaction (mRT-PCR assay that detects Bovine Viral Diarrhoea Virus, Bovine Coronavirus, and Group A Rotaviruses in infected cell-culture fluids and clinical faecal samples is described. One hundred twenty faecal samples from buffalo calves with acute gastroenteritis were tested. The mRT-PCR was validated against simplex RT-PCR with published primers for Pestivirus, Coronavirus and Rotavirus. The multiplex RT-PCR was equally sensitive and specific in detecting viral infections compared with simplex RT-PCR. The mRT-PCR readily identified viruses by discriminating the size of their amplified gene products. This mRT-PCR may be a sensitive and rapid assay for surveillance of buffalo enteric viruses in field specimens. This novel multiplex RT-PCR is an attractive technique for the rapid, specific, and cost-effective laboratory diagnosis of acute gastroenteritis.

  18. A simple, one-tube assay for the simultaneous detection and diagnosis of ten Australian poultry Eimeria.

    Science.gov (United States)

    Godwin, Rosamond M; Morgan, Jess A T

    2014-02-01

    Coccidiosis is a costly worldwide enteric disease of chickens caused by parasites of the genus Eimeria. At present, there are seven described species that occur globally and a further three undescribed, operational taxonomic units (OTUs X, Y, and Z) that are known to infect chickens from Australia. Species of Eimeria have both overlapping morphology and pathology and frequently occur as mixed-species infections. This makes definitive diagnosis with currently available tests difficult and, to date, there is no test for the detection of the three OTUs. This paper describes the development of a PCR-based assay that is capable of detecting all ten species of Eimeria, including OTUs X, Y, and Z in field samples. The assay is based on a single set of generic primers that amplifies a single diagnostic fragment from the mitochondrial genome of each species. This one-tube assay is simple, low-cost, and has the capacity to be high throughput. It will therefore be of great benefit to the poultry industry for Eimeria detection and control, and the confirmation of identity and purity of vaccine strains. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Development of a real-time PCR melt curve assay for simultaneous detection of virulent and antibiotic resistant Salmonella.

    Science.gov (United States)

    Singh, Prashant; Mustapha, Azlin

    2014-12-01

    Multiple drug resistance in Salmonella is an emerging problem in the area of food safety. Depending on the virulence and antibiotic resistance characteristics of the Salmonella strain, infections of varying severity could result. In this study, a multiplex melt curve real-time PCR assay for the detection of virulent and antibiotic resistance strains of Salmonella was developed with two primer sets. The first set targets the virulence gene, invasin (invA), and tetracycline (tetG), streptomycin (aadA2) and sulphonamide (sulI) antibiotic resistance genes, and the second set amplifies ampicillin (blaPSE,blaTEM) and chloramphenicol (floR) resistance genes. The multiplex assay was evaluated using 41 Salmonella strains and was further tested on eight different artificially inoculated food samples. The fluorescent DNA intercalating dye, SYTO9, generated high resolution melt curve peaks and, hence, was used for the development of the assay. This multiplex assay worked efficiently over a DNA concentration range of 20 ng-200 fg and showed a sensitivity of 290 CFU/mL with serially diluted broth cultures. The detection limit for un-enriched artificially inoculated food samples was 10(4) CFU/g, but an enrichment period of 6 h allowed for detection of 10 CFU/g of cells in the samples. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. A fast, reliable, ultra high performance liquid chromatography method for the simultaneous determination of amino acids, biogenic amines and ammonium ions in cheese, using diethyl ethoxymethylenemalonate as a derivatising agent.

    Science.gov (United States)

    Redruello, Begoña; Ladero, Victor; Cuesta, Isabel; Álvarez-Buylla, Jorge R; Martín, María Cruz; Fernández, María; Alvarez, Miguel A

    2013-08-15

    Derivatisation treatment with diethyl ethoxymethylenemalonate followed by ultra-HPLC allowed the simultaneous quantification of 22 amino acids, 7 biogenic amines and ammonium ions in cheese samples in under 10 min. This is the fastest elution time ever reported for such a resolution. The proposed method shows good linearity (R(2)>0.995) and sensitivity (detection limit 0.08-3.91 μM; quantification limit <13.02 μM). Intra- and inter-day repeatability ranged from 0.35% to 1.25% and from 0.85% to 5.2%, respectively. No significant effect of the cheese matrix was observed. Copyright © 2013 Elsevier Ltd. All rights reserved.

  1. ZIF-67 derived porous Co3O4 hollow nanopolyhedron functionalized solution-gated graphene transistors for simultaneous detection of glucose and uric acid in tears.

    Science.gov (United States)

    Xiong, Can; Zhang, Tengfei; Kong, Weiyu; Zhang, Zhixiang; Qu, Hao; Chen, Wei; Wang, Yanbo; Luo, Linbao; Zheng, Lei

    2018-03-15

    Biomarkers in tears have attracted much attention in daily healthcare sensing and monitoring. Here, highly sensitive sensors for simultaneous detection of glucose and uric acid are successfully constructed based on solution-gated graphene transistors (SGGTs) with two separate Au gate electrodes, modified with GOx-CHIT and BSA-CHIT respectively. The sensitivity of the SGGT is dramatically improved by co-modifying the Au gate with ZIF-67 derived porous Co 3 O 4 hollow nanopolyhedrons. The sensing mechanism for glucose sensor is attributed to the reaction of H 2 O 2 generated by the oxidation of glucose near the gate, while the sensing mechanism for uric acid is due to the direct electro-oxidation of uric acid molecules on the gate. The optimized glucose and uric acid sensors show the detection limits both down to 100nM, far beyond the sensitivity required for non-invasive detection of glucose and uric acid in tears. The glucose and uric acid in real tear samples was quantitatively detected at 323.2 ± 16.1μM and 98.5 ± 16.3μM by using the functionalized SGGT device. Due to the low-cost, high-biocompatibility and easy-fabrication features of the ZIF-67 derived porous Co 3 O 4 hollow nanopolyhedron, they provide excellent electrocatalytic nanomaterials for enhancing sensitivity of SGGTs for a broad range of disease-related biomarkers. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Fast and simultaneous detection of prominent natural antioxidants using analytical microsystems for capillary electrophoresis with a glassy carbon electrode: a new gateway to food environments.

    Science.gov (United States)

    Blasco, Antonio Javier; Barrigas, Inés; González, María Cristina; Escarpa, Alberto

    2005-12-01

    This paper examines for the first time the analytical possibilities of fast and simultaneous detection of prominent natural antioxidants including examples of flavonoids and vitamins using a CE microchip with electrochemical detection (ED). Unpinched injection conditions, zone electrophoretic separation and amperometric detection were carefully assayed and optimised. Analysis involved the zone electrophoretic separation of arbutin, (+)-catechin and ascorbic acid in less than 4 min using a borate buffer (pH 9.0, 50 mM), employing 2 kV as the separation voltage and +1.0 V as the detection potential. In addition, the separation of different 'couples' of natural antioxidants of food significance including (+)-catechin and ascorbic acid, (+)-catechin and rutin, as well as arbutin and phlorizdin is proposed. To demonstrate the potential and future role of CE microsystems, analytical possibilities and a new route in the raw sample analysis are presented. The preliminary results obtained allow the proposal of CE-ED microchips as a real gateway to microanalysis in foods.

  3. Development and validation of a multiplex real-time PCR method to simultaneously detect 47 targets for the identification of genetically modified organisms.

    Science.gov (United States)

    Cottenet, Geoffrey; Blancpain, Carine; Sonnard, Véronique; Chuah, Poh Fong

    2013-08-01

    Considering the increase of the total cultivated land area dedicated to genetically modified organisms (GMO), the consumers' perception toward GMO and the need to comply with various local GMO legislations, efficient and accurate analytical methods are needed for their detection and identification. Considered as the gold standard for GMO analysis, the real-time polymerase chain reaction (RTi-PCR) technology was optimised to produce a high-throughput GMO screening method. Based on simultaneous 24 multiplex RTi-PCR running on a ready-to-use 384-well plate, this new procedure allows the detection and identification of 47 targets on seven samples in duplicate. To comply with GMO analytical quality requirements, a negative and a positive control were analysed in parallel. In addition, an internal positive control was also included in each reaction well for the detection of potential PCR inhibition. Tested on non-GM materials, on different GM events and on proficiency test samples, the method offered high specificity and sensitivity with an absolute limit of detection between 1 and 16 copies depending on the target. Easy to use, fast and cost efficient, this multiplex approach fits the purpose of GMO testing laboratories.

  4. Simultaneous detection of Ponceat 4R and tartrazine in food using adsorptive stripping voltammetry on an acetylene black nanoparticle-modified electrode.

    Science.gov (United States)

    Yang, Xiaofeng; Qin, Haibin; Gao, Miaomiao; Zhang, Huajie

    2011-12-01

    Ponceau 4R and tartrazine have been widely used in foodstuffs. However, they are pathogenic if they are excessively consumed. Therefore, the detection of Ponceat 4R and tartrazine is quite important. A sensitive and rapid electrochemical method was developed for the simultaneous detection of Ponceat 4R and tartrazine using anodic adsorptive stripping voltammetry and based on the strong enhancement effect of acetylene black nanoparticle. For Ponceat 4R, the linear range was from 0.05 to 4 mg kg(-1) , and the limit of detection was 0.03 mg kg(-1) . For tartrazine, the linear range was from 0.15 to 18 mg kg(-1) , and the limit of detection was 0.1 mg kg(-1) . The relative standard deviation was 3.8% and 4.7% for 10 successive measurements of 1 mg kg(-1) Ponceau 4R and tartrazine. The method was used to determine Ponceat 4R and tartrazine in soft drinks, and recovery was in the range of 92.4-104.8%. At the acetylene black nanoparticle-modified electrode, the oxidation current signal of Ponceau 4R and tartrazine greatly increase. This new method is sensitive, rapid, simple and feasible. Copyright © 2011 Society of Chemical Industry.

  5. Direct duplex real-time loop mediated isothermal amplification assay for the simultaneous detection of cow and goat species origin of milk and yogurt products for field use.

    Science.gov (United States)

    Kim, Mi-Ju; Kim, Hae-Yeong

    2018-04-25

    A multiple loop-mediated isothermal amplification (LAMP) method was developed to detect cow and goat milk in the field using a portable fluorescence device. For rapid on-site detection, this duplex LAMP assay was used in combination with direct amplification, without DNA extraction. The cow- and goat-specific LAMP primer sets were designed based on the mitochondrial cytochrome b gene, and showed specificity against 13 other animal species in the reactions. The sensitivity of the duplex LAMP assay for cow and goat was 0.1 and 1 pg, respectively. The detection limit for both target species in milk mixtures was 2%. This assay successfully amplified and identified the two target species in 24 samples of commercial milk and yogurt products, with 30 min sampling-to-result analysis time. Therefore, this direct duplex real-time LAMP assay is useful for on-site simultaneous detection of cow and goat milk in commercial products, a capability needed to confirm accurate labeling. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Simultaneous membrane interaction of amphipathic peptide monomers, self-aggregates and cargo complexes detected by fluorescence correlation spectroscopy.

    Science.gov (United States)

    Vasconcelos, Luís; Lehto, Tõnis; Madani, Fatemeh; Radoi, Vlad; Hällbrink, Mattias; Vukojević, Vladana; Langel, Ülo

    2018-02-01

    Peptides able to translocate cell membranes while carrying macromolecular cargo, as cell-penetrating peptides (CPPs), can contribute to the field of drug delivery by enabling the transport of otherwise membrane impermeable molecules. Formation of non-covalent complexes between amphipathic peptides and oligonucleotides is driven by electrostatic and hydrophobic interactions. Here we investigate and quantify the coexistence of distinct molecular species in multiple equilibria, namely peptide monomer, peptide self-aggregates and peptide/oligonucleotide complexes. As a model for the complexes, we used a stearylated peptide from the PepFect family, PF14 and siRNA. PF14 has a cationic part and a lipid part, resembling some characteristics of cationic lipids. Fluorescence correlation spectroscopy (FCS) and fluorescence cross-correlation spectroscopy (FCCS) were used to detect distinct molecular entities in solution and at the plasma membrane of live cells. For that, we labeled the peptide with carboxyrhodamine 6G and the siRNA with Cyanine 5. We were able to detect fluorescent entities with diffusional properties characteristic of the peptide monomer as well as of peptide aggregates and peptide/oligonucleotide complexes. Strategies to avoid peptide adsorption to solid surfaces and self-aggregation were developed and allowed successful FCS measurements in solution and at the plasma membrane. The ratio between the detected molecular species was found to vary with pH, peptide concentration and the proximity to the plasma membrane. The present results suggest that the diverse cellular uptake mechanisms, often reported for amphipathic CPPs, might result from the synergistic effect of peptide monomers, self-aggregates and cargo complexes, distributed unevenly at the plasma membrane. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Simultaneous detection of multiple HPV DNA via bottom-well microfluidic chip within an infra-red PCR platform.

    Science.gov (United States)

    Liu, Wenjia; Warden, Antony; Sun, Jiahui; Shen, Guangxia; Ding, Xianting

    2018-03-01

    Portable Polymerase Chain Reaction (PCR) devices combined with microfluidic chips or lateral flow stripes have shown great potential in the field of point-of-need testing (PoNT) as they only require a small volume of patient sample and are capable of presenting results in a short time. However, the detection for multiple targets in this field leaves much to be desired. Herein, we introduce a novel PCR platform by integrating a bottom-well microfluidic chip with an infra-red (IR) excited temperature control method and fluorescence co-detection of three PCR products. Microfluidic chips are utilized to partition different samples into individual bottom-wells. The oil phase in the main channel contains multi-walled carbon nanotubes which were used as a heat transfer medium that absorbs energy from the IR-light-emitting diode (LED) and transfers heat to the water phase below. Cyclical rapid heating and cooling necessary for PCR are achieved by alternative power switching of the IR-LED and Universal Serial Bus (USB) mini-fan with a pulse width modulation scheme. This design of the IR-LED PCR platform is economic, compact, and fully portable, making it a promising application in the field of PoNT. The bottom-well microfluidic chip and IR-LED PCR platform were combined to fulfill a three-stage thermal cycling PCR for 40 cycles within 90 min for Human Papilloma Virus (HPV) detection. The PCR fluorescent signal was successfully captured at the end of each cycle. The technique introduced here has broad applications in nucleic acid amplification and PoNT devices.

  8. Comparsion of an immunochromatographic strip with ELISA for simultaneous detection of thiamphenicol, florfenicol and chloramphenicol in food samples.

    Science.gov (United States)

    Guo, Lingling; Song, Shanshan; Liu, Liqiang; Peng, Juan; Kuang, Hua; Xu, Chuanlai

    2015-09-01

    Rapid and sensitive indirect competitive enzyme-linked immunosorbent assays (ic-ELISA) and gold nanoparticle immunochromatographic strip tests were developed to detect thiamphenicol (TAP), florfenicol (FF) and chloramphenicol (CAP) in milk and honey samples. The generic monoclonal antibody for TAP, FF and CAP was prepared based on a hapten [D-threo-1-(4-aminophenyl)-2- dichloroacetylamino-1,3-propanediol], and the haptenwas linked to a carrier protein using the diazotization method. After the optimization of several parameters (coating, pH, sodium chloride content and methanol content), the ic-ELISA was established. The quantitative working range for TAP was 0.11-1.36 ng/mL, with an IC50 of 0.39 ng/mL. The optimized ELISA showed cross-reactivity to CAP (300%) and FF (15.6%), with IC50 values of 0.13 and 2.5 ng/mL, respectively. The analytical recovery of TAP, FF and CAP in milk and honey samples in the ic-ELISA ranged from 81.2 to 112.9%. Based on this monoclonal antibody, a rapid and sensitive immunochromatographic test strip was also developed. This strip had a detection limit of 1 ng/mL for TAP, FF and CAP in milk and honey samples. Moreover, the test was completed within 10 min. Our results showed that the proposed ic-ELISA and immunochromatographic test strip method are highly useful screening tools for TAP, FF and CAP detection in milk and honey samples. Copyright © 2015 John Wiley & Sons, Ltd.

  9. Simultaneous determination of methanol, acetaldehyde, acetone, and ethanol in human blood by gas chromatography with flame ionization detection.

    Science.gov (United States)

    Schlatter, J; Chiadmi, F; Gandon, V; Chariot, P

    2014-01-01

    Methanol, acetaldehyde, acetone, and ethanol, which are commonly used as biomarkers of several diseases, in acute intoxications, and forensic settings, can be detected and quantified in biological fluids. Gas chromatography (GC)-mass spectrometry techniques are complex, require highly trained personnel and expensive materials. Gas chromatographic determinations of ethanol, methanol, and acetone have been reported in one study with suboptimal accuracy. Our objective was to improve the assessment of these compounds in human blood using GC with flame ionization detection. An amount of 50 µl of blood was diluted with 300 µl of sterile water, 40 µl of 10% sodium tungstate, and 20 µl of 1% sulphuric acid. After centrifugation, 1 µl of the supernatant was injected into the gas chromatograph. We used a dimethylpolysiloxane capillary column of 30 m × 0.25 mm × 0.25 µm. We observed linear correlations from 7.5 to 240 mg/l for methanol, acetaldehyde, and acetone and from 75 to 2400 mg/l for ethanol. Precision at concentrations 15, 60, and 120 mg/l for methanol, acetaldehyde, and acetone and 150, 600, and 1200 mg/ml for ethanol were 0.8-6.9%. Ranges of accuracy were 94.7-98.9% for methanol, 91.2-97.4% for acetaldehyde, 96.1-98.7% for acetone, and 105.5-111.6% for ethanol. Limits of detection were 0.80 mg/l for methanol, 0.61 mg/l for acetaldehyde, 0.58 mg/l for acetone, and 0.53 mg/l for ethanol. This method is suitable for routine clinical and forensic practices.

  10. Simultaneous detection of the three ilarviruses affecting stone fruit trees by nonisotopic molecular hybridization and multiplex reverse-transcription polymerase chain reaction.

    Science.gov (United States)

    Saade, M; Aparicio, F; Sánchez-Navarro, J A; Herranz, M C; Myrta, A; Di Terlizzi, B; Pallás, V

    2000-12-01

    ABSTRACT The three most economically damaging ilarviruses affecting stone fruit trees on a worldwide scale are the related Prunus necrotic ringspot virus (PNRSV), Prune dwarf virus (PDV), and Apple mosaic virus (ApMV). Nonisotopic molecular hybridization and multiplex reverse-transcription polymerase chain reaction (RT-PCR) methodologies were developed that could detect all these viruses simultaneously. The latter technique was advantageous because it was discriminatory. For RT-PCR, a degenerate antisense primer was designed which was used in conjunction with three virus-specific sense primers. The amplification efficiencies for the detection of the three viruses in the multiplex RT-PCR reaction were identical to those obtained in the single RT-PCR reactions for individual viruses. This cocktail of primers was able to amplify sequences from all of the PNRSV, ApMV, and PDV isolates tested in five Prunus spp. hosts (almond, apricot, cherry, peach, and plum) occurring naturally in single or multiple infections. For ApMV isolates, differences in the electrophoretic mobilities of the PCR products were observed. The nucleotide sequence of the amplified products of two representative ApMV isolates was determined, and comparative analysis revealed the existence of a 28-nucleotide deletion in the sequence of isolates showing the faster electrophoretic mobility. To our knowledge, this is the first report on the simultaneous detection of three plant viruses by multiplex RT-PCR in woody hosts. This multiplex RT-PCR could be a useful time and cost saving method for indexing these three ilarviruses, which damage stone fruit tree yields, and for the analysis of mother plants in certification programs.

  11. Simultaneous determination of amiodarone and its metabolite desethylamiodarone by high-performance liquid chromatography with chemiluminescent detection

    International Nuclear Information System (INIS)

    Perez-Ruiz, Tomas; Martinez-Lozano, Carmen; Garcia-Martinez, Maria Dolores

    2008-01-01

    A novel method was developed for the determination of amiodarone and desethylamiodarone by high-performance liquid chromatography (HPLC) coupled with chemiluminescent (CL) detection. The procedure is based on the post-column photolysis of the analytes into photoproducts which are active in the tris(2,2'-bipyridyl)ruthenium(III) [Ru(bpy) 3 3+ ] CL system. Ru(bpy) 3 3+ was on-line generated by photo-oxidation of the Ru(II) complex in the presence of peroxydisulfate. The separation was carried out on a Mediterranea C 18 column with isocratic elution using a mixture of methanol and 0.017 mol L -1 ammonium sulfate buffer of pH 6.8. Under the optimum conditions, analytical curves, based on standard solutions, were linear over the range 0.1-50 μg mL -1 for amiodarone and 0.5-25 μg mL -1 for desethylamiodarone. The detection limits of amiodarone and desethylamiodarone were 0.02 and 0.11 μg mL -1 , respectively. Intra- and inter-day precision values of 0.9% relative standard deviation (R.S.D.) (n = 10) and 1.6% R.S.D. (n = 15), respectively, were obtained. The method was applied successfully to the determination of these compounds in serum and pharmaceutical formulations

  12. A PCR method targeting internal transcribed spacers: the simultaneous detection of Babesia bigemina and Babesia bovis in cattle.

    Science.gov (United States)

    Liu, Junlong; Guan, Guiquan; Liu, Aihong; Li, Youquan; Yin, Hong; Luo, Jianxun

    2014-03-01

    In this study, two pairs of oligonucleotide primers were designed according to the nucleotide sequence of the internal transcribed spacers (ITSs) of Babesia bigemina and B. bovis isolates from China. The primers were used in a multiplex PCR to detect parasite DNA in blood samples from cattle. There was no cross reactions with B. ovata, B. major, B. sp. Kashi, Theileria annulata, T. sergenti, T. sinensis or normal bovine DNA. The sensitivity of multiplex PCR assay was 1 pg and 10 pg DNA for B. bigemina and B. bovis, respectively. A total of 260 field blood samples collected from cattle in five provinces of China were analyzed by multiplex PCR and light microscopy. PCR testing revealed that 7.3% (19/260) and 5.8% (15/260) of cattle were positive for B. bigemina and B. bovis and 1.2% (3/260) of cattle were co-infected with B. bigemina and B. bovis. Using light microscopy, 2.3% (6/260) and 1.5% (4/260) of cattle were infected by B. bigemina and B. bovis, respectively, and no co-infection was found. The results showed that the multiplex PCR developed in the present study could be an alternative diagnostic tool for the detection of B. bovis and B. bigemina infection in cattle.

  13. Psychometric properties of the Need for Recovery after work scale: test-retest reliability and sensitivity to detect change

    NARCIS (Netherlands)

    de Croon, E. M.; Sluiter, J. K.; Frings-Dresen, M. H. W.

    2006-01-01

    BACKGROUND: Monitoring worker health and evaluating occupational healthcare interventions requires sensitive instruments that are reliable over time. The Need for Recovery scale (NFR), which quantifies workers' difficulties in recovering from work related exertions, may be a relevant instrument in

  14. The simultaneous ex vivo detection of low-frequency antigen-specific CD4+ and CD8+ T-cell responses using overlapping peptide pools.

    Science.gov (United States)

    Singh, Satwinder Kaur; Meyering, Maaike; Ramwadhdoebe, Tamara H; Stynenbosch, Linda F M; Redeker, Anke; Kuppen, Peter J K; Melief, Cornelis J M; Welters, Marij J P; van der Burg, Sjoerd H

    2012-11-01

    The ability to measure antigen-specific T cells at the single-cell level by intracellular cytokine staining (ICS) is a promising immunomonitoring tool and is extensively applied in the evaluation of immunotherapy of cancer. The protocols used to detect antigen-specific CD8+ T-cell responses generally work for the detection of antigen-specific T cells in samples that have undergone at least one round of in vitro pre-stimulation. Application of a common protocol but now using long peptides as antigens was not suitable to simultaneously detect antigen-specific CD8+ and CD4+ T cells directly ex vivo in cryopreserved samples. CD8 T-cell reactivity to monocytes pulsed with long peptides as antigens ranged between 5 and 25 % of that observed against monocytes pulsed with a direct HLA class I fitting minimal CTL peptide epitope. Therefore, we adapted our ICS protocol and show that the use of tenfold higher concentration of long peptides to load APC, the use of IFN-α and poly(I:C) to promote antigen processing and improve T-cell stimulation, does allow for the ex vivo detection of low-frequency antigen-specific CD8+ and CD4+ T cells in an HLA-independent setting. While most of the improvements were related to increasing the ability to measure CD8+ T-cell reactivity following stimulation with long peptides to at least 50 % of the response detected when using a minimal peptide epitope, the final analysis of blood samples from vaccinated patients successfully showed that the adapted ICS protocol also increases the ability to ex vivo detect low-frequency p53-specific CD4+ T-cell responses in cryopreserved PBMC samples.

  15. Simultaneous detection for three kinds of veterinary drugs: Chloramphenicol, clenbuterol and 17-beta-estradiol by high-throughput suspension array technology

    Energy Technology Data Exchange (ETDEWEB)

    Liu Nan; Su Pu [Institute of Hygiene and Environmental Medicine, Tianjin 300050 (China); Gao Zhixian [Institute of Hygiene and Environmental Medicine, Tianjin 300050 (China)], E-mail: gaozhx@163.com; Zhu Maoxiang; Yang Zhihua; Pan Xiujie [Institute of Radiation Medicine, Beijing 100850 (China); Fang Yanjun; Chao Fuhuan [Institute of Hygiene and Environmental Medicine, Tianjin 300050 (China)

    2009-01-19

    Suspension array technology for simultaneous detection of three kinds of veterinary drugs, chloramphenicol (CAP), clenbuterol and 17-beta-estradiol has been developed. Conjugates of chloramphenicol and clenbuterol coupled with bovine serum albumin were synthesized and purified. Probes of suspension array were constituted by coupling the three conjugates on the fluorescent microspheres/beads and the microstructures of the beads' surface were observed by scanning electron microscopy which was a direct confirmation for the successful conjugates' coupling. The optimal addition of conjugates and the amounts of antibodies were optimized and selected, respectively. Standard curves were plotted and the coefficient of determination-R{sup 2} was greater than 0.989 which suggested good logistic correlation. The detection ranges for the three veterinary drugs are 40-6.25 x 10{sup 5} ng L{sup -1}, 50-7.81 x 10{sup 5} ng L{sup -1} and 1 x 10{sup 3-}7.29 x 10{sup 5} ng L{sup -1}, respectively and the lowest detection limits (LDLs) of them are 40, 50 and 1000 ng L{sup -1}, respectively. The suspension array is specific and has no significant cross-reactivity with other chemicals. Meanwhile, unknown samples were detected by suspension array and ELISA in comparison with each other. The errors between found and real for the detection of the unknown samples were relatively small to both of the two methods, whereas, the detection ranges of suspension array are broader and sensitive than that of the traditional ELISA. The high-throughput suspension array is proved to be a novel method for multi-analysis of veterinary drugs with simple operation, high sensitivity and low cost.

  16. Simultaneous detection of Acidovorax avenae subsp. citrulli and Didymella bryoniae in cucurbit seedlots using magnetic capture hybridization and real-time polymerase chain reaction.

    Science.gov (United States)

    Ha, Y; Fessehaie, A; Ling, K S; Wechter, W P; Keinath, A P; Walcott, R R

    2009-06-01

    To improve the simultaneous detection of two pathogens in cucurbit seed, a combination of magnetic capture hybridization (MCH) and multiplex real-time polymerase chain reaction (PCR) was developed. Single-stranded DNA hybridization capture probes targeting DNA of Acidovorax avenae subsp. citrulli, causal agent of bacterial fruit blotch, and Didymella bryoniae, causal agent of gummy stem blight, were covalently attached to magnetic particles and used to selectively concentrate template DNA from cucurbit seed samples. Sequestered template DNAs were subsequently amplified by multiplex real-time PCR using pathogen-specific TaqMan PCR assays. The MCH multiplex real-time PCR assay displayed a detection threshold of A. avenae subsp. citrulli at 10 CFU/ml and D. bryoniae at 10(5) conidia/ml in mixtures of pure cultures of the two pathogens, which was 10-fold more sensitive than the direct real-time PCR assays for the two pathogens separately. Although the direct real-time PCR assay displayed a detection threshold for A. avenae subsp. citrulli DNA of 100 fg/microl in 25% (1/4 samples) of the samples assayed, MCH real-time PCR demonstrated 100% detection frequency (4/4 samples) at the same DNA concentration. MCH did not improve detection sensitivity for D. bryoniae relative to direct real-time PCR using conidial suspensions or seed washes from D. bryoniae-infested cucurbit seed. However, MCH real-time PCR facilitated detection of both target pathogens in watermelon and melon seed samples (n = 5,000 seeds/sample) in which 0.02% of the seed were infested with A. avenae subsp. citrulli and 0.02% were infested with D. bryoniae.

  17. Development of degenerate and species-specific primers for the differential and simultaneous RT-PCR detection of grapevine-infecting nepoviruses of subgroups A, B and C.

    Science.gov (United States)

    Digiaro, Michele; Elbeaino, Toufic; Martelli, Giovanni Paolo

    2007-04-01

    Based on the nucleotide sequence homology of RNA-1 and RNA-2 of nepoviruses isolated from grapevines, three sets of degenerate primers, one for each of the three subgroups of the genus (A, B and C), were designed and proved effective for RT-PCR detection of subgroups in infected grapevines and herbaceous hosts. Primers designed specifically for detecting subgroup A species amplified a fragment of 255 bp from samples infected by Grapevine fanleaf virus (GFLV), Arabis mosaic virus (ArMV), Tobacco ringspot virus (TRSV) and Grapevine deformation virus (GDefV), but not from samples infected by other nepovirus species. Similarly, primers for detection of subgroup B nepoviruses amplified a 390 bp product from samples infected by Grapevine chrome mosaic virus (GCMV), Tomato black ring virus (TBRV), Grapevine Anatolian ringspot virus (GARSV) and Artichoke Italian latent virus (AILV). The third set of primers amplified a 640 bp fragment, only from samples infected by subgroup C nepoviruses, i.e Tomato ringspot virus (ToRSV) Grapevine Bulgarian latent virus (GBLV), and Grapevine Tunisian ringspot virus (GTRSV). These primers were able to detect simultaneously all viral species belonging to the same subgroup and to discriminate species of different subgroups. Multiplex-PCR detection of subgroup A and B nepoviruses was obtained using a specific primer (sense for subgroup A and antisense for subgroup B) for each of the species of the same subgroup in combination with the degenerate subgroup-specific primers. In this way it was possible to detect four different viral species in single samples containing mixtures of viruses of the same subgroup. In particular, for viruses of subgroup A (TRSV, GFLV, ArMV and GDefV) amplicons of 190, 259, 301 and 371 bp were obtained, whereas amplicons of 190, 278, 425 and 485 bp, respectively, were obtained from samples infected with viruses of subgroup B (GCMV, AILV, GARSV and TBRV).

  18. The First Simultaneous X-Ray/Radio Detection of the First Be/BH System MWC 656

    Energy Technology Data Exchange (ETDEWEB)

    Ribó, M.; Paredes, J. M.; Marcote, B.; Moldón, J.; Paredes-Fortuny, X. [Departament de Física Quàntica i Astrofísica, Institut de Ciències del Cosmos (ICCUB), Universitat de Barcelona, IEEC-UB, Martí i Franquès 1, E08028 Barcelona (Spain); Munar-Adrover, P. [INAF/IAPS-Roma, I-00133 Roma (Italy); Iwasawa, K. [ICREA, Institut de Ciències del Cosmos (ICCUB), Universitat de Barcelona, IEEC-UB, Martí i Franquès 1, E-08028 Barcelona (Spain); Casares, J. [Instituto de Astrofísica de Canarias, E-38200 La Laguna, Tenerife (Spain); Migliari, S. [European Space Astronomy Centre, Apartado/P.O. Box 78, Villanueva de la Canada, E-28691 Madrid (Spain)

    2017-02-01

    MWC 656 is the first known Be/black hole (BH) binary system. Be/BH binaries are important in the context of binary system evolution and sources of detectable gravitational waves because they are possible precursors of coalescing neutron star/BH binaries. X-ray observations conducted in 2013 revealed that MWC 656 is a quiescent high-mass X-ray binary (HMXB), opening the possibility to explore X-ray/radio correlations and the accretion/ejection coupling down to low luminosities for BH HMXBs. Here we report on a deep joint Chandra /VLA observation of MWC 656 (and contemporaneous optical data) conducted in 2015 July that has allowed us to unambiguously identify the X-ray counterpart of the source. The X-ray spectrum can be fitted with a power law with Γ ∼ 2, providing a flux of ≃4 × 10{sup −15} erg cm{sup −2} s{sup −1} in the 0.5–8 keV energy range and a luminosity of L {sub X} ≃ 3 × 10{sup 30} erg s{sup −1} at a 2.6 kpc distance. For a 5 M{sub ⊙} BH this translates into ≃5 × 10{sup −9} L {sub Edd}. These results imply that MWC 656 is about 7 times fainter in X-rays than it was two years before and reaches the faintest X-ray luminosities ever detected in stellar-mass BHs. The radio data provide a detection with a peak flux density of 3.5 ± 1.1 μ Jy beam{sup −1}. The obtained X-ray/radio luminosities for this quiescent BH HMXB are fully compatible with those of the X-ray/radio correlations derived from quiescent BH low-mass X-ray binaries. These results show that the accretion/ejection coupling in stellar-mass BHs is independent of the nature of the donor star.

  19. Performance of a polymer coated silicon microarray for simultaneous detection of food allergen-specific IgE and IgG4.

    Science.gov (United States)

    Sievers, S; Cretich, M; Gagni, P; Ahrens, B; Grishina, G; Sampson, H A; Niggemann, B; Chiari, M; Beyer, K

    2017-08-01

    Microarray-based component-resolved diagnostics (CRD) has become an accepted tool to detect allergen-specific IgE sensitization towards hundreds of allergens in parallel from one drop of serum. Nevertheless, specificity and sensitivity as well as a simultaneous detection of allergen-specific IgG 4 , as a potential parameter for tolerance development, remain to be optimized. We applied the recently introduced silicon chip coated with a functional polymer named copoly(DMA-NAS-MAPS) to the simultaneous detection of food allergen-specific IgE and IgG 4 , and compared it with ImmunoCAP and ImmunoCAP ISAC. Inter- and intraslide variation, linearity of signal and working range, sensitivity and application of internal calibrations for IgE and IgG 4 were assessed. Native and recombinant allergenic proteins from hen's egg and cow's milk were spotted on silicon chips coated with copoly(DMA-NAS-MAPS) along with known concentrations for human IgE and IgG 4 . A serum pool and 105 patient samples were assessed quantitatively and semi-quantitatively with the ImmunoCAP and ImmunoCAP ISAC and correlated with IgE- and IgG 4 -specific fluorescence on silicon microarrays. Allergen-specific IgE and IgG 4 were detected in parallel using two fluorescent dyes with no crosstalk. Results from the ImmunoCAP correlated better with microarray fluorescence than with ImmunoCAP ISAC except for the allergen ovomucoid. The working range of the silicon microarray for total hen's egg-specific IgE was comparable to the range of 0.1 to >100 kU A /L of the ImmunoCAP system, whereas for total cow's milk, the silicon microarray was less sensitive. Detectable allergen-specific IgG 4 could be determined only for low concentrations, but still correlated positively with ImmunoCAP results. We confirmed the ability of the polymer coated silicon microarray to be comparably sensitive to the ImmunoCAP ISAC for various food allergens. This suggests that the copoly(DMA-NAS-MAPS) microarray is a low-cost, self

  20. Simultaneous determination of nucleosides, myriocin, and carbohydrates in Cordyceps by HPLC coupled with diode array detection and evaporative light scattering detection.

    Science.gov (United States)

    Wang, Shuang; Yang, Feng-Qing; Feng, Kun; Li, De-Qiang; Zhao, Jing; Li, Shao-Ping

    2009-12-01

    A HPLC coupled with diode array detection (DAD) and evaporative light scattering detection (ELSD) method for qualitative and quantitative analysis of eight nucleosides and nucleobases, three carbohydrates and myriocin in Cordyceps was developed. A Prevail Carbohydrate ES column was employed for the separation within 50 min. Nucleosides and their bases were tested at UV 254 nm. ELSD was connected with DAD to determine myriocin and carbohydrates. The optimum drift tube temperature of ELSD was at 94 degrees C with the nitrogen flow rate of 2.0 L/min. All calibration curves showed good linearity (R(2)>0.9933) during the test ranges. The precision, repeatability, accuracy, LOD and LOQ were also fully investigated. This developed method was successfully applied to quantify 12 components, eight nucleosides and nucleobases, three carbohydrates and myriocin, in natural and cultured Cordyceps, which provides another view for quality control of Cordyceps sinensis.

  1. Monolithic silica spin column extraction and simultaneous derivatization of amphetamines and 3,4-methylenedioxyamphetamines in human urine for gas chromatographic-mass spectrometric detection

    International Nuclear Information System (INIS)

    Nakamoto, Akihiro; Nishida, Manami; Saito, Takeshi; Kishiyama, Izumi; Miyazaki, Shota; Murakami, Katsunori; Nagao, Masataka; Namura, Akira

    2010-01-01

    A simple, sensitive, and specific method with gas chromatography-mass spectrometry was developed for simultaneous extraction and derivatization of amphetamines (APs) and 3,4-methylenedioxyamphetamines (MDAs) in human urine by using a monolithic silica spin column. All the procedures, such as sample loading, washing, and elution were performed by centrifugation. APs and MDAs in urine were adsorbed on the monolithic silica and derivatized with propyl chloroformate in the column. Methamphetamine-d 5 was used as an internal standard. The linear ranges were 0.01-5.0 μg mL -1 for methamphetamine (MA) and 3,4-methylenedioxymethamphetamine (MDMA) and 0.02-5.0 μg mL -1 for amphetamine (AP) and 3,4-methylenedioxyamphetamine (MDA) (coefficient of correlation ≥0.995). The recovery of APs and MDAs in urine was 84-94%, and the relative standard deviation of the intra- and interday reproducibility for urine samples containing 0.1, 1.0, and 4.0 μg mL -1 of APs and MDAs ranged from 1.4% to 13.6%. The lowest detection limit (signal-to-noise ratio ≥ 3) in urine was 5 ng mL -1 for MA and MDMA and 10 ng mL -1 for AP and MDA. The proposed method can be used to perform simultaneous extraction and derivatization on spin columns that have been loaded with a small quantity of solvent by using centrifugation.

  2. SU-G-201-03: Automation of High Dose Rate Brachytherapy Quality Assurance: Development of a Radioluminescent Detection System for Simultaneous Detection of Activity, Timing, and Positioning

    Energy Technology Data Exchange (ETDEWEB)

    Jenkins, C; Xing, L; Fahimian, B [Stanford University, Stanford, CA (United States)

    2016-06-15

    Purpose: Accuracy of positioning, timing and activity is of critical importance for High Dose Rate (HDR) brachytherapy delivery. Respective measurements via film autoradiography, stop-watches and well chambers can be cumbersome, crude or lack dynamic source evaluation capabilities. To address such limitations, a single device radioluminescent detection system enabling automated real-time quantification of activity, position and timing accuracy is presented and experimentally evaluated. Methods: A radioluminescent sheet was fabricated by mixing Gd?O?S:Tb with PDMS and incorporated into a 3D printed device where it was fixated below a CMOS digital camera. An Ir-192 HDR source (VS2000, VariSource iX) with an effective active length of 5 mm was introduced using a 17-gauge stainless steel needle below the sheet. Pixel intensity values for determining activity were taken from an ROI centered on the source location. A calibration curve relating intensity values to activity was generated and used to evaluate automated activity determination with data gathered over 6 weeks. Positioning measurements were performed by integrating images for an entire delivery and fitting peaks to the resulting profile. Timing measurements were performed by evaluating source location and timestamps from individual images. Results: Average predicted activity error over 6 weeks was .35 ± .5%. The distance between four dwell positions was determined by the automated system to be 1.99 ± .02 cm. The result from autoradiography was 2.00 ± .03 cm. The system achieved a time resolution of 10 msec and determined the dwell time to be 1.01 sec ± .02 sec. Conclusion: The system was able to successfully perform automated detection of activity, positioning and timing concurrently under a single setup. Relative to radiochromic and radiographic film-based autoradiography, which can only provide a static evaluation positioning, optical detection of temporary radiation induced luminescence enables dynamic

  3. Simultaneous and rapid determination of caffeine and taurine in energy drinks by MEKC in a short capillary with dual contactless conductivity/photometry detection.

    Science.gov (United States)

    Vochyánová, Blanka; Opekar, František; Tůma, Petr

    2014-06-01

    A method has been developed for the simultaneous determination of taurine and caffeine using a laboratory-made instrument enabling separation analysis in a short 10.5 cm capillary. The substances are detected using a contactless conductometry/ultraviolet (UV) photometry detector that enables recording both signals at one place in the capillary. The separation of caffeine and taurine was performed using the MEKC technique in a BGE with the composition 40 mM CHES, 15 mM NaOH, and 50 mM SDS, pH 9.36. Under these conditions, the migration time of caffeine is 43 s and of taurine 60 s; LOD for caffeine is 4 mg/L using photometric detection and LOD for taurine is 24 mg/L using contactless conductometric detection. The standard addition method was used for determination in Red Bull energy drink of caffeine 317 mg/L and taurine 3860 mg/L; the contents in Kamikaze drink were 468 mg/L caffeine and 4110 mg/L taurine. The determined values are in good agreement with the declared contents of these substances. RSD does not exceed 3%. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. The G-BHQ synergistic effect: Improved double quenching molecular beacons based on guanine and Black Hole Quencher for sensitive simultaneous detection of two DNAs.

    Science.gov (United States)

    Xiang, Dongshan; Li, Fengquan; Wu, Chenyi; Shi, Boan; Zhai, Kun

    2017-11-01

    We designed two double quenching molecular beacons (MBs) with simple structure based on guanine (G base) and Black Hole Quencher (BHQ), and developed a new analytical method for sensitive simultaneous detection of two DNAs by synchronous fluorescence analysis. In this analytical method, carboxyl fluorescein (FAM) and tetramethyl-6-carboxyrhodamine (TAMRA) were respectively selected as fluorophore of two MBs, Black Hole Quencher 1 (BHQ-1) and Black Hole Quencher 2 (BHQ-2) were respectively selected as organic quencher, and three continuous nucleotides with G base were connected to organic quencher (BHQ-1 and BHQ-2). In the presence of target DNAs, the two MBs hybridize with the corresponding target DNAs, the fluorophores are separated from organic quenchers and G bases, leading to recovery of fluorescence of FAM and TAMRA. Under a certain conditions, the fluorescence intensities of FAM and TAMRA all exhibited good linear dependence on their concentration of target DNAs (T1 and T2) in the range from 4 × 10 -10 to 4 × 10 -8 molL -1 (M). The detection limit (3σ, n = 13) of T1 was 3 × 10 -10 M and that of T2 was 2×10 -10 M, respectively. Compared with the existing analysis methods for multiplex DNA with MBs, this proposed method based on double quenching MBs is not only low fluorescence background, short analytical time and low detection cost, but also easy synthesis and good stability of MB probes. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Avoidance Behavior against Positive Allergens Detected with a Multiple Allergen Simultaneous Test Immunoblot Assay in Patients with Urticaria: Factors Associated with Avoidance Success/Failure.

    Science.gov (United States)

    Lee, Min Kyung; Kwon, In Ho; Kim, Han Su; Kim, Heung Yeol; Cho, Eun Byul; Bae, Youin; Park, Gyeong Hun; Park, Eun Joo; Kim, Kwang Ho; Kim, Kwang Joong

    2016-02-01

    Avoidance behavior against positive allergens detected by using multiple allergen simultaneous test (MAST)-immunoblot assay in patients with urticaria has been rarely reported. We aimed to assess the avoidance behavior of patients with urticaria against positive allergens detected with a MAST. One hundred and one urticaria patients who showed positivity to at least one allergen on a MAST completed a questionnaire regarding their test results. The avoidance behavior of the patients was evaluated, and relevant determining factors of avoidance success/failure were statistically assessed. We detected 144 different data (n=51, food allergens; n=17, pollen allergens; and n=76, aeroallergens) from 101 patients with urticaria. The avoidance failure rates were 33.3% for food allergens, 70.6% for pollen allergens, and 30.3% for aeroallergens. The pollen group showed a significantly higher avoidance failure rate than the food and aeroallergen groups (psuccessfully avoid allergens (psuccess or failure against allergens in patients with urticaria when clinicians conduct allergen-specific immunoglobulin E tests.

  6. Simultaneous determination of low-molecular-weight organic acids and chlorinated acid herbicides in environmental water by a portable CE system with contactless conductivity detection.

    Science.gov (United States)

    Xu, Yan; Wang, Weilong; Li, Sam Fong Yau

    2007-05-01

    This report describes a method to simultaneously determine 11 low-molecular-weight (LMW) organic acids and 16 chlorinated acid herbicides within a single run by a portable CE system with contactless conductivity detection (CCD) in a poly(vinyl alcohol) (PVA)-coated capillary. Under the optimized condition, the LODs of CE-CCD ranged from 0.056 to 0.270 ppm, which were better than for indirect UV (IUV) detection of the 11 LMW organic acids or UV detection of the 16 chlorinated acid herbicides. Combined with an on-line field-amplified sample stacking (FASS) procedure, sensitivity enhancement of 632- to 1078-fold was achieved, with satisfactory reproducibility (RSDs of migration times less than 2.2%, and RSDs of peak areas less than 5.1%). The FASS-CE-CCD method was successfully applied to determine the two groups of acidic pollutants in two kinds of environmental water samples. The portable CE-CCD system shows advantages such as simplicity, cost effectiveness, and miniaturization. Therefore, the method presented in this report has great potential for onsite analysis of various pollutants at the trace level.

  7. Implementation of a SPR immunosensor for the simultaneous detection of the 22K and 20K hGH isoforms in human serum samples.

    Science.gov (United States)

    de Juan-Franco, Elena; Rodríguez-Frade, J M; Mellado, M; Lechuga, Laura M

    2013-09-30

    We have implemented a Surface Plasmon Resonance (SPR) immunosensor based on a sandwich assay for the simultaneous detection of the two main hGH isoforms, of 22 kDa (22K) and 20 kDa (20K). An oriented-antibody sensor surface specific for both hormone isoforms was assembled by using the biotin-streptavidin system. The immunosensor functionality was checked for the direct detection of the 22K hGH isoform in buffer, which gave high specificity and reproducibility (intra and inter-assay mean coefficients of variation of 8.23% and 9% respectively). The selective determination of the 22K and 20K hGH isoforms in human serum samples in a single assay was possible by using two specific anti-hGH monoclonal antibodies. The detection limit for both hormone isoforms was 0.9 ng mL(-1) and the mean coefficient of variation was below 7.2%. The excellent reproducibility and sensitivity obtained indicate the high performance of this immunosensor for implementing an anti-doping test. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Simultaneous Automatic Electrochemical Detection of Zinc, Cadmium, Copper and Lead Ions in Environmental Samples Using a Thin-Film Mercury Electrode and an Artificial Neural Network

    Directory of Open Access Journals (Sweden)

    Jiri Kudr

    2014-12-01

    Full Text Available In this study a device for automatic electrochemical analysis was designed. A three electrodes detection system was attached to a positioning device, which enabled us to move the electrode system from one well to another of a microtitre plate. Disposable carbon tip electrodes were used for Cd(II, Cu(II and Pb(II ion quantification, while Zn(II did not give signal in this electrode configuration. In order to detect all mentioned heavy metals simultaneously, thin-film mercury electrodes (TFME were fabricated by electrodeposition of mercury on the surface of carbon tips. In comparison with bare electrodes the TMFEs had lower detection limits and better sensitivity. In addition to pure aqueous heavy metal solutions, the assay was also performed on mineralized rock samples, artificial blood plasma samples and samples of chicken embryo organs treated with cadmium. An artificial neural network was created to evaluate the concentrations of the mentioned heavy metals correctly in mixture samples and an excellent fit was observed (R2 = 0.9933.

  9. A validated high-performance liquid chromatography method with diode array detection for simultaneous determination of nine flavonoids in Senecio cannabifolius Less.

    Science.gov (United States)

    Niu, Tian-Zeng; Zhang, Yu-Wei; Bao, Yong-Li; Wu, Yin; Yu, Chun-Lei; Sun, Lu-Guo; Yi, Jing-Wen; Huang, Yan-Xin; Li, Yu-Xin

    2013-03-25

    A reversed phase high performance liquid chromatography method coupled with a diode array detector (HPLC-DAD) was developed for the first time for the simultaneous determination of 9 flavonoids in Senecio cannabifolius, a traditional Chinese medicinal herb. Agilent Zorbax SB-C18 column was used at room temperature and the mobile phase was a mixture of acetonitrile and 0.5% formic acid (v/v) in water in the gradient elution mode at a flow-rate of 1.0mlmin(-1), detected at 360nm. Validation of this method was performed to verify the linearity, precision, limits of detection and quantification, intra- and inter-day variabilities, reproducibility and recovery. The calibration curves showed good linearities (R(2)>0.9995) within the test ranges. The relative standard deviation (RSD) of the method was less than 3.0% for intra- and inter-day assays. The samples were stable for at least 96h, and the average recoveries were between 90.6% and 102.5%. High sensitivity was demonstrated with detection limits of 0.028-0.085μg/ml for flavonoids. The newly established HPLC method represents a powerful technique for the quality assurance of S. cannabifolius. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Simultaneous determination of 13 carbohydrates using high-performance anion-exchange chromatography coupled with pulsed amperometric detection and mass spectrometry.

    Science.gov (United States)

    Zhao, Dan; Feng, Feng; Yuan, Fei; Su, Jin; Cheng, Yan; Wu, Hanqiu; Song, Kun; Nie, Bo; Yu, Lian; Zhang, Feng

    2017-04-01

    A simple, accurate, and highly sensitive method was developed for the determination of 13 carbohydrates in polysaccharide of Spirulina platensis based on high-performance anion-exchange chromatography coupled with pulsed amperometric detection and mass spectrometry. Samples were extracted with deionized water using ultrasonic-assisted extraction, and the ultrasound-assisted extraction conditions were optimized by Box-Behnken design. Then the extracted polysaccharide was hydrolyzed by adding 1 mol/L trifluoroacetic acid before determination by high-performance anion-exchange chromatography coupled with pulsed amperometric detection and confirmed by high-performance anion-exchange chromatography coupled with mass spectrometry. The high-performance anion-exchange chromatography coupled with pulsed amperometric detection method was performed on a CarboPac PA20 column by gradient elution using deionized water, 0.1 mol/L sodium hydroxide solution, and 0.4 mol/L sodium acetate solution. Excellent linearity was observed in the range of 0.05-10 mg/L. The average recoveries ranged from 80.7 to 121.7%. The limits of detection and limits of quantification for 13 carbohydrates were 0.02-0.10 and 0.2-1.2  μg/kg, respectively. The developed method has been successfully applied to ambient samples, and the results indicated that high-performance anion-exchange chromatography coupled with pulsed amperometric detection and mass spectrometry could provide a rapid and accurate method for the simultaneous determination of carbohydrates. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Simultaneous analysis of the expression of 14 genes with individual prognostic value in myelodysplastic syndrome patients at diagnosis: WT1 detection in peripheral blood adversely affects survival.

    Science.gov (United States)

    Santamaría, Carlos; Ramos, Fernando; Puig, Noemi; Barragán, Eva; de Paz, Raquel; Pedro, Carme; Insunza, Andrés; Tormo, Mar; Del Cañizo, Consuelo; Diez-Campelo, María; Xicoy, Blanca; Salido, Eduardo; Sánchez del Real, Javier; Hernández, Montserrat; Chillón, Carmen; Sanz, Guillermo F; García-Sanz, Ramón; San Miguel, Jesús F; González, Marcos

    2012-12-01

    Several studies have evaluated the prognostic value of the individual expression of certain genes in patients with myelodysplastic syndromes (MDS). However, none of them includes their simultaneous analysis by quantitative polymerase chain reaction (PCR). We evaluated relative expression levels of 14 molecular markers in 193 peripheral blood samples from untreated MDS patients using real-time PCR. Detectable WT1 expression levels, low TET2, and low IER3 gene expression were the only markers showing in univariate analysis a poor prognostic value for all treatment-free (TFS), progression-free (PFS), and overall survival (OS). In multivariate analysis, molecular parameters associated with a shorter TFS were: WT1 detection (p = 0.014), low TET2 (p = 0.002), and low IER3 expression (p = 0.025). WT1 detection (p = 0.006) and low TET2 (p = 0.006) expression were associated with a shorter PFS when multivariate analysis was carried out by including only molecular markers. Molecular values with an independent value in OS were: WT1 detection (p = 0.003), high EVI1 expression (p = 0.001), and undetectatable p15-CDKN2B (p = 0.037). WT1 expressers were associated with adverse clinical-biological features, high IPSS and WPSS scoring, and unfavorable molecular expression profile. In summary, detectable WT1 expression levels, and low TET2 and low IER3 expression in peripheral blood showed a strong association with adverse prognosis in MDS patients at diagnosis. However, WT1 was the only molecular marker displaying an independent prognostic value in both OS and TFS.

  12. Time-dependent exchange and tunneling: detection at the same place of two electrons emitted simultaneously from different sources

    International Nuclear Information System (INIS)

    Marian, D; Colomés, E; Oriols, X

    2015-01-01

    Two-particle scattering probabilities in tunneling scenarios with exchange interaction are analyzed with quasi-particle wave packets. Two initial one-particle wave packets (with opposite central momentums) are spatially localized at each side of a barrier. After impinging upon a tunneling barrier, each wave packet splits into transmitted and reflected components. When the initial two-particle anti-symmetrical state is defined as a Slater determinant of any type of (normalizable) one-particle wave packet, it is shown that the probability of detecting two (identically injected) electrons at the same side of the barrier is different from zero in very common (single or double barrier) scenarios. In some particular scenarios, the transmitted and reflected components become orthogonal and the mentioned probabilities reproduce those values associated to distinguishable particles. These unexpected non-zero probabilities are still present when non-separable Coulomb interaction or non-symmetrical potentials are considered. On the other hand, for initial wave packets close to Hamiltonian eigenstates, the usual zero two-particle probability for electrons at the same side of the barrier found in the literature is recovered. The generalization to many-particle scattering probabilities with quasi-particle wave packets for low and high phase-space density are also analyzed. The far-reaching consequences of these non-zero probabilities in the accurate evaluation of quantum noise in mesoscopic systems are briefly indicated. (paper)

  13. Simultaneous Determination of Six Food Additives in drinks by high performance liquid chromatography coupled to diode array detector detection

    International Nuclear Information System (INIS)

    Yan, Q.

    2013-01-01

    A reversed-phase high performance liquid chromatographic method for the successful separation and determination of 6 synthetic food additives (aspartame, acesulfame potassium, benzoic acid, sodium saccharin, tartrazine and sunset yellow) was developed. A EclipseXDB-C18 column (250x4.6 mm I.D.; 5 micro m) was used and the mobile phase contained methanol and 0.02 mol/L ammonium acetate (pH 6.0) (30:70, v/v) was pumped at a flow rate of 0.7 mL/min at room temperature. Successful separation conditions were obtained for all the compounds using an optimized gradient elution within 10 min. The diode array detector was used to monitor the food additives at 230 nm. The method was thoroughly validated, detection limits for all substances varied between 0.03 and 1.35 micro g/mg, the intra-day precision (as RSD) ranged from 1.57% to 4.72 %, the inter-day precision (as RSD) was between 2.05 % and 4.18 %. Satisfactory recoveries, ranging from 90.00 % to 109.87 %, were obtained. The proposed system was applied to drink samples. (author)

  14. Simultaneous detection of Staphylococcus aureus and Salmonella typhimurium using multicolor time-resolved fluorescence nanoparticles as labels.

    Science.gov (United States)

    Wang, Xiaole; Huang, Yukun; Wu, Shijia; Duan, Nuo; Xu, Baocai; Wang, Zhouping

    2016-11-21

    Foodborne illnesses caused by Staphylococcus aureus and Salmonella typhimurium are common public health issues worldwide, affecting both developing and developed countries. In this study, aptamers labeled with multicolor lanthanide-doped time-resolved fluorescence (TRFL) nanoparticles were used as signal probes, and immobilized by Fe 3 O 4 magnetic nanoparticles were used as the capture probes. The signal probes were bonded onto the captured bacteria by the recognition of aptamer to form the sandwich-type complex. Under the optimal conditions, TRFL intensity at 544nm was used to quantify S. typhimurium (y=10,213×-12,208.92, R 2 =0.9922) and TRFL intensity at 615nm for S. aureus (y=4803.20×-1933.87, R 2 =0.9982) in the range of 10 2 -10 5 CFU/ml. Due to the magnetic separation and concentration of Fe 3 O 4 nanoparticles, detection limits of the developed method were found to be 15, 20CFU/ml for S. typhimurium and S. aureus, respectively. The application of this bioassay in milk was also investigated, and results were consistent with those of plate-counting method. Therefore, this simple and rapid method owns a great potential in the application for the multiplex analysis in food safety. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Application of the FICTION technique for the simultaneous detection of immunophenotype and chromosomal abnormalities in routinely fixed, paraffin wax embedded bone marrow trephines

    Science.gov (United States)

    Korać, P; Jones, M; Dominis, M; Kušec, R; Mason, D Y; Banham, A H; Ventura, R A

    2005-01-01

    The use of interphase fluorescence in situ hybridisation (FISH) to study cytogenetic abnormalities in routinely fixed paraffin wax embedded tissue has become commonplace over the past decade. However, very few studies have applied FISH to routinely fixed bone marrow trephines (BMTs). This may be because of the acid based decalcification methods that are commonly used during the processing of BMTs, which may adversely affect the suitability of the sample for FISH analysis. For the first time, this report describes the simultaneous application of FISH and immunofluorescent staining (the FICTION technique) to formalin fixed, EDTA decalcified and paraffin wax embedded BMTs. This technique allows the direct correlation of genetic abnormalities to immunophenotype, and therefore will be particularly useful for the identification of genetic abnormalities in specific tumour cells present in BMTs. The application of this to routine clinical practice will assist diagnosis and the detection of minimal residual disease. PMID:16311361

  16. Ascorbic acid surface modified TiO2-thin layers as a fully integrated analysis system for visual simultaneous detection of organophosphorus pesticides

    Science.gov (United States)

    Li, Shunxing; Liang, Wenjie; Zheng, Fengying; Lin, Xiaofeng; Cai, Jiabai

    2014-11-01

    TiO2 photocatalysis and colorimetric detection are coupled with thin layer chromatography (TLC) for the first time to develop a fully integrated analysis system. Titania@polystyrene hybrid microspheres were surface modified with ascorbic acid, denoted AA-TiO2@PS, and used as the stationary phase for TLC. Because the affinity between AA-TiO2@PS and organophosphorus pesticides (OPs) was different for different species of OPs (including chlopyrifos, malathion, parathion, parathion-methyl, and methamidophos), OPs could be separated simultaneously by the mobile phase in 12.0 min with different Rf values. After surface modification, the UV-vis wavelength response range of AA-TiO2@PS was expanded to 650 nm. Under visible-light irradiation, all of the OPs could be photodegraded to PO43- in 25.0 min. Based on the chromogenic reaction between PO43- and chromogenic agents (ammonium molybdate and ascorbic acid), OPs were quantified from color intensity images using a scanner in conjunction with image processing software. So, AA-TiO2@PS was respectively used as the stationary phase of TLC for efficient separation of OPs, as a photocatalyst for species transformation of phosphorus, and as a colorimetric probe for on-field simultaneous visual detection of OPs in natural water. Linear calibration curves for each OP ranged from 19.3 nmol P L-1 to 2.30 μmol P L-1. This integrated analysis system was simple, inexpensive, easy to operate, and sensitive.TiO2 photocatalysis and colorimetric detection are coupled with thin layer chromatography (TLC) for the first time to develop a fully integrated analysis system. Titania@polystyrene hybrid microspheres were surface modified with ascorbic acid, denoted AA-TiO2@PS, and used as the stationary phase for TLC. Because the affinity between AA-TiO2@PS and organophosphorus pesticides (OPs) was different for different species of OPs (including chlopyrifos, malathion, parathion, parathion-methyl, and methamidophos), OPs could be separated

  17. Reliable Maintanace of Wireless Sensor Networks for Event-detection Applications%事件检测型传感器网络的可靠性维护

    Institute of Scientific and Technical Information of China (English)

    胡四泉; 杨金阳; 王俊峰

    2011-01-01

    The reliability maintannace of the wireless sensor network is a key point to keep the alarm messages delivered reliably to the monitor center on time in a event-detection application. Based on the unreliable links in the wireless sensor network and the network charateristics of an event detection application,MPRRM,a multiple path redundant reliability maintanace algoritm was proposed in this paper. Both analytical and simulation results show that the MPRRM algorithm is superior to the previous published solutions in the metrics of reliability, false positive rate, latency and message overhead.%传感器网络(Wireless Sensor Networks,WSN)的事件检测型应用中,如何通过可靠性维护来保证在检测到事件时报警信息能及时、可靠地传输到监控主机至关重要.通过对不可靠的无线链路和网络传输的分析,提出多路冗余可靠性维护算法MPRRM.通过解析方法和仿真分析证明,该算法在可靠性、误报率、延迟和消息开销量上比同类算法具有优势.

  18. Simultaneous detection of folic acid and methotrexate by an optical sensor based on molecularly imprinted polymers on dual-color CdTe quantum dots.

    Science.gov (United States)

    Ensafi, Ali A; Nasr-Esfahani, Parisa; Rezaei, B

    2017-12-15

    In this work, molecularly imprinted polymers (MIPs) were used on the surface of cadmium telluride quantum dots (CdTe QDs) for the simultaneous determination of folic acid (FA) and methotrexate (MTX). For this purpose, two different sizes of CdTe QDs with emission peaks in the yellow (QD Y ) and orange (QD O ) spectral regions were initially synthesized and capped with MIPs. FA and MTX were used as templates for the synthesis of the two composites and designated as QD Y -MIPs and QD O -MIPs, respectively. Fourier transform infrared spectroscopy, transmission electron microscopy, and fluorescence spectroscopy were employed to characterize the composites. QD Y -MIPs and QD O -MIPs were then mixed (to form QDs-MIPs) and excited at identical excitation wavelengths; they emitted two different emission wavelengths without any spectral overlap. The fluorescence signals of QD Y -MIPs and QD O -MIPs diminished in intensity with increasing concentration of the corresponding template molecules. Under optimal conditions, the dynamic range was 0.5-20 μmol L -1 for FA and MTX, and the detection limits for FA and MTX were 32.0 nmol L -1 and 34.0 nmol L -1 , respectively. The reproducibility of the method was checked for 12.5 μmol L -1 of FA and MTX to find RSD values of 4.2% and 6.3%, respectively. Finally, the applicability of the method was checked using human blood plasma samples. Results indicated the successful application of the method as a fluorescent probe for the rapid and simultaneous detection of FA and MTX in real samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. A Simultaneous Analytical Method to Profile Non-Volatile Components with Low Polarity Elucidating Differences Between Tobacco Leaves Using Atmospheric Pressure Chemical Ionization Mass Spectrometry Detection

    Directory of Open Access Journals (Sweden)

    Ishida Naoyuki

    2016-04-01

    Full Text Available A comprehensive analytical method using liquid chromatography atmospheric pressure chemical ionization mass spectrometry detector (LC/APCI-MSD was developed to determine key non-volatile components with low polarity elucidating holistic difference among tobacco leaves. Nonaqueous reversed-phase chromatography (NARPC using organic solvent ensured simultaneous separation of various components with low polarity in tobacco resin. Application of full-scan mode to APCI-MSD hyphenated with NARPC enabled simultaneous detection of numerous intense product ions given by APCI interface. Parameters for data processing to filter, feature and align peaks were adjusted in order to strike a balance between comprehensiveness and reproducibility in analysis. 63 types of components such as solanesols, chlorophylls, phytosterols, triacylglycerols, solanachromene and others were determined on total ion chromatograms according to authentic components, wavelength spectrum and mass spectrum. The whole area of identified entities among the ones detected on total ion chromatogram reached to over 60% and major entities among those identified showed favorable linearity of determination coefficient of over 0.99. The developed method and data processing procedure were therefore considered feasible for subsequent multivariate analysis. Data matrix consisting of a number of entities was then subjected to principal component analysis (PCA and hierarchical clustering analysis. Cultivars of tobacco leaves were distributed far from each cultivar on PCA score plot and each cluster seemed to be characterized by identified non-volatile components with low polarity. While fluecured Virginia (FCV was loaded by solanachromene, phytosterol esters and triacylglycerols, free phytosterols and chlorophylls loaded Burley (BLY and Oriental (ORI respectively. Consequently the whole methodology consisting of comprehensive method and data processing procedure proved useful to determine key

  20. Simultaneous determination of superoxide and hydrogen peroxide in macrophage RAW 264.7 cell extracts by microchip electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Li, Hongmin; Li, Qingling; Wang, Xu; Xu, Kehua; Chen, Zhenzhen; Gong, Xiaocong; Liu, Xin; Tong, Lili; Tang, Bo

    2009-03-15

    A method for the first time to simultaneously determine superoxide and hydrogen peroxide in macrophage RAW 264.7 cell extracts by microchip electrophoresis with laser-induced fluorescence detection (MCE-LIF) was developed. 2-Chloro-1,3-dibenzothiazolinecyclohexene (DBZTC) and bis(p-methylbenzenesulfonyl) dichlorofluorescein (FS), two probes that can be specifically derivatized by superoxide and hydrogen peroxide, respectively, were synthesized and used. Parameters influencing the derivatization and on-chip separation were optimized. With the use of a HEPES (20 mM, pH 7.4) running buffer, a 50 mm long separation channel, and a separation voltage of 1800 V, baseline separation was achieved within 48 s for the two derivatization products, DBZTC-oxide (DBO) and 2,7-dichlorofluorescein (DCF). The linearity ranges of the method were 0.08-5.0 and 0.02-5.0 microM with detection limits (signal-to-noise ratio = 3) of 10 nM (1.36 amol) and 5.6 nM (0.76 amol) for superoxide and hydrogen peroxide, respectively. The relative standard deviations (RSDs) of migration time and peak area were less than 2.0% and 5.0%, respectively. The recoveries of the cell extract samples spiked with 1.0 microM standard solutions were 96.1% and 93.0% for superoxide and hydrogen peroxide, respectively. With the use of this method, superoxide and hydrogen peroxide in phorbol myristate acetate (PMA)-stimulated macrophage RAW 264.7 cell extracts were found to be 0.78 and 1.14 microM, respectively. The method has paved a way for simultaneously determining two or more reactive oxygen species (ROS) in a biological system with high resolution.

  1. Simultaneous determination of albumin and low-molecular-mass thiols in plasma by HPLC with UV detection.

    Science.gov (United States)

    Borowczyk, Kamila; Wyszczelska-Rokiel, Monika; Kubalczyk, Paweł; Głowacki, Rafał

    2015-02-15

    In this paper, we describe a simple and robust HPLC based method for determination of total low- and high-molecular-mass thiols, protein S-linked thiols and reduced albumin in plasma. The method is based on derivatization of analytes with 2-chloro-1-methylquinolinium tetrafluoroborate, separation and quantification by reversed-phase liquid chromatography followed by UV detection. Disulfides were converted to their thiol counterparts by reductive cleavage with tris(2-carboxyethyl)phosphine. Linearity in detector response for total thiols was observed over the range of 1-40 μmol L(-1) for Hcy and glutathione (GSH), 5-100 μmol L(-1) for Cys-Gly, 20-300 μmol L(-1) for Cys and 3.1-37.5 μmol L(-1) (0.2-2.4gL(-1)) for human serum albumin (HSA). For the protein S-bound forms these values were as follows: 0.5-30 μmol L(-1) for Hcy and GSH, 2.5-60 μmol L(-1) for Cys-Gly and 5-200 μmol L(-1) for Cys. The LOQs for total HSA, Cys, Hcy, Cys-Gly and GSH were 0.5, 0.2, 0.4, 0.3 and 0.4 μmol L(-1), respectively. The estimated validation parameters for all analytes are more than sufficient to allow the analytical method to be used for monitoring of the total and protein bound thiols as well as redox status of HSA in plasma. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Simultaneous use of solution NMR and X-ray data in REFMAC5 for joint refinement/detection of structural differences

    Energy Technology Data Exchange (ETDEWEB)

    Rinaldelli, Mauro; Ravera, Enrico; Calderone, Vito; Parigi, Giacomo [University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino (Finland) (Italy); University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino (Finland) (Italy); Murshudov, Garib N., E-mail: garib@mrc-lmb.cam.ac.uk [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH (United Kingdom); Luchinat, Claudio, E-mail: garib@mrc-lmb.cam.ac.uk [University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino (Finland) (Italy); University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino (Finland) (Italy)

    2014-04-01

    Paramagnetic NMR data (pseudocontact shifts and self-orientation residual dipolar couplings) and diamagnetic residual dipolar couplings can now be used in the program REFMAC5 from CCP4 as structural restraints together with X-ray crystallographic data. These NMR restraints can reveal differences between solid state and solution conformations of molecules or, in their absence, can be used together with X-ray crystall