WorldWideScience

Sample records for diagnostics microchips detection

  1. Nanostructured optical microchips for cancer biomarker detection.

    Science.gov (United States)

    Zhang, Tianhua; He, Yuan; Wei, Jianjun; Que, Long

    2012-01-01

    Herein we report the label-free detection of a cancer biomarker using newly developed arrayed nanostructured Fabry-Perot interferometer (FPI) microchips. Specifically, the prostate cancer biomarker free prostate-specific antigen (f-PSA) has been detected with a mouse anti-human PSA monoclonal antibody (mAb) as the receptor. Experiments found that the limit-of-detection of current nanostructured FPI microchip for f-PSA is about 10 pg/mL and the upper detection range for f-PSA can be dynamically changed by varying the amount of the PSA mAb immobilized on the sensing surface. The control experiments have also demonstrated that the immunoassay protocol used in the experiments shows excellent specificity and selectivity, suggesting the great potential to detect the cancer biomarkers at trace levels in complex biofluids. In addition, given its nature of low cost, simple-to-operation and batch fabrication capability, the arrayed nanostructured FPI microchip-based platform could provide an ideal technical tool for point-of-care diagnostics application and anticancer drug screen and discovery.

  2. Recent developments in electrochemical detection for microchip capillary electrophoresis.

    Science.gov (United States)

    Vandaveer, Walter R; Pasas-Farmer, Stephanie A; Fischer, David J; Frankenfeld, Celeste N; Lunte, Susan M

    2004-11-01

    Significant progress in the development of miniaturized microfluidic systems has occurred since their inception over a decade ago. This is primarily due to the numerous advantages of microchip analysis, including the ability to analyze minute samples, speed of analysis, reduced cost and waste, and portability. This review focuses on recent developments in integrating electrochemical (EC) detection with microchip capillary electrophoresis (CE). These detection modes include amperometry, conductimetry, and potentiometry. EC detection is ideal for use with microchip CE systems because it can be easily miniaturized with no diminution in analytical performance. Advances in microchip format, electrode material and design, decoupling of the detector from the separation field, and integration of sample preparation, separation, and detection on-chip are discussed. Microchip CEEC applications for enzyme/immunoassays, clinical and environmental assays, as well as the detection of neurotransmitters are also described.

  3. A graphene-modified cellulose paper microchip for HIV detection

    Science.gov (United States)

    Safavieh, Mohammadali; Khetani, Sultan; Kaul, Vivasvat; Kuritzkes, Daniel R.; Shafiee, Hadi

    2015-05-01

    Rapid and inexpensive virus detection and quantification at the point-of-care is of paramount importance for HIV management in resource-limited settings. Here, we report on an easy-to-fabricate, cellulose paper-based microchip with printed graphene-modified electrodes for rapid detection of HIV-1 through electrical sensing. We evaluated the effect of electrode material and geometry on the performance of the microchip to detect serially diluted, electrically conductive samples. We evaluated the optimized microchip with HIVspiked samples.

  4. New analytical portable instrument for microchip electrophoresis with electrochemical detection.

    Science.gov (United States)

    Fernández-la-Villa, Ana; Pozo-Ayuso, Diego F; Castaño-Alvarez, Mario

    2010-08-01

    A new portable instrument that includes a high voltage power supply, a bipotentiostat, and a chip holder has been especially developed for using microchips electrophoresis with electrochemical detection. The main unit of the instrument has dimensions of 150 x 165 x 70 mm (wxdxh) and consists of a four-outputs high voltage power supply with a maximum voltage of +/-3 KV and an acquisition system with two channels for dual amperometric (DC or pulsed amperometric detection) detection. Electrochemical detection has been selected as signal transduction method because it is relatively easily implemented, since nonoptical elements are required. The system uses a lithium-ion polymer battery and it is controlled from a desktop or laptop PC with a graphical user interface based on LabVIEW connected by serial RS232 or Bluetooth. The last part of the system consists of a reusable chip holder for housing the microchips, which contain all the electrical connections and reservoirs for making the work with microchips easy. The performance of the new instrument has been evaluated and compared with other commercially available apparatus using single- and dual-channel pyrex microchips for the separation of the neurotransmitters dopamine, epinephrine, and 3,4-dihydroxy-L-phenyl-alanine. The reduction of the size of the instrument has not affected the good performance of the separation and detection using microchips electrophoresis with electrochemical detection. Moreover, the new portable instrument paves the way for in situ analysis making the use of microchips electrophoresis easier.

  5. Multiplexed detection of biological agents using optical microchip sensors

    Science.gov (United States)

    Bhatta, D.; McDonnell, M. B.; Perkins, E.

    2010-10-01

    A multi-channel optical microchip sensor system suitable for real-time, label-free detection of a wide range of biological agents is presented. SpectroSensTM chips containing multiple high-precision planar Bragg gratings are exploited as lowcost, robust refractive index sensors. Sensitivity to biological agents is conferred by functionalising individual sensing regions with different antibodies selected against numerous targets of interest. Antigen binding to the surfaceimmobilised antibodies results in localised changes in refractive index; upon laser-induced interrogation of the sensing region via optical fibres, these antibody-antigen interactions manifest as increases in wavelength of light reflected from the sensor chip. Real-time detection of multiple biological agents including bacterial cells/spores, viruses and toxins has been demonstrated. Further improvements to sensor performance including physical and chemical methods are also investigated. This multi-analyte capability highlights the potential use of this sensing technology in applications ranging from bio-hazard detection for defence purposes to point-of-care clinical diagnostics.

  6. Variability of microchip capillary electrophoresis with conductivity detection.

    Science.gov (United States)

    Tantra, Ratna; Robinson, Kenneth; Sikora, Aneta

    2014-02-01

    Microfluidic CE with conductivity detection platforms could have an impact on the future development of smaller, faster and portable devices. However, for the purpose of reliable identification and quantification, there is a need to understand the degree of irreproducibility associated with the analytical technique. In this study, a protocol was developed to remove baseline drift problems sometimes observed in such devices. The protocol, which consisted of pre-conditioning steps prior to analysis, was used to further assess measurement variability from 24 individual microchips fabricated from six separate batches of glass substrate. Results show acceptable RSD percentage for retention time measurements but large variability in their corresponding peak areas (with some microchips having variability of ∼50%). Sources of variability were not related to substrate batch but possibly to a number of factors such as applied voltage fluctuations or variations in microchannel quality, for example surface roughness that will subsequently affect microchannel dimensions.

  7. Apolipoprotein B100 analysis in microchip with electrochemical detection

    Institute of Scientific and Technical Information of China (English)

    Cheng Cheng Liu; Yun Liu; Hui Xiang Wang; Yan Bo Qi; Peng Yuan Yang; Bao Hong Liu

    2011-01-01

    Apolipoprotein B100 (apoB-100) is a major protein of the cholesterol-rich low-density lipoprotein (LDL) and reflects a better assessment of total atherogenic burden to the vascular system than LDL. In this work, a simple and sensitive method has been developed to determine picoliter apoB-100s using the PMMA microfluidic chip coupled with electrochemical detection system. This method performs very well with a detectable linear range of 1-800 pg/mL and a detection limit of 1 pg/mL. A real serum sample has further been detected by this microchip-based biosensor. The results show that this kind of method is practicable and has the potential application in clinical analysis and diagnosis.

  8. Electrode substrate innovation for electrochemical detection in microchip electrophoresis.

    Science.gov (United States)

    Randviir, Edward P; Banks, Craig E

    2015-08-01

    Microchip electrophoresis (MCE) represents the next generation of miniaturised electrophoretic devices and carry benefits such as significant improvement in analysis times, lower consumption of reagents and samples, flexibility and procedural simplicity. The devices provide a separation method for complex sample matrices and an on-board detection method for the analytical determination of a target compound. The detection part of MCE is increasingly leaning towards electrochemical methods, thus the selectivity and sensitivity of detection in MCE is dependent upon the chosen working electrode composition in addition to operating conditions of the chip such as separation voltage. Given the current plethora of electrode materials that are available, there exists a possibility to creatively integrate electrodes into MCE. This review will overview the application of several electrode materials, from the old through to the new. A particular recent focus has been the selectivity element of MCEs overcome with the use of enzymes, carbon composites and screen-printed technologies.

  9. Light emitting diode, photodiode-based fluorescence detection system for DNA analysis with microchip electrophoresis.

    Science.gov (United States)

    Hall, Gordon H; Glerum, D Moira; Backhouse, Christopher J

    2016-02-01

    Electrophoretic separation of fluorescently end-labeled DNA after a PCR serves as a gold standard in genetic diagnostics. Because of their size and cost, instruments for this type of analysis have had limited market uptake, particularly for point-of-care applications. This might be changed through a higher level of system integration and lower instrument costs that can be realized through the use of LEDs for excitation and photodiodes for detection--if they provide sufficient sensitivity. Here, we demonstrate an optimized microchip electrophoresis instrument using polymeric fluidic chips with fluorescence detection of end-labeled DNA with a LOD of 0.15 nM of Alexa Fluor 532. This represents orders of magnitude improvement over previously reported instruments of this type. We demonstrate the system with an electrophoretic separation of two PCR products and their respective primers. We believe that this is the first LED-induced fluorescence microchip electrophoresis system with photodiode-based detection that could be used for standard applications of PCR and electrophoresis.

  10. PMMA-based capillary electrophoresis electrochemical detection microchip fabrication

    Science.gov (United States)

    Horng, Ray-Hua; Han, Pin; Chen, Hung-Yu; Lin, Kuan-Wen; Tsai, Tung-Mung; Zen, Jyh-Myng

    2005-01-01

    In this paper, a 50 µm (depth) × 50 µm (width) microfluidic channel is made on a poly(methyl methacrylate) (PMMA) substrate using thick photoresist. Openings were drilled for buffer reservoirs on an additional piece of PMMA. A final PMMA/patterned photoresist/PMMA sandwich configuration was completed using a bonding process. The thick photoresist was used as the adhesion layer and also as the microfluidic system. Using screen-printed technology for carbon and silver electrode fabrication, the microchip electrophoretic device functions were demonstrated. Successful detection of uric acid and L-ascorbic acid (the main components in human urine) validates the functionality of the proposed system. Successful ascorbic and uric acid separation in a sample from a urine donor who had consumed 500 mg of vitamins verified the proposed biochip.

  11. Microchip-Based Organophosphorus Detection Using Bienzyme Bioelectrocatalysis

    Science.gov (United States)

    Han, Yong Duk; Jeong, Chi Yong; Lee, Jun Hee; Lee, Dae-Sik; Yoon, Hyun C.

    2012-06-01

    We have developed a microsystem for the detection of organophosphorus (OP) compounds using acetylcholine esterase (AchE) and choline oxidase (ChOx) bienzyme bioelectrocatalysis. Because AchE is irreversibly inhibited by OP pesticides, the change in AchE activity with OP treatment can be traced to determine OP concentration. Polymer-associated ChOx immobilization on the working electrode surface and magnetic microparticle (MP)-assisted AchE deposition methods were employed to create an AchE-ChOx bienzyme-modified biosensing system. ChOx was immobilized on the micropatterned electrodes using poly(L-lysine), glutaraldehyde, and amine-rich interfacial surface. AchE was immobilized on the MP surface via Schiff's base formation, and the enzyme-modified MPs were deposited on the working electrode using a magnet under the microfluidic channel. The bioelectrocatalytic reaction between AchE-ChOx bienzyme cascade and the ferrocenyl electron shuttle was successfully used to detect OP with the developed microchip. This provides a self-contained and relatively easy method for OP detection. It requires minimal time and a small sample size, and has potential analytic applications in pesticides and chemical warfare agents.

  12. An integrated micro-chip for rapid detection of magnetic particles

    KAUST Repository

    Gooneratne, Chinthaka P.

    2012-03-09

    This paper proposes an integrated micro-chip for the manipulation and detection of magnetic particles (MPs). A conducting ring structure is used to manipulate MPs toward giant magnetoresistance(GMR) sensing elements for rapid detection. The GMRsensor is fabricated in a horseshoe shape in order to detect the majority of MPs that are trapped around the conducting structure. The GMR sensing elements are connected in a Wheatstone bridge circuit topology for optimum noise suppression. Full fabrication details of the micro-chip, characterization of the GMRsensors, and experimental results with MPs are presented in this paper. Experimental results showed that the micro-chip can detect MPs from low concentration samples after they were guided toward the GMRsensors by applying current to the conducting ring structure.

  13. Quantitative wavelength-resolved fluorescence detection for microchip capillary electrophoresis

    NARCIS (Netherlands)

    Götz, Sebastian

    2006-01-01

    This thesis describes the development and application of a new wavelengthresolved CCD-based fluorescence detector for microchip separations. In recent years, miniaturization has been one of the major trends in the development of new analytical separation systems. As the manipulated sample amounts an

  14. Construction and evaluation of a novel end-column amperometric detection system for electrophoresis microchips

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A set of integrated end-column amperometric detection system has been developed,onto which an electrophoresis microchip can be conveniently integrated.Finely machined by a piece of transparent organic glass,the system consists of an electrophoresis microchip platform and an amperometric detection reservoir,in which the microchip can be fixed onto the platform by microchip grooves and with stainless steel fixture.Each detection electrode can be directly fixed in the amperometric detection reservoir by screws and nuts.With dopamine as the model analyte,we take platinum disc electrodes with different diameters of 100 μm,300 μm and 500 μm and a carbon fiber electrode with the diameter of 240 μm as the working electrode,all of which accomplish sensitive detection.The detection parameters of the system are optimized with the carbon fiber electrode.The detection results show that in the electrochemical cleaning procedure,the relative deviations of 3.2% and 0.5% for the peak current and retention time,respectively,can be obtained for the successive detections of 100 μM dopamine,and the limit of detection for dopamine can reach 0.4 μM(S/N = 3).This system is small,but its performance of detection is stable and sensitive,and the replacement of its working electrodes is convenient,so it is very suitable as a universal platform of end-column amperometric detection for electrophoresis microchips.

  15. A microchip electrophoresis system with integrated in-plane electrodes for contactless conductivity detection

    NARCIS (Netherlands)

    Lichtenberg, Jan; de Rooij, Nico F.; Verpoorte, Elisabeth

    2002-01-01

    We present a new approach for contactless conductivity detection for microchip-based capillary electrophoresis (CE). The detector integrates easily with well-known microfabrication techniques for glass-based microfluidic devices. Platinum electrodes are structured in recesses in-plane with the micro

  16. Rapid detection of EBOLA VP40 in microchip immunofiltration assay

    Science.gov (United States)

    Miethe, Peter; Gary, Dominik; Hlawatsch, Nadine; Gad, Anne-Marie

    2015-05-01

    In the spring of 2014, the Ebola virus (EBOV) strain Zaire caused a dramatic outbreak in several regions of West Africa. The RT-PCR and antigen capture diagnostic proved to be effective for detecting EBOV in blood and serum. In this paper, we present data of a rapid antigen capture test for the detection of VP40. The test was performed in a microfluidic chip for immunofiltration analysis. The chip integrates all necessary assay components. The analytical sensitivity of the rapid test was 8 ng/ml for recombinant VP40. In serum and whole blood samples spiked with virus culture material, the detection limit was 2.2 x 102 PFU/ml. The performance data of the rapid test (15 min) are comparable to that of the VP40 laboratory ELISA.

  17. Inner structure detection by optical tomography technology based on feedback of microchip Nd:YAG lasers.

    Science.gov (United States)

    Xu, Chunxin; Zhang, Shulian; Tan, Yidong; Zhao, Shijie

    2013-05-20

    We describe a new optical tomography technology based on feedback of microchip Nd:YAG lasers. In the case of feedback light frequency-shifted, light can be magnified by a fact of 10(6) in the Nd:YAG microchip lasers, which makes it possible to realize optical tomography with a greater depth than current optical tomography. The results of the measuring and imaging of kinds of samples are presented, which demonstrate the feasibility and potential of this approach in the inner structure detection. The system has a lateral resolution of ~1 μm, a vertical resolution of 15 μm and a longitudinal scanning range of over 10mm.

  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

    is collected by another waveguide in the opposite end of the detection cell, while visible fluorescence is collected vertically through the lid of the chip. The background of scattered excitation light is suppressed by detection perpendicular to the excitation, the limited UV transparency of the borosilicate......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...... compounds could be detected in the micromolar range by absorbance detection, but fluorescence detection improved detection limits for compounds displaying native UV fluorescence up to ten times. The simultaneous detection also proved useful for the identification of compounds with similar retention times...

  19. Side-by-side comparison of disposable microchips with commercial capillary cartridges for application in capillary isoelectric focusing with whole column imaging detection.

    Science.gov (United States)

    Liu, Zhen; Ou, Junjie; Samy, Razim; Glawdel, Tomasz; Huang, Tiemin; Ren, Carolyn L; Pawliszyn, Janusz

    2008-10-01

    Simple-structured, well-functioned disposable poly(dimethylsiloxane) (PDMS) microchips were developed for capillary isoelectric focusing with whole column imaging detection (CIEF-WCID). Side-by-side comparison of the developed microchips with well-established commercial capillary cartridges demonstrated that the disposable microchips have comparable performance as well as advantages such as absence of lens effect and possibility of high-aspect-ratio accompanied with a dramatic reduction in cost.

  20. Use of epoxy-embedded electrodes to integrate electrochemical detection with microchip-based analysis systems.

    Science.gov (United States)

    Selimovic, Asmira; Johnson, Alicia S; Kiss, István Z; Martin, R Scott

    2011-04-01

    A new method of fabricating electrodes for microchip devices that involves the use of Teflon molds and a commercially available epoxy to embed electrodes of various sizes and compositions is described. The resulting epoxy base can be polished to generate a fresh electrode and sealed against poly(dimethylsiloxane) (PDMS)-based fluidic structures. Microchip-based flow injection analysis was used to characterize the epoxy-embedded electrodes. It was shown that gold electrodes can be amalgamated with liquid mercury and the resulting mercury/gold electrode is used to selectively detect glutathione from lysed red blood cells. The ability to encapsulate multiple electrode materials of differing compositions enabled the integration of microchip electrophoresis with electrochemical detection. Finally, a unique feature of this approach is that the electrode connection is made from the bottom of the epoxy base. This enables the creation of three-dimensional gold pillar electrodes (65 μm in diameter and 27 μm in height) that can be integrated within a fluidic network. As compared with the use of a flat electrode of a similar diameter, the use of the pillar electrode led to improvements in both the sensitivity (72.1 pA/μM for the pillar versus 4.2 pA/μM for the flat electrode) and limit of detection (20 nM for the pillar versus 600 nM for the flat electrode), with catechol being the test analyte. These epoxy-embedded electrodes hold promise for the creation of inexpensive microfluidic devices that can be used to electrochemically detect biologically important analytes in a manner where the electrodes can be polished and a fresh electrode surface is generated as desired.

  1. Nonlinear multi-photon laser wave-mixing optical detection in microarrays and microchips for ultrasensitive detection and separation of biomarkers for cancer and neurodegenerative diseases

    Science.gov (United States)

    Iwabuchi, Manna; Hetu, Marcel; Maxwell, Eric; Pradel, Jean S.; Ramos, Sashary; Tong, William G.

    2015-09-01

    Multi-photon degenerate four-wave mixing is demonstrated as an ultrasensitive absorption-based optical method for detection, separation and identification of biomarker proteins in the development of early diagnostic methods for HIV- 1, cancer and neurodegenerative diseases using compact, portable microarrays and capillary- or microchip-based chemical separation systems that offer high chemical specificity levels. The wave-mixing signal has a quadratic dependence on concentration, and hence, it allows more reliable monitoring of smaller changes in analyte properties. Our wave-mixing detection sensitivity is comparable or better than those of current methods including enzyme-linked immunoassay for clinical diagnostic and screening. Detection sensitivity is excellent since the wave-mixing signal is a coherent laser-like beam that can be collected with virtually 100% collection efficiency with high S/N. Our analysis time is short (1-15 minutes) for molecular weight-based protein separation as compared to that of a conventional separation technique, e.g., sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When ultrasensitive wavemixing detection is paired with high-resolution capillary- or microchip-based separation systems, biomarkers can be separated and identified at the zepto- and yocto-mole levels for a wide range of analytes. Specific analytes can be captured in a microchannel through the use of antibody-antigen interactions that provide better chemical specificity as compared to size-based separation alone. The technique can also be combined with immune-precipitation and a multichannel capillary array for high-throughput analysis of more complex protein samples. Wave mixing allows the use of chromophores and absorption-modifying tags, in addition to conventional fluorophores, for online detection of immunecomplexes related to cancer.

  2. Capillary electrophoresis-electrochemical detection microchip device and supporting circuits

    Science.gov (United States)

    Jackson, Douglas J.; Roussel, Jr., Thomas J.; Crain, Mark M.; Baldwin, Richard P.; Keynton, Robert S.; Naber, John F.; Walsh, Kevin M.; Edelen, John. G.

    2008-03-18

    The present invention is a capillary electrophoresis device, comprising a substrate; a first channel in the substrate, and having a buffer arm and a detection arm; a second channel in the substrate intersecting the first channel, and having a sample arm and a waste arm; a buffer reservoir in fluid communication with the buffer arm; a waste reservoir in fluid communication with the waste arm; a sample reservoir in fluid communication with the sample arm; and a detection reservoir in fluid communication with the detection arm. The detection arm and the buffer arm are of substantially equal length.

  3. Rapid and sensitive measurements of nitrate ester explosives using microchip electrophoresis with electrochemical detection.

    Science.gov (United States)

    Piccin, Evandro; Dossi, Nicolò; Cagan, Avi; Carrilho, Emanuel; Wang, Joseph

    2009-03-01

    This article describes an effective microchip protocol based on electrophoretic-separation and electrochemical detection for highly sensitive and rapid measurements of nitrate ester explosives, including ethylene glycol dinitrate (EGDN), pentaerythritol tetranitrate (PETN), propylene glycol dinitrate (PGDN) and glyceryl trinitrate (nitroglycerin, NG). Factors influencing the separation and detection processes were examined and optimized. Under the optimal separation conditions obtained using a 15 mM borate buffer (pH 9.2) containing 20 mM SDS, and applying a separation voltage of 1500 V, the four nitrate ester explosives were separated within less than 3 min. The glassy-carbon amperometric detector (operated at -0.9 V vs. Ag/AgCl) offers convenient cathodic detection down to the picogram level, with detection limits of 0.5 ppm and 0.3 ppm for PGDN and for NG, respectively, along with good repeatability (RSD of 1.8-2.3%; n = 6) and linearity (over the 10-60 ppm range). Such effective microchip operation offers great promise for field screening of nitrate ester explosives and for supporting various counter-terrorism surveillance activities.

  4. Poly(dimethylsiloxane) microchip-based immunoassay with multiple reaction zones: Toward on-chip multiplex detection platform

    Energy Technology Data Exchange (ETDEWEB)

    Shao, Guocheng; Wang, Jun; Li, Zhaohui; Saraf, Laxmikant V.; Wang, Wanjun; Lin, Yuehe

    2011-09-20

    In this work, a poly(dimethylsiloxane) (PDMS) microchip-based immuno-sensing platform with integrated pneumatic micro valves is described. The microchip was fabricated with multiple layer soft lithography technology. By controlling the activation status of corresponding valves, reagent flows in the microchannel network can be well manipulated so that immuno-reactions only take place at designated reaction zones (DRZs). Four DRZs are included in the prototype microchip. Since these DRZs are all isolated from each other by micro valves, cross contamination is prevented. Using the inner surface of the all-PDMS microchannel as immunoassay substrate, on-chip sandwich format solid phase immunoassay was performed to demonstrate the feasibility of this immuno-sensing platform. Mouse IgG and fluorescein isothiocyanate (FITC) were used as the model analyte and the signal reporter respectively. Only 10 ul sample is needed for the assay and low detection limit of 5 ng/ml (≈33 pM) was achieved though low-cost polyclonal antibodies were used in our experiment for feasibility study only. The encouraging results from mouse IgG immunoassay proved the feasibility of our microchip design. With slight modification of the assay protocol, the same chip design can be used for multi-target detection and can provide a simple, cost-effective and integrated microchip solution for multiplex immunoassay applications.

  5. Integrated hybrid polystyrene-polydimethylsiloxane device for monitoring cellular release with microchip electrophoresis and electrochemical detection

    Science.gov (United States)

    Johnson, Alicia S.; Mehl, Benjamin T.; Martin, R. Scott

    2015-01-01

    In this work, a polystyrene (PS)-polydimethylsiloxane (PDMS) hybrid device was developed to enable the integration of cell culture with analysis by microchip electrophoresis and electrochemical detection. It is shown that this approach combines the fundamental advantages of PDMS devices (the ability to integrate pumps and valves) and PS devices (the ability to permanently embed fluidic tubing and electrodes). The embedded fused-silica capillary enables high temporal resolution measurements from off-chip cell culture dishes and the embedded electrodes provide close to real-time analysis of small molecule neurotransmitters. A novel surface treatment for improved (reversible) adhesion between PS and PDMS is described using a chlorotrimethylsilane stamping method. It is demonstrated that a Pd decoupler is efficient at handling the high current (and cathodic hydrogen production) resulting from use of high ionic strength buffers needed for cellular analysis; thus allowing an electrophoretic separation and in-channel detection. The separation of norepinephrine (NE) and dopamine (DA) in highly conductive biological buffers was optimized using a mixed surfactant system. This PS-PDMS hybrid device integrates multiple processes including continuous sampling from a cell culture dish, on-chip pump and valving technologies, microchip electrophoresis, and electrochemical detection to monitor neurotransmitter release from PC 12 cells. PMID:25663849

  6. Bio-inspired photonic-crystal microchip for fluorescent ultratrace detection.

    Science.gov (United States)

    Hou, Jue; Zhang, Huacheng; Yang, Qiang; Li, Mingzhu; Song, Yanlin; Jiang, Lei

    2014-06-02

    Ultratrace detection attracts great interest because it is still a challenge to the early diagnosis and drug testing. Enriching the targets from highly diluted solutions to the sensitive area is a promising method. Inspired by the fog-collecting structure on Stenocara beetle's back, a photonic-crystal (PC) microchip with hydrophilic-hydrophobic micropattern was fabricated by inkjet printing. This device was used to realize high-sensitive ultratrace detection of fluorescence analytes and fluorophore-based assays. Coupled with the fluorescence enhancement effect of a PC, detection down to 10(-16) mol L(-1) was achieved. This design can be combined with biophotonic devices for the detection of drugs, diseases, and pollutions of the ecosystem.

  7. A Microchip for Integrated Single-Cell Gene Expression Profiling and Genotoxicity Detection

    Directory of Open Access Journals (Sweden)

    Hui Dong

    2016-09-01

    Full Text Available Microfluidics-based single-cell study is an emerging approach in personalized treatment or precision medicine studies. Single-cell gene expression holds a potential to provide treatment selections with maximized efficacy to help cancer patients based on a genetic understanding of their disease. This work presents a multi-layer microchip for single-cell multiplexed gene expression profiling and genotoxicity detection. Treated by three drug reagents (i.e., methyl methanesulfonate, docetaxel and colchicine with varied concentrations and time lengths, individual human cancer cells (MDA-MB-231 are lysed on-chip, and the released mRNA templates are captured and reversely transcribed into single strand DNA. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, cyclin-dependent kinase inhibitor 1A (CDKN1A, and aurora kinase A (AURKA genes from single cells are amplified and real-time quantified through multiplex polymerase chain reaction. The microchip is capable of integrating all steps of single-cell multiplexed gene expression profiling, and providing precision detection of drug induced genotoxic stress. Throughput has been set to be 18, and can be further increased following the same approach. Numerical simulation of on-chip single cell trapping and heat transfer has been employed to evaluate the chip design and operation.

  8. Multicolor instrumentation for direct fluorescent detection of nucleic acids in a microchip format

    Science.gov (United States)

    Bogdanov, Valery L.; Rogers, Yu-Hui; Lan, Guang; Boyce-Jacino, Michael

    1998-04-01

    Deposition of nucleic acids on solid support in the form of high density arrays (a DNA microarray) creates a powerful nonelectrophoretic technology for highly parallel genetic analysis. Microarrays have applications in the areas of DNA sequencing, genetic mutation detection and gene expression monitoring. We report here the design and utility of an experimental instrument for microchip parallel hyperspectral fluorescent imaging. The instrument integrates in-line laser excitation of microarray, parallel fluorescent spectrometry with cooled CCD and dye-base spectral classification software. Instrument has been applied for imaging detection, spectral analysis and base classification of Genetic Bit Analysis (GBA) reactions in a microchip format on a glass support. GBA is a solid phase DNA sequence analysis method that provides single nucleotide resolution by specific extension of dye-labeled dideoxynucleotidetriphosphates (ddNTPs). GBA testing yields one or two different ddNTPs on any given microarray spot, so analysis must resolve any pair wise combination of all possible ddNTPs labeled with distinct fluorescent dyes.

  9. A Microchip for Integrated Single-Cell Gene Expression Profiling and Genotoxicity Detection

    Science.gov (United States)

    Dong, Hui; Sun, Hao

    2016-01-01

    Microfluidics-based single-cell study is an emerging approach in personalized treatment or precision medicine studies. Single-cell gene expression holds a potential to provide treatment selections with maximized efficacy to help cancer patients based on a genetic understanding of their disease. This work presents a multi-layer microchip for single-cell multiplexed gene expression profiling and genotoxicity detection. Treated by three drug reagents (i.e., methyl methanesulfonate, docetaxel and colchicine) with varied concentrations and time lengths, individual human cancer cells (MDA-MB-231) are lysed on-chip, and the released mRNA templates are captured and reversely transcribed into single strand DNA. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cyclin-dependent kinase inhibitor 1A (CDKN1A), and aurora kinase A (AURKA) genes from single cells are amplified and real-time quantified through multiplex polymerase chain reaction. The microchip is capable of integrating all steps of single-cell multiplexed gene expression profiling, and providing precision detection of drug induced genotoxic stress. Throughput has been set to be 18, and can be further increased following the same approach. Numerical simulation of on-chip single cell trapping and heat transfer has been employed to evaluate the chip design and operation. PMID:27649175

  10. Barcoded microchips for biomolecular assays.

    Science.gov (United States)

    Zhang, Yi; Sun, Jiashu; Zou, Yu; Chen, Wenwen; Zhang, Wei; Xi, Jianzhong Jeff; Jiang, Xingyu

    2015-01-20

    Multiplexed assay of analytes is of great importance for clinical diagnostics and other analytical applications. Barcode-based bioassays with the ability to encode and decode may realize this goal in a straightforward and consistent manner. We present here a microfluidic barcoded chip containing several sets of microchannels with different widths, imitating the commonly used barcode. A single barcoded microchip can carry out tens of individual protein/nucleic acid assays (encode) and immediately yield all assay results by a portable barcode reader or a smartphone (decode). The applicability of a barcoded microchip is demonstrated by human immunodeficiency virus (HIV) immunoassays for simultaneous detection of three targets (anti-gp41 antibody, anti-gp120 antibody, and anti-gp36 antibody) from six human serum samples. We can also determine seven pathogen-specific oligonucleotides by a single chip containing both positive and negative controls.

  11. Fully automated microchip system for the detection of quantal exocytosis from single and small ensembles of cells

    DEFF Research Database (Denmark)

    Spégel, Christer; Heiskanen, Arto; Pedersen, Simon

    2008-01-01

    A lab-on-a-chip device that enables positioning of single or small ensembles of cells on an aperture in close proximity to a mercaptopropionic acid (MPA) modified sensing electrode has been developed and characterized. The microchip was used for the detection of Ca2+-dependent quantal catecholami...

  12. Sensitive detection of influenza viruses with Europium nanoparticles on an epoxy silica sol-gel functionalized polycarbonate-polydimethylsiloxane hybrid microchip.

    Science.gov (United States)

    Liu, Jikun; Zhao, Jiangqin; Petrochenko, Peter; Zheng, Jiwen; Hewlett, Indira

    2016-12-15

    In an effort to develop new tools for diagnosing influenza in resource-limited settings, we fabricated a polycarbonate (PC)-polydimethylsiloxane (PDMS) hybrid microchip using a simple epoxy silica sol-gel coating/bonding method and employed it in sensitive detection of influenza virus with Europium nanoparticles (EuNPs). The incorporation of sol-gel material in device fabrication provided functionalized channel surfaces ready for covalent immobilization of primary antibodies and a strong bonding between PDMS substrates and PC supports without increasing background fluorescence. In microchip EuNP immunoassay (µENIA) of inactivated influenza viruses, replacing native PDMS microchips with hybrid microchips allowed the achievement of a 6-fold increase in signal-to-background ratio, a 12-fold and a 6-fold decreases in limit-of-detection (LOD) in influenza A and B tests respectively. Using influenza A samples with known titers, the LOD of influenza µENIA on hybrid microchips was determined to be ~10(4) TCID50 titer/mL and 10(3)-10(4) EID50 titer/mL. A comparison test indicated that the sensitivity of influenza µENIA enhanced using the hybrid microchips even surpassed that of a commercial laboratory influenza ELISA test. In addition to the sensitivity improvement, assay variation was clearly reduced when hybrid microchips instead of native PDMS microchips were used in the µENIA tests. Finally, infectious reference viruses and nasopharyngeal swab patient specimens were successfully tested using μENIA on hybrid microchip platforms, demonstrating the potential of this unique microchip nanoparticle assay in clinical diagnosis of influenza. Meanwhile, the tests showed the necessity of using nucleic acid confirmatory tests to clarify ambiguous test results obtained from prototype or developed point-of-care testing devices for influenza diagnosis.

  13. Authenticity screening of seized whiskey samples using electrophoresis microchips coupled with contactless conductivity detection.

    Science.gov (United States)

    Rezende, Kariolanda C A; Moreira, Roger Cardoso; Logrado, Lucio Paulo Lima; Talhavini, Márcio; Coltro, Wendell K T

    2016-10-01

    This report describes for the first time the use of microchip electrophoresis (ME) devices integrated with capacitively coupled contactless conductivity detection (C(4) D) to investigate the authenticity of seized whiskey samples, which were probably adulterated by simple dilution with tap water. The proposed microfluidic platform was explored for the monitoring of anionic species (Cl(-) and F(-) ) in both original and tampered samples. The best separations were achieved within 70 s using a running buffer composed of lactic acid and histidine (pH = 5.9). ME-C(4) D devices were used to analyze samples from three different brands (five samples each). Based on the presence of inorganic anions like Cl(-) , F(-) , SO4(2-) and NO2(-) in different amounts, the authenticity of seized whiskeys was compared to original samples. According to the reported data, the proposed microfluidic platform can be useful to help regulatory authorities in the investigation and monitoring of authenticity of commercialized whiskey beverages.

  14. Determination of ammonium on an integrated microchip with LED-induced fluorescence detection

    Institute of Scientific and Technical Information of China (English)

    Shuhua Xue; Katsumi Uchiyama; Hai-fang Li

    2012-01-01

    A simply fabricated microfluidic device integrated with a fluorescence detection system has been developed for on-line determination of ammonium in aqueous samples.A 365-nm light-emitting diode(LED)as an excitation source and a minor band pass filter were mounted into a polydimethylsiloxane(PDMS)-based microchip for the purpose of miniaturization of the entire analytical system.The ammonium sample reacted with o-phthaldialdehyde(OPA)on-chip with sodium sulfite as reducing reagent to produce a fluorescent isoindole derivative,which can emit fluorescence signal at about 425 nm when excited at 365 nm.Effects of pH,flow rate of solutions,concentrations of OPA-reagent,phosphate and sulfite salt were investigated.The calibration curve of ammonium in the range of 0.018-1.8 μg/mL showed a good linear relationship with R2 =0.9985,and the detection limit was(S/N =3)3.6 × 10-4 μg/mL.The relative standard deviation was 2.8%(n =11)by calculating at 0.18 μg/mL ammonium for repeated detection.The system was applied to determine the ammonium concentration in rain and river waters,even extent to other analytes fluorescence detection by the presented device.

  15. A novel capillary electrophoresis microchip with amperometric detection using a Prussian blue-modified indium tin oxide electrode

    Science.gov (United States)

    Kim, Ju-Ho; Kang, C. J.; Kim, Yong-Sang

    2005-03-01

    A novel approach to construct a disposable capillary electrophoresis microchip is proposed. The electrocatalytic oxidation of dopamine at a Prussian blue (PB)-modified indium tin oxide (ITO) electrode was described and the amperometric detection of dopamine was then investigated. The PB film on ITO electrode was electrodeposited using FeCl3 and K3Fe(CN)6 mixed solution. Our results indicated that PB film was uniform, smooth, and defect-free. The CE-chip has been tested successfully by detecting dopamine and catechol within a very short time of around 80 sec using an electric field of 60 V/cm. The results also showed that dopamine and catechol mixtures were separated efficiently and rapidly. The microsystems gave a very good reproducibility for peak height and separation time. This microchip is cost effective and adequate for a disposable sensor.

  16. [IDENTIFICATION OF A NEW DIAGNOSTIC MARKERS OF PROSTATIC CANCER, USING NOTI-MICROCHIPS].

    Science.gov (United States)

    Vozianov, S O; Kashuba, V I; Grygorenko, V M; Gordiyuk, V V; Danylets, R O; Bondarenko, Yu M; Vikarchuk, M V

    2016-04-01

    The biopsy material specimens were investigated in 33 patients, examined for the prostatic cancer suspicion. In accordance to the morphological investigation data, in 15 patients a benign prostatic hyperplasia was verified, and in 18--pancreatic adenocarcinoma. NotI-Microchips of 180 clones of the third chromosome were used for determination of epigenetic changes. In 50 genes of the third chromosome a high rate of the methylation state changes (from 33 to 82%) was noted. Some changed genes take part in cancerogenesis (HMGB1L5, LRRC58, GPR149, DZIP1L, C3orf77, NUDT16) and in the prostatic gland cancer occurrence (BCL6, ITGA9, FBLN2, SOX2, LRRC3B etc.). Dependence of the genes methylation state from the clinic-morphological indices in patients with the prostatic gland cancer, including, the prostate-specific antigen level, the tumor differentiation degree in accordance to Gleason, was not established. Panel, consisting of 16 new potential markers for early and differentiated diagnosis of prostatic gland cancer, was identified: BHLHE40, FOXP1, LOC285205, ITGA9, CTDSPL, FGF12, LOC440944/SETD5, VHL, CLCN2, OSBPL10/ZNF860, LMCD1, FAM19A4, CAND2, MAP4, KY and LRRC58.

  17. Metalless electrodes for capacitively coupled contactless conductivity detection on electrophoresis microchips.

    Science.gov (United States)

    Duarte Junior, Gerson F; Fracassi da Silva, José Alberto; Mendonça Francisco, Kelliton José; do Lago, Claudimir Lucio; Carrilho, Emanuel; Coltro, Wendell K T

    2015-08-01

    This paper describes the use of ionic solutions as sensing electrodes for capacitively coupled contactless conductivity detection on electrophoresis microchips. Initially, two channels were engraved in a PMMA holder by using a CO2 laser system and sealed with a thin adhesive membrane. PDMS electrophoresis chips were fabricated by soft lithography and reversibly sealed against the polymer membrane. Different ionic solutions were investigated as metalless electrodes. The electrode channels were filled with KCl solutions prepared in conductivity values from approximately 10 to 40 S/m. The best analytical response was achieved using the KCl solution with 21.9 S/m conductivity (2 mol/L). Besides KCl, we also tested NaCl and LiCl solutions for actuating as detection electrodes. Taking into account the same electrolyte concentration (2 mol/L), the best response was recorded with KCl solution due to its higher ionic conductivity. The optimum operating frequency (400 kHz) and the best sensing electrode (2 mol/L KCl) were used to monitor electrophoretic separations of a mixture containing K(+) , Na(+) , and Li(+) . The use of liquid solutions as sensing electrodes for capacitively coupled contactless conductivity detection measurements has revealed great performance to monitor separations on chip-based devices, avoiding complicated fabrication schemes to include metal deposition and encapsulation of electrodes. The LOD values were estimated to be 28, 40, and 58 μmol/L for K(+) , Na(+) , and Li(+) , respectively, what is comparable to that of conventional metal electrodes. When compared to the use metal electrodes, the proposed approach offers advantages regarding the easiness of fabrication, simplicity, and lower cost per device.

  18. A CCD-based fluorescence imaging system for real-time loop-mediated isothermal amplification-based rapid and sensitive detection of waterborne pathogens on microchips.

    Science.gov (United States)

    Ahmad, Farhan; Seyrig, Gregoire; Tourlousse, Dieter M; Stedtfeld, Robert D; Tiedje, James M; Hashsham, Syed A

    2011-10-01

    Rapid, sensitive, and low-cost pathogen diagnostic systems are needed for early disease diagnosis and treatment, especially in resource-limited settings. This study reports a low-cost charge-coupled device (CCD)-based fluorescence imaging system for rapid detection of waterborne pathogens by isothermal gene amplification in disposable microchips. Fluorescence imaging capability of this monochromatic CCD camera is evaluated by optimizing the gain, offset, and exposure time. This imaging system is validated for 12 virulence genes of major waterborne pathogens on cyclic olefin polymer (COP) microchips, using SYTO-82 dye and real time fluorescence loop-mediated isothermal amplification referred here as microRT(f)-LAMP. Signal-to-noise ratio (SNR) and threshold time (Tt) of microRT(f)-LAMP assays are compared with those from a commercial real-time polymerase chain reaction (PCR) instrument. Applying a CCD exposure of 5 s to 10(5) starting DNA copies of microRT(f)-LAMP assays increases the SNR by 8-fold and reduces the Tt by 9.8 min in comparison to a commercial real-time PCR instrument. Additionally, single copy level sensitivity for Campylobacter jejuni 0414 gene is obtained for microRT(f)-LAMP with a Tt of 19 min, which is half the time of the commercial real-time PCR instrument. Due to the control over the exposure time and the wide field imaging capability of CCD, this low-cost fluorescence imaging system has the potential for rapid and parallel detection of pathogenic microorganisms in high throughput microfluidic chips.

  19. Integration of a Graphite/PMMA CompositeElectrode into a Poly(methyl methacrylate) (PMMA) Substrate for Electrochemical Detection in Microchips

    Science.gov (United States)

    Regel, Anne; Lunte, Susan

    2013-01-01

    Traditional fabrication methods for polymer microchips, the bonding of two substrates together to form the microchip, can make the integration of carbon electrodes difficult. We have developed a simple and inexpensive method to integrate graphite/PMMA composite electrodes (GPCEs) into a PMMA substrate. These substrates can be bonded to other PMMA layers using a solvent-assisted thermal bonding method. The optimal composition of the GPCEs for electrochemical detection was determined using cyclic voltammetry with dopamine as a test analyte. Using the optimized GPCEs in an all-PMMA flow cell with flow injection analysis, it was possible to detect 50 nM dopamine under the best conditions. These electrodes were also evaluated for the detection of dopamine and catechol following separation by microchip electrophoresis (ME). PMID:23670816

  20. Emerging Loop-Mediated Isothermal Amplification-Based Microchip and Microdevice Technologies for Nucleic Acid Detection.

    Science.gov (United States)

    Safavieh, Mohammadali; Kanakasabapathy, Manoj K; Tarlan, Farhang; Ahmed, Minhaz U; Zourob, Mohammed; Asghar, Waseem; Shafiee, Hadi

    2016-03-14

    Rapid, sensitive, and selective pathogen detection is of paramount importance in infectious disease diagnosis and treatment monitoring. Currently available diagnostic assays based on polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) are time-consuming, complex, and relatively expensive, thus limiting their utility in resource-limited settings. Loop-mediated isothermal amplification (LAMP) technique has been used extensively in the development of rapid and sensitive diagnostic assays for pathogen detection and nucleic acid analysis and hold great promise for revolutionizing point-of-care molecular diagnostics. Here, we review novel LAMP-based lab-on-a-chip (LOC) diagnostic assays developed for pathogen detection over the past several years. We review various LOC platforms based on their design strategies for pathogen detection and discuss LAMP-based platforms still in development and already in the commercial pipeline. This review is intended as a guide to the use of LAMP techniques in LOC platforms for molecular diagnostics and genomic amplifications.

  1. Rapid separation of strychnine and brucine on a dynamically modified poly(dimethylsiloxane) microchip followed by electrochemical detection.

    Science.gov (United States)

    Zhang, Q L; Xu, J J; Lian, H Z; Li, X Y; Chen, H Y

    2006-01-01

    A method has been developed for rapidly separating and detecting strychnine and brucine using a poly(dimethysiloxane) (PDMS) microchip and electrochemical (EC) detection. PDMS microchannels dynamically modified by Brij35 are shown to be more efficient than native ones. The two analytes are well separated within 90 s in 70 mmol/L acetate buffer (pH 5.5) containing 0.01% (v/v) Brij35. Detection limits were found to be 1.0 micromol/L for strychnine and 0.2 micromol/L for brucine at S/N = 3. The method was used to determine trace strychnine and brucine in rat serum, and the results obtained correlate well with those obtained via high-performance liquid chromatography (HPLC).

  2. Hand-held analyser based on microchip electrophoresis with contactless conductivity detection for measurement of chemical warfare agent degradation products

    Science.gov (United States)

    Duran, Karolina-Petkovic; Zhu, Yonggang; Chen, Chuanpin; Swallow, Anthony; Stewart, Robert; Hoobin, Pam; Leech, Patrick; Ovenden, Simon

    2008-12-01

    This paper reports on the development of a hand-held device for on-site detection of organophosphonate nerve agent degradation products. This field-deployable analyzer relies on efficient microchip electrophoresis separation of alkyl methylphosphonic acids and their sensitive contactless conductivity detection. Miniaturized, low-powered design is coupled with promising analytical performance for separating the breakdown products of chemical warfare agents such as Soman, Sarin and VX . The detector has a detection limit of about 10 μg/mL and has a good linear response in the range 10-300 μg/mL concentration range. Applicability to environmental samples is demonstrated .The new hand-held analyzer offers great promise for converting conventional ion chromatography or capillary electrophoresis sophisticated systems into a portable forensic laboratory for faster, simpler and more reliable on-site screening.

  3. A high-performance polycarbonate electrophoresis microchip with integrated three-electrode system for end-channel amperometric detection.

    Science.gov (United States)

    Wang, Yurong; Chen, Hengwu; He, Qiaohong; Soper, Steven A

    2008-05-01

    A fully integrated polycarbonate (PC) microchip for CE with end-channel electrochemical detection operated in an amperometric mode (CE-ED) has been developed. The on-chip integrated three-electrode system consisted of a gold working electrode, an Ag/AgCl reference electrode and a platinum counter electrode, which was fabricated by photo-directed electroless plating combined with electroplating. The working electrode was positioned against the separation channel exit to reduce post-channel band broadening. The electrophoresis high-voltage (HV) interference with the amperometric detection was assessed with respect to detection noise and potential shifts at various working-to-reference electrode spacing. It was observed that the electrophoresis HV interference caused by positioning the working electrode against the channel exit could be diminished by using an on-chip integrated reference electrode that was positioned in close proximity (100 microm) to the working electrode. The CE-ED microchip was demonstrated for the separation of model analytes, including dopamine (DA) and catechol (CA). Detection limits of 132 and 164 nM were achieved for DA and CA, respectively, and a theoretical plate number of 2.5x10(4)/m was obtained for DA. Relative standard deviations in peak heights observed for five runs of a standard solution containing the two analytes (0.1 mM for each) were 1.2 and 3.1% for DA and CA, respectively. The chip could be continuously used for more than 8 h without significant deterioration in analytical performance.

  4. Integration of a graphite/poly(methyl-methacrylate) composite electrode into a poly(methylmethacrylate) substrate for electrochemical detection in microchips.

    Science.gov (United States)

    Regel, Anne; Lunte, Susan

    2013-07-01

    Traditional fabrication methods for polymer microchips, the bonding of two substrates together to form the microchip, can make the integration of carbon electrodes difficult. We have developed a simple and inexpensive method to integrate graphite/PMMA composite electrodes (GPCEs) into a PMMA substrate. These substrates can be bonded to other PMMA layers using a solvent-assisted thermal bonding method. The optimal composition of the GPCEs for electrochemical detection was determined using cyclic voltammetry with dopamine as a test analyte. Using the optimized GPCEs in an all-PMMA flow cell with flow injection analysis, it was possible to detect 50 nM dopamine under the best conditions. These electrodes were also evaluated for the detection of dopamine and catechol following separation by MCE.

  5. Simultaneous detection of 19 K-ras mutations by free-solution conjugate electrophoresis of ligase detection reaction products on glass microchips.

    Science.gov (United States)

    Albrecht, Jennifer Coyne; Kotani, Akira; Lin, Jennifer S; Soper, Steven A; Barron, Annelise E

    2013-02-01

    We demonstrate here the power and flexibility of free-solution conjugate electrophoresis (FSCE) as a method of separating DNA fragments by electrophoresis with no sieving polymer network. Previous work introduced the coupling of FSCE with ligase detection reaction (LDR) to detect point mutations, even at low abundance compared to the wild-type DNA. Here, four large drag-tags are used to achieve free-solution electrophoretic separation of 19 LDR products ranging in size from 42 to 66 nt that correspond to mutations in the K-ras oncogene. LDR-FSCE enabled electrophoretic resolution of these 19 LDR-FSCE products by CE in 13.5 min (E = 310 V/cm) and by microchip electrophoresis in 140 s (E = 350 V/cm). The power of FSCE is demonstrated in the unique characteristic of free-solution separations where the separation resolution is constant no matter the electric field strength. By microchip electrophoresis, the electric field was increased to the maximum of the power supply (E = 700 V/cm), and the 19 LDR-FSCE products were separated in less than 70 s with almost identical resolution to the separation at E = 350 V/cm. These results will aid the goal of screening K-ras mutations on integrated "sample-in/answer-out" devices with amplification, LDR, and detection all on one platform.

  6. High-sensitivity microchip electrophoresis determination of inorganic anions and oxalate in atmospheric aerosols with adjustable selectivity and conductivity detection.

    Science.gov (United States)

    Noblitt, Scott D; Schwandner, Florian M; Hering, Susanne V; Collett, Jeffrey L; Henry, Charles S

    2009-02-27

    A sensitive and selective separation of common anionic constituents of atmospheric aerosols, sulfate, nitrate, chloride, and oxalate, is presented using microchip electrophoresis. The optimized separation is achieved in under 1 min and at low background electrolyte ionic strength (2.9 mM) by combining a metal-binding electrolyte anion (17 mM picolinic acid), a sulfate-binding electrolyte cation (19 mM HEPBS), a zwitterionic surfactant with affinity towards weakly solvated anions (19 mM N-tetradecyl,N,N-dimethyl-3-ammonio-1-propansulfonate), and operation in counter-electroosmotic flow (EOF) mode. The separation is performed at pH 4.7, permitting pH manipulation of oxalate's mobility. The majority of low-concentration organic acids are not observed at these conditions, allowing for rapid subsequent injections without the presence of interfering peaks. Because the mobilities of sulfate, nitrate, and oxalate are independently controlled, other minor constituents of aerosols can be analyzed, including nitrite, fluoride, and formate if desired using similar separation conditions. Contact conductivity detection is utilized, and the limit of detection for oxalate (S/N=3) is 180 nM without stacking. Sensitivity can be increased with field-amplified sample stacking by injecting from dilute electrolyte with a detection limit of 19 nM achieved. The high-sensitivity, counter-EOF operation, and short analysis time make this separation well-suited to continuous online monitoring of aerosol composition.

  7. Fully automated microchip system for the detection of quantal exocytosis from single and small ensembles of cells.

    Science.gov (United States)

    Spégel, Christer; Heiskanen, Arto; Pedersen, Simon; Emnéus, Jenny; Ruzgas, Tautgirdas; Taboryski, Rafael

    2008-02-01

    A lab-on-a-chip device that enables positioning of single or small ensembles of cells on an aperture in close proximity to a mercaptopropionic acid (MPA) modified sensing electrode has been developed and characterized. The microchip was used for the detection of Ca(2+)-dependent quantal catecholamine exocytosis from single as well as small assemblies of rat pheochromocytoma (PC12) cells. The frequency of events increased considerably upon depolarization of the PC12 cell membrane using a high extracelluar concentration of potassium. The number of recorded events could be correlated with the number of cells immobilized on the electrode. Quantal characteristics, such as the number of released molecules per recorded event, are equivalent to data obtained using conventional carbon fiber microelectrodes. The detection sensitivity of the device allows for the detection of less than 10 000 dopamine molecules in a quantal release. The distribution of peak rise-time and full width at half maximum was constant during measurement periods of several minutes demonstrating the stability of the MPA modified surface.

  8. Detection of GM soybean by multiplex-touchdown PCR-microchip capillary electrophoresis with response surface methodology optimization.

    Science.gov (United States)

    Li, Yongxin; Su, Ning; Zheng, Bo; Ruan, Jia; Li, Yang; Luo, Chunying; Li, Yuanqian

    2015-02-01

    The combination of the molecular technique, the multivariate strategy and microchip capillary electrophoresis (MCE) was applied to rapid and sensitive analysis of genetically modified (GM) soybean in food samples. A multiplex-touchdown polymerase chain reaction (PCR) system was developed for simultaneously amplifying three target sequences in Roundup Ready soybean (RRS). Response surface methodology was introduced to determine the optimal separation condition in MCE with good resolution and short analytical time. The detection of the PCR products of RRS was completed within 4 min under the optimal conditions. The specificity of the method was evaluated by testing non-GM soybean materials and three GM maize varieties (MON810, Bt176 and Bt11). A sensitivity of 0.1% GM organisms content was obtained, which was remarkably lower than the labeling threshold for transgenic food defined as 0.9% in the European regulation. The relative standard deviation of migration time was in the range of 0.17-0.95%. The proposed method was rapid, sensitive and specific and can be used to identify and detect GM soybean in food samples.

  9. Micellar electrokinetic chromatography on microchips.

    Science.gov (United States)

    Kitagawa, Fumihiko; Otsuka, Koji

    2008-03-01

    This review highlights the methodological and instrumental developments in microchip micellar EKC (MCMEKC) from 1995. The combination of higher separation efficiencies in micellar EKC (MEKC) with high-speed separation in microchip electrophoresis (MCE) should provide high-throughput and high-performance analytical systems. The chip-based separation technique has received considerable attention due to its integration ability without any connector. This advantage allows the development of a multidimensional separation system. Several types of 2-D separation microchips are described in the review. Since complicated channel configurations can easily be fabricated on planar substrates, various sample manipulations can be carried out prior to MCMEKC separations. For example, mixing for on-chip reactions, on-line sample preconcentration, on-chip assay, etc., have been integrated on MEKC microchips. The application of on-line sample preconcentration to MCMEKC can provide not only sensitivity enhancement but also the elucidation of the preconcentration mechanism due to the visualization ability of MCE. The characteristics of these sample manipulations on MEKC microchips are presented in this review. The scope of applications in MCMEKC covers mainly biogenic compounds such as amino acids, peptides, proteins, biogenic amines, DNA, and oestrogens. This review provides a comprehensive table listing the applications in MCMEKC in relation to detection methods.

  10. On-Chip Magnetic Bead Manipulation and Detection Using a Magnetoresistive Sensor-Based Micro-Chip: Design Considerations and Experimental Characterization

    KAUST Repository

    Gooneratne, Chinthaka P.

    2016-08-26

    The remarkable advantages micro-chip platforms offer over cumbersome, time-consuming equipment currently in use for bio-analysis are well documented. In this research, a micro-chip that includes a unique magnetic actuator (MA) for the manipulation of superparamagnetic beads (SPBs), and a magnetoresistive sensor for the detection of SPBs is presented. A design methodology, which takes into account the magnetic volume of SPBs, diffusion and heat transfer phenomena, is presented with the aid of numerical analysis to optimize the parameters of the MA. The MA was employed as a magnetic flux generator and experimental analysis with commercially available COMPEL™ and Dynabeads® demonstrated the ability of the MA to precisely transport a small number of SPBs over long distances and concentrate SPBs to a sensing site for detection. Moreover, the velocities of COMPEL™ and Dynabead® SPBs were correlated to their magnetic volumes and were in good agreement with numerical model predictions. We found that 2.8 μm Dynabeads® travel faster, and can be attracted to a magnetic source from a longer distance, than 6.2 μm COMPEL™ beads at magnetic flux magnitudes of less than 10 mT. The micro-chip system could easily be integrated with electronic circuitry and microfluidic functions, paving the way for an on-chip biomolecule quantification device

  11. On-Chip Magnetic Bead Manipulation and Detection Using a Magnetoresistive Sensor-Based Micro-Chip: Design Considerations and Experimental Characterization

    Directory of Open Access Journals (Sweden)

    Chinthaka P. Gooneratne

    2016-08-01

    Full Text Available The remarkable advantages micro-chip platforms offer over cumbersome, time-consuming equipment currently in use for bio-analysis are well documented. In this research, a micro-chip that includes a unique magnetic actuator (MA for the manipulation of superparamagnetic beads (SPBs, and a magnetoresistive sensor for the detection of SPBs is presented. A design methodology, which takes into account the magnetic volume of SPBs, diffusion and heat transfer phenomena, is presented with the aid of numerical analysis to optimize the parameters of the MA. The MA was employed as a magnetic flux generator and experimental analysis with commercially available COMPEL™ and Dynabeads® demonstrated the ability of the MA to precisely transport a small number of SPBs over long distances and concentrate SPBs to a sensing site for detection. Moreover, the velocities of COMPEL™ and Dynabead® SPBs were correlated to their magnetic volumes and were in good agreement with numerical model predictions. We found that 2.8 μm Dynabeads® travel faster, and can be attracted to a magnetic source from a longer distance, than 6.2 μm COMPEL™ beads at magnetic flux magnitudes of less than 10 mT. The micro-chip system could easily be integrated with electronic circuitry and microfluidic functions, paving the way for an on-chip biomolecule quantification device.

  12. On-Chip Magnetic Bead Manipulation and Detection Using a Magnetoresistive Sensor-Based Micro-Chip: Design Considerations and Experimental Characterization.

    Science.gov (United States)

    Gooneratne, Chinthaka P; Kodzius, Rimantas; Li, Fuquan; Foulds, Ian G; Kosel, Jürgen

    2016-08-26

    The remarkable advantages micro-chip platforms offer over cumbersome, time-consuming equipment currently in use for bio-analysis are well documented. In this research, a micro-chip that includes a unique magnetic actuator (MA) for the manipulation of superparamagnetic beads (SPBs), and a magnetoresistive sensor for the detection of SPBs is presented. A design methodology, which takes into account the magnetic volume of SPBs, diffusion and heat transfer phenomena, is presented with the aid of numerical analysis to optimize the parameters of the MA. The MA was employed as a magnetic flux generator and experimental analysis with commercially available COMPEL™ and Dynabeads(®) demonstrated the ability of the MA to precisely transport a small number of SPBs over long distances and concentrate SPBs to a sensing site for detection. Moreover, the velocities of COMPEL™ and Dynabead(®) SPBs were correlated to their magnetic volumes and were in good agreement with numerical model predictions. We found that 2.8 μm Dynabeads(®) travel faster, and can be attracted to a magnetic source from a longer distance, than 6.2 μm COMPEL™ beads at magnetic flux magnitudes of less than 10 mT. The micro-chip system could easily be integrated with electronic circuitry and microfluidic functions, paving the way for an on-chip biomolecule quantification device.

  13. Microchip CE analysis of amino acids on a titanium dioxide nanoparticles-coated PDMS microfluidic device with in-channel indirect amperometric detection.

    Science.gov (United States)

    Qiu, Jian-Ding; Wang, Li; Liang, Ru-Ping; Wang, Jing-Wu

    2009-10-01

    In this paper, titanium dioxide nanoparticles (TiO(2) NPs) were employed to construct a functional film on PDMS microfluidic channel surface, which was formed by sequentially immobilizing poly(diallyldimethylammonium chloride) and TiO(2) NPs on PDMS surface by layer-by-layer assembly technique. The modified PDMS microchip exhibited a decreased and stable EOF, which was favorable for the separation of biomolecules with similar migration times. Arginine, phenylalanine, serine and threonine were used as model analytes to evaluate the performance of the modified microchip. The four amino acids were efficiently separated within 100 s in a 3.7 cm long separation channel and successfully detected on the carbon fiber electrode in conjunction with in-channel indirect amperometry. Resolutions and theoretical plate numbers of the analytes were considerably enhanced in the presence of TiO(2) NPs. The modified microchip demonstrated excellent stability and reproducibility with improved RSDs of migration times and peak currents for run-to-run, day-to-day and chip-to-chip analyses, respectively. Variables influencing the separation efficiency and amperometric response, including injection and separation voltage, the working electrode position and buffer concentration, were optimized in detail.

  14. Electronic DNA detection and diagnostics

    NARCIS (Netherlands)

    De, Arpita

    2013-01-01

    The Nanopill project is an ambitious undertaking with the objective to develop an early-warning cancer diagnostic pill that is ingested by the patient. The Nanopill collects intestinal fluid as the pill travels down the intestinal tract, and tests for the presence of a free floating hyper-methylated

  15. Nanostructure embedded microchips for detection, isolation, and characterization of circulating tumor cells.

    Science.gov (United States)

    Lin, Millicent; Chen, Jie-Fu; Lu, Yi-Tsung; Zhang, Yang; Song, Jinzhao; Hou, Shuang; Ke, Zunfu; Tseng, Hsian-Rong

    2014-10-21

    Circulating tumor cells (CTCs) are cancer cells that break away from either a primary tumor or a metastatic site and circulate in the peripheral blood as the cellular origin of metastasis. With their role as a "tumor liquid biopsy", CTCs provide convenient access to all disease sites, including that of the primary tumor and the site of fatal metastases. It is conceivable that detecting and analyzing CTCs will provide insightful information in assessing the disease status without the flaws and limitations encountered in performing conventional tumor biopsies. However, identifying CTCs in patient blood samples is technically challenging due to the extremely low abundance of CTCs among a large number of hematologic cells. To address this unmet need, there have been significant research endeavors, especially in the fields of chemistry, materials science, and bioengineering, devoted to developing CTC detection, isolation, and characterization technologies. Inspired by the nanoscale interactions observed in the tissue microenvironment, our research team at UCLA pioneered a unique concept of "NanoVelcro" cell-affinity substrates, in which CTC capture agent-coated nanostructured substrates were utilized to immobilize CTCs with high efficiency. The working mechanism of NanoVelcro cell-affinity substrates mimics that of Velcro: when the two fabric strips of a Velcro fastener are pressed together, tangling between the hairy surfaces on two strips leads to strong binding. Through continuous evolution, three generations (gens) of NanoVelcro CTC chips have been established to achieve different clinical utilities. The first-gen NanoVelcro chip, composed of a silicon nanowire substrate (SiNS) and an overlaid microfluidic chaotic mixer, was created for CTC enumeration. Side-by-side analytical validation studies using clinical blood samples suggested that the sensitivity of first-gen NanoVelcro chip outperforms that of FDA-approved CellSearch. In conjunction with the use of the

  16. Indirect detection of superoxide in RAW 264.7 macrophage cells using microchip electrophoresis coupled to laser-induced fluorescence.

    Science.gov (United States)

    de Campos, Richard P S; Siegel, Joseph M; Fresta, Claudia G; Caruso, Giuseppe; da Silva, José A F; Lunte, Susan M

    2015-09-01

    Superoxide, a naturally produced reactive oxygen species (ROS) in the human body, is involved in many pathological and physiological signaling processes. However, if superoxide formation is left unregulated, overproduction can lead to oxidative damage to important biomolecules, such as DNA, lipids, and proteins. Superoxide can also lead to the formation of peroxynitrite, an extremely hazardous substance, through its reaction with endogenously produced nitric oxide. Despite its importance, quantitative information regarding superoxide production is difficult to obtain due to its high reactivity and low concentrations in vivo. MitoHE, a fluorescent probe that specifically reacts with superoxide, was used in conjunction with microchip electrophoresis (ME) and laser-induced fluorescence (LIF) detection to investigate changes in superoxide production by RAW 264.7 macrophage cells following stimulation with phorbol 12-myristate 13-acetate (PMA). Stimulation was performed in the presence and absence of the superoxide dismutase (SOD) inhibitors, diethyldithiocarbamate (DDC) and 2-metoxyestradiol (2-ME). The addition of these inhibitors resulted in an increase in the amount of superoxide specific product (2-OH-MitoE(+)) from 0.08 ± 0.01 fmol (0.17 ± 0.03 mM) in native cells to 1.26 ± 0.06 fmol (2.5 ± 0.1 mM) after PMA treatment. This corresponds to an approximately 15-fold increase in intracellular concentration per cell. Furthermore, the addition of 3-morpholino-sydnonimine (SIN-1) to the cells during incubation resulted in the production of 0.061 ± 0.006 fmol (0.12 ± 0.01 mM) of 2-OH-MitoE(+) per cell on average. These results demonstrate that indirect superoxide detection coupled with the use of SOD inhibitors and a separation method is a viable method to discriminate the 2-OH-MitoE(+) signal from possible interferences.

  17. A novel CE microchip with micro pillars column & double-L injection design for Capacitance Coupled Contactless Conductivity detection technology

    Science.gov (United States)

    Wang, Yineng; Messina, Walter; Cao, Xi; Hogan, Anna; van Zalen, Ed; Moore, Eric

    2016-10-01

    This novel capillary electrophoresis microchip, or also known as μTAS (micro total analysis system) was designed to separate complex aqueous based compounds, similar to commercial CE & microchip (capillary electrophoresis) systems, but more compact. This system can be potentially used for mobile/portable chemical analysis equipment. Un-doped silicon wafer & ultra-thin borofloat glass (Pyrex) wafers have been used to fabricate the device. Double-L injection feature, micro pillars column, bypass separation channel & hybrid- referenced C4D electrodes were designed to achieve a high SNR (signal to noise ratio), easy- separation, for a durable and reusable μTAS for CE use.

  18. Fast and simultaneous detection of heavy metals using a simple and reliable microchip-electrochemistry route: An alternative approach to food analysis.

    Science.gov (United States)

    Chailapakul, Orawon; Korsrisakul, Sarawadee; Siangproh, Weena; Grudpan, Kate

    2008-01-15

    This paper reports, for the first, the fast and simultaneous detection of prominent heavy metals, including: lead, cadmium and copper using microchip CE with electrochemical detection. The direct amperometric detection mode for microchip CE was successfully applied to these heavy metal ions. The influences of separation voltage, detection potential, as well as the concentration and pH value of the running buffer on the response of the detector were carefully assayed and optimized. The results clearly show that reliable analysis for lead, cadmium, and copper by the degree of electrophoretic separation occurs in less than 3min using a MES buffer (pH 7.0, 25mM) and l-histidine, with 1.2kV separation voltage and -0.8V detection potential. The detection limits for Pb(2+), Cd(2+), and Cu(2+) were 1.74, 0.73 and 0.13microM (S/N=3). The %R.S.D. of each peak current was foods.

  19. Doping of a dielectric layer as a new alternative for increasing sensitivity of the contactless conductivity detection in microchips.

    Science.gov (United States)

    Lima, Renato Sousa; Segato, Thiago Pinotti; Gobbi, Angelo Luiz; Coltro, Wendell Karlos Tomazelli; Carrilho, Emanuel

    2011-12-21

    This communication describes a new procedure to increase the sensitivity of C(4)D in PDMS/glass microchips. The method consists in doping the insulating layer (PDMS) over the electrodes with nanoparticles of TiO(2), increasing thus its dielectric constant. The experimental protocol is simple, inexpensive, and fast.

  20. A low-density DNA microchip for the detection of (anti-)estrogenic compounds and their relative potencies

    NARCIS (Netherlands)

    Wang, S.; Rijk, J.C.W.; Pen, M.J.; Aarts, J.M.; Peijnenburg, A.A.C.M.; Rietjens, I.; Bovee, T.F.H.

    2013-01-01

    In the current study, a set of 12 reference compounds was tested in a low-density DNA microchip that contains probes for 11 different estrogen-responsive marker genes. Our results show that the seven most informative marker genes on the chip resulted in fingerprints that correctly predicted the (ant

  1. Monitoring of nitrite, nitrate, chloride and sulfate in environmental samples using electrophoresis microchips coupled with contactless conductivity detection.

    Science.gov (United States)

    Freitas, Camilla Benevides; Moreira, Roger Cardoso; de Oliveira Tavares, Maria Gizelda; Coltro, Wendell K T

    2016-01-15

    This report describes the development of an analytical methodology on microchip electrophoresis (ME) devices coupled with capacitively coupled contactless conductivity detection (C(4)D) to monitor inorganic anions in environmental samples. The buffer composition as well as detection operating parameters were optimized to achieve the best separation selectivity and detector sensitivity, respectively. Electrophoretic separations of Cl(-), NO3(-), SO4(2-) and NO2(-) were successfully performed within 60s using a running buffer composed of 30mmol L(-1) latic acid and 15mmol L(-1)l-histidine (His). The best detectability levels were found applying a sinusoidal wave with 1100-kHz-frequency and 60-Vpp amplitude. Quantitative analyzes of inorganic anions were carried out in the presence of Cr2O7(2-) ion as internal standard (IS), which ensured great repeatability in terms of migration times (<1%) and peak areas (6.2-7.6%) for thirty consecutive injections. The analytical performance revealed a linear behavior for concentration ranges between 0-120μmol L(-1) (Cl(-), NO2(-) and NO3(-)) and 0-60μmol L(-1) (SO4(2-)) and limits of detection (LODs) varying from 2.0 to 4.9μmol L(-1). The concentration levels of anionic species were determined in aquarium, river and biofertilizer samples with recovery values between 91% and 105%. The nitrification steps associated with conversion of ammonium to nitrite followed by the conversion of nitrite to nitrate were successfully monitored in a simulated environment without fishes during a period of twelve weeks. Lastly, the monitoring of anionic species was carried out during eight weeks in an aquarium environment containing ten fishes from Danio rerio (Ciprynidae). The recorded data revealed the absence of nitrite and a gradual increase on the ammonium and nitrate concentration levels during eight weeks, thus suggesting the direct conversion of ammonium to nitrate. Based on the data herein reported, the proposed analytical methodology

  2. Ultratrace DNA Detection Based on the Condensing-Enrichment Effect of Superwettable Microchips.

    Science.gov (United States)

    Xu, Li-Ping; Chen, Yanxia; Yang, Gao; Shi, Wanxin; Dai, Bing; Li, Guannan; Cao, Yanhua; Wen, Yongqiang; Zhang, Xueji; Wang, Shutao

    2015-11-18

    A sensitive nucleic acid detection platform based on superhydrophilic microwells spotted on a superhydrophobic substrate is fabricated. Due to the wettability differences, ultratrace DNA molecules are enriched and the fluorescent signals are amplified to allow more sensitive detection. The biosensing interface based on superwettable materials provides a simple and cost-effective way for ultratrace DNA sensing.

  3. Real-Time PCR using a PCR Microchip with Integrated Thermal System and Polymer Waveguides for the Detection of Campylobacter jejuni

    DEFF Research Database (Denmark)

    Wang, Zhenyu; Sekulovic, Andrea; Kutter, Jörg Peter

    2006-01-01

    A novel real-time PCR microchip platform with integrated thermal system and polymer waveguides has been developed. By using the integrated optical system of the real-time PCR chip, cadF – a virulence gene of Campylobacter jejuni, could specifically be detected. Two different DNA binding dyes, SYTOX...... Orange and TO-PRO-3, were added to the PCR mixture to realize the real-time PCR. The presented approach shows reliable real-time quantitative information of the PCR amplification of the targeted gene....

  4. Microchip-based ELISA strategy for the detection of low-level disease biomarker in serum

    Energy Technology Data Exchange (ETDEWEB)

    Liu Yun; Wang Huixiang; Huang Jingyu; Yang Jie; Liu Baohong [Department of Chemistry, Institute of Biomedical Sciences, Fudan University, Shanghai 200433 (China); Yang Pengyuan, E-mail: pyyang@fudan.edu.cn [Department of Chemistry, Institute of Biomedical Sciences, Fudan University, Shanghai 200433 (China)

    2009-09-14

    A simple and sensitive method has been proposed to determine a trace level of {alpha}-fetoprotein (AFP), hepatocellular carcinoma biomarker, using poly(methyl methacrylate) (PMMA) microfluidic chips coupled with electrochemical detection system. The PMMA microchannels have been modified with poly(ethyleneimine) (PEI) containing abundant NH{sub 2} groups to covalently immobilize AFP monoclonal antibody. Afterward, the antigen AFP and horseradish peroxidase (HRP)-conjugated AFP antibody can sequentially bind through antigen-antibody specific interaction. Atomic force microscopy (AFM) and confocal fluorescence microscope (CFFM) were utilized to characterize the surface topography and protein immobilization after modification. Coupled with three-electrode electrochemical detection system, the immunochip can perform the detection limit of AFP down to 1 pg mL{sup -1}, and achieve a detectable linear concentration range of 1-500 pg mL{sup -1} by differential pulse voltammetry (DPV). The on-chip immunoassay platform can not only provide rapid and sensitive detection for target proteins but also be resistant to non-specific adsorption of proteins, which contributes to the detection of low-level protein in real sample. Finally, AFP existing in healthy human serum was detected to demonstrate the utility of the immunochip. The result shows that the proposed approach is feasible and has the potential application in clinical analysis and diagnosis.

  5. Separation of natural antioxidants using PDMS electrophoresis microchips coupled with amperometric detection and reverse polarity.

    Science.gov (United States)

    Lucca, Bruno Gabriel; Lunte, Susan Marie; Tomazelli Coltro, Wendell Karlos; Ferreira, Valdir Souza

    2014-12-01

    This report describes the use of PDMS ME coupled with amperometric detection for rapid separation of ascorbic, gallic , ferulic, p-coumaric acids using reverse polarity. ME devices were fabricated in PDMS by soft lithography and detection was accomplished using an integrated carbon fiber working electrode aligned in the end-channel configuration. Separation and detection parameters were investigated and the best conditions were obtained using a run buffer consisting of 5 mM phosphate buffer (pH 6.9) and a detection voltage of 1.0 V versus Ag/AgCl reference electrode. All compounds were separated within 70 s using gated injection mode with baseline resolution and separation efficiencies between 1200 and 9000 plates. Calibration curves exhibited good linearity and the LODs achieved ranged from 1.7 to 9.7 μM. The precision for migration time and peak height provided maximum values of 4% for the intrachip studies. Lastly, the analytical method was successfully applied for the analysis of ascorbic and gallic acids in commercial beverage samples. The results achieved using ME coupled with amperometric detection were in good agreement with the values provided by the supplier. Based on the data reported here, the proposed method shows suitability to be applied for the routine analysis of beverage samples.

  6. Recent advances in chemiluminescence detection coupled with capillary electrophoresis and microchip capillary electrophoresis.

    Science.gov (United States)

    Liu, Yuxuan; Huang, Xiangyi; Ren, Jicun

    2016-01-01

    CE is an ideal analytical method for extremely volume-limited biological microenvironments. However, the small injection volume makes it a challenge to achieve highly sensitive detection. Chemiluminescence (CL) detection is characterized by providing low background with excellent sensitivity because of requiring no light source. The coupling of CL with CE and MCE has become a powerful analytical method. So far, this method has been widely applied to chemical analysis, bioassay, drug analysis, and environment analysis. In this review, we first introduce some developments for CE-CL and MCE-CL systems, and then put the emphasis on the applications in the last 10 years. Finally, we discuss the future prospects.

  7. Measurement of Nitrogen Mustard Degredation Products by Poly(dimethylsiloxane) Microchip Electrophoresis with Contactless Conductivity Detection

    Science.gov (United States)

    The potential risk of human exposure from an accidental or intentional release of CWAs into a civilian population continues to drive the need for screening and monitoring techniques for these compounds. In particular, rapid and reliable methods for detecting CWAs such as the nitr...

  8. [Microchips based on three dimensional gel cells: history and perspective].

    Science.gov (United States)

    Kolchinskiĭ, A M; Griadunov, D A; Lysov, Iu P; Mikhaĭlovich, V M; Nasedkina, T V; Turygin, A Iu; Rubina, A Iu; Barskiĭ, V E; Zasedatelev, A S

    2004-01-01

    The review describes the history of creation and development of the microchip technology and its role in the human genome project in Russia. The emphasis is placed on the three-dimensional gel-based microchips developed at the Center of Biological Microchips headed by A.D. Mirzabekov since 1988. The gel-based chips of the last generation, IMAGE chips (Immobilized Micro Array of Gel Elements), have a number of advantages over the previous versions. The microchips are manufactured by photo-initiated copolymerization of gel components and immobilized molecules (DNA, proteins, and ligands). This ensures an even distribution of the immobilized probe throughout the microchip gel element with a high yield (about 50% for oligonucleotides). The use of methacrylamide as a main component of the polymerization mixture resulted in a substantial increase of gel porosity without affecting its mechanical strength and stability, which allowed one to work with the DNA fragments of up to 500 nt in length, as well as with rather large protein molecules. At present, the gel-based microchips are widely applied to address different problems. The generic microchips containing a complete set of possible hexanucleotides are used to reveal the DNA motifs binding with different proteins and to study the DNA-protein interactions. The oligonucleotide microchips are a cheap and reliable tool of diagnostics designed for mass application. Biochips have been developed for identification of the tuberculosis pathogen and its antibiotic-resistant forms; for diagnostics of orthopoxviruses, including the smallpox virus; for diagnostics of the anthrax pathogen; and for identification of chromosomal rearrangements in leukemia patients. The protein microchips can be adapted for further use in proteomics. Bacterial and yeast cells were also immobilized in the gel, maintaining their viability, which open a wide potential for creation biosensors on the basis of microchips.

  9. The Next Generation MOD: A Microchip Amino Acid Analyzer for Detecting Extraterrestrial Life

    Science.gov (United States)

    Mathies, R. A.; Hutt, L. D.; Bada, J. L.; Glavin, D.; Grunthaner, F. J.; Grunthaner, P. J.

    2000-01-01

    The MOD (Mars Organic Detector) instrument which has selected for the definition phase of the BEDS package on the 2005 Mars Explorer Program spacecraft is designed to simply detect the presence of amino acids in Martian surface samples at a sensitivity of a few parts per billion (ppb). An additional important aspect of amino acid analyses of Martian samples is identifying and quantifying which compounds are present, and also distinguishing those produced abiotically from those synthesized by either extinct or extant life. Amino acid homochirality provides an unambiguous way of distinguishing between abiotic vs. biotic origins. Proteins made up of mixed D- and L-amino acids would not likely have been efficient catalysts in early organisms because they could not fold into bioactive configurations such as the a-helix. However, enzymes made up of all D-amino acids function just as well as those made up of only L-amino acids, but the two enzymes use the opposite stereoisomeric substrates. There are no biochemical reasons why L-amino acids would be favored over Damino acids. On Earth, the use of only L-amino acids in proteins by life is probably simply a matter of chance. We assume that if proteins and enzymes were a component of extinct or extant life on Mars, then amino acid homochirality would have been a requirement. However, the possibility that Martian life was (or is) based on D-amino acids would be equal to that based on L-amino acids. The detection of a nonracemic mixture of amino acids in a Martian sample would be strong evidence for the presence of an extinct or extant biota on Mars. The finding of an excess of D-amino acids would provide irrefutable evidence of unique Martian life that could not have been derived from seeding the planet with terrestrial life (or the seeding of the Earth with Martian life). In contrast, the presence of racemic amino acids, along with non-protein amino acids such as alpha-aminoisobutyric acid and isovaline, would be indicative

  10. The Next Generation MOD: A Microchip Amino Acid Analyzer for Detecting Extraterrestrial Life

    Science.gov (United States)

    Mathies, R. A.; Hutt, L. D.; Bada, J. L.; Glavin, D.; Grunthaner, F. J.; Grunthaner, P. J.

    2000-01-01

    The MOD (Mars Organic Detector) instrument which has selected for the definition phase of the BEDS package on the 2005 Mars Explorer Program spacecraft is designed to simply detect the presence of amino acids in Martian surface samples at a sensitivity of a few parts per billion (ppb). An additional important aspect of amino acid analyses of Martian samples is identifying and quantifying which compounds are present, and also distinguishing those produced abiotically from those synthesized by either extinct or extant life. Amino acid homochirality provides an unambiguous way of distinguishing between abiotic vs. biotic origins. Proteins made up of mixed D- and L-amino acids would not likely have been efficient catalysts in early organisms because they could not fold into bioactive configurations such as the a-helix. However, enzymes made up of all D-amino acids function just as well as those made up of only L-amino acids, but the two enzymes use the opposite stereoisomeric substrates. There are no biochemical reasons why L-amino acids would be favored over Damino acids. On Earth, the use of only L-amino acids in proteins by life is probably simply a matter of chance. We assume that if proteins and enzymes were a component of extinct or extant life on Mars, then amino acid homochirality would have been a requirement. However, the possibility that Martian life was (or is) based on D-amino acids would be equal to that based on L-amino acids. The detection of a nonracemic mixture of amino acids in a Martian sample would be strong evidence for the presence of an extinct or extant biota on Mars. The finding of an excess of D-amino acids would provide irrefutable evidence of unique Martian life that could not have been derived from seeding the planet with terrestrial life (or the seeding of the Earth with Martian life). In contrast, the presence of racemic amino acids, along with non-protein amino acids such as alpha-aminoisobutyric acid and isovaline, would be indicative

  11. Microchips on glass

    NARCIS (Netherlands)

    Keulemans, M.

    2007-01-01

    Microchips on glass. What about a mobile phone that uses a single microchip to receive all the available frequency bands, plus extras such as television, gps, and Internet access? Or, in due time, see-though implants that will monitor your state of health, and equipment that will let you see through

  12. Development of an SDS-gel electrophoresis method on SU-8 microchips for protein separation with LIF detection: Application to the analysis of whey proteins.

    Science.gov (United States)

    Del Mar Barrios-Romero, Maria; Crevillén, Agustín G; Diez-Masa, José Carlos

    2013-08-01

    This work describes the development of an SDS-gel electrophoresis method for the analysis of major whey proteins (α-lactalbumin, β-lactoglobulin, and BSA) carried out in SU-8 microchips. The method uses a low-viscosity solution of dextran as a sieving polymer. A commercial coating agent (EOTrol LN) was added to the separation buffer to control the EOF of the chips. The potential of this coating agent to prevent protein adsorption on the walls of the SU-8 channels was also evaluated. Additionally, the fluorescence background of the SU-8 material was studied to improve the sensitivity of the method. By selecting an excitation wavelength of 532 nm at which the background fluorescence remains low and by replacing the mercury arc lamp by a laser in the detection system, an LOD in the nanomolar range was achieved for proteins derivatized with the fluorogenic reagent Chromeo P540. Finally, the method was applied to the analysis of milk samples, demonstrating the potential of SU-8 microchips for the analysis of proteins in complex food samples.

  13. Microchip in situ electrosynthesis of silver metallic oxide clusters for ultra-FAST detection of galactose in galactosemic newborns' urine samples.

    Science.gov (United States)

    García-Carmona, Laura; Rojas, Daniel; González, María Cristina; Escarpa, Alberto

    2016-10-17

    This work describes for the first time the coupling of microfluidic chips (MC) to electrosynthetized silver metallic oxide clusters (AgMOCs). As an early demonstration of this novel approach, the ultrafast detection of galactose in galactosemic newborns' urine samples is proposed. AgMOCs were in situ electrosynthetized on integrated microchip platinum electrodes using a double pulse technique and characterized in full using scanning electronic microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), X-ray photoelectron spectroscopy (XPS) and electrochemical techniques revealing the presence of silver oxides and electrocatalysis towards galactose as a galactosemia biomarker. Galactose detection in galactosemic newborns' urine samples proceeded in less than 30 s, differentiating between ill and healthy urine samples and requiring negligible urine sample consumption. The significance of the newborns' urine samples confirmed the analytical potency of the MC-AgMOCs approach for future implementation of screening for rare disease diagnosis such as galactosemia.

  14. A low-density DNA microchip for the detection of (anti-)estrogenic compounds and their relative potencies.

    Science.gov (United States)

    Wang, Si; Rijk, Jeroen C W; Pen, Marieke J; Aarts, Jac M M J G; Peijnenburg, Ad A C M; Rietjens, Ivonne M C M; Bovee, Toine F H

    2013-04-01

    In the current study, a set of 12 reference compounds was tested in a low-density DNA microchip that contains probes for 11 different estrogen-responsive marker genes. Our results show that the seven most informative marker genes on the chip resulted in fingerprints that correctly predicted the (anti-)estrogenic activity of the model compounds except that of the negative control testosterone. Two marker genes, myeloid leukemia factor-1 interacting protein and ubiquitin-conjugating enzyme E2C, were even capable of correctly predicting the estrogenic potency of all five estrogen receptor (ER) agonists tested and correlated well with the potencies as determined in the MCF-7/BOS proliferation assay and the in vivo uterotrophic assay. In addition, it was demonstrated that the estrogenic responses of testosterone, both in the array tube assay and in the proliferation assay, were partially due to the conversion of testosterone into 17β-estradiol by aromatase but also due to formation of other estrogenic metabolites, the presence and estrogenic potency of which were confirmed by gas chromatography-tandem mass spectrometry analysis and a yeast-based reporter gene assay, respectively. It is concluded that low-density DNA microchip-based fingerprinting in MCF-7/BOS cells for estrogenicity marker genes provides a faster in vitro alternative to the current MCF-7/BOS cell proliferation assay (E-screen).

  15. Photonic Crystal Microchip Laser

    Science.gov (United States)

    Gailevicius, Darius; Koliadenko, Volodymyr; Purlys, Vytautas; Peckus, Martynas; Taranenko, Victor; Staliunas, Kestutis

    2016-09-01

    The microchip lasers, being very compact and efficient sources of coherent light, suffer from one serious drawback: low spatial quality of the beam strongly reducing the brightness of emitted radiation. Attempts to improve the beam quality, such as pump-beam guiding, external feedback, either strongly reduce the emission power, or drastically increase the size and complexity of the lasers. Here it is proposed that specially designed photonic crystal in the cavity of a microchip laser, can significantly improve the beam quality. Experiments show that a microchip laser, due to spatial filtering functionality of intracavity photonic crystal, improves the beam quality factor M2 reducing it by a factor of 2, and increase the brightness of radiation by a factor of 3. This comprises a new kind of laser, the “photonic crystal microchip laser”, a very compact and efficient light source emitting high spatial quality high brightness radiation.

  16. Controlled-release microchips.

    Science.gov (United States)

    Sharma, Sadhana; Nijdam, A Jasper; Sinha, Piyush M; Walczak, Robbie J; Liu, Xuewu; Cheng, Mark M-C; Ferrari, Mauro

    2006-05-01

    Efficient drug delivery remains an important challenge in medicine: continuous release of therapeutic agents over extended time periods in accordance with a predetermined temporal profile; local delivery at a constant rate to the tumour microenvironment to overcome much of the systemic toxicity and to improve antitumour efficacy; improved ease of administration, and increasing patient compliance required are some of the unmet needs of the present drug delivery technology. Microfabrication technology has enabled the development of novel controlled-release microchips with capabilities not present in the current treatment modalities. In this review, the current status and future prospects of different types of controlled-release microchips are summarised and analysed with reference to microneedle-based microchips, as well as providing an in-depth focus on microreservoir-based and nanoporous microchips.

  17. Recent innovations in protein separation on microchips by electrophoretic methods.

    Science.gov (United States)

    Peng, Youyuan; Pallandre, Antoine; Tran, N Thuy; Taverna, Myriam

    2008-01-01

    Microchips for analytical purposes have attracted great attention over the last 20 years. In the present review, we focus on the most recent development of microchips for electrophoretic separation of proteins. This review starts with a short recalling about the microchips covering the basic microchip layout for CE and the commercial chips and microchip platforms. A short paragraph is dedicated to the surface treatment of microchips, which is of paramount importance in protein analysis. One section is dedicated to on-line sample pretreatment in microchips and summarizes different strategies to pre-concentrate or to purify proteins from complex matrixes. Most of the common modes used for CE of proteins have already been adapted to the chip format, while multidimensional approaches are still in progress. The different routes to achieve detection in microchip are also presented with a special attention to derivatization or labeling of proteins. Finally, several recent applications are mentioned. They highlight the great potential of electrophoretic separations of proteins in numerous fields such as biological, pharmaceutical or agricultural and food analysis. A bibliography with 151 references is provided covering papers published from 2000 to the early 2007.

  18. Diagnostic Procedures to Detect Chlamydia trachomatis Infections.

    Science.gov (United States)

    Meyer, Thomas

    2016-08-05

    The intracellular life style of chlamydia and the ability to cause persistent infections with low-grade replication requires tests with high analytical sensitivity to directly detect C. trachomatis (CT) in medical samples. Nucleic acid amplification tests (NAATs) are the most sensitive assays with a specificity similar to cell culture and are considered the method of choice for CT detection. In addition, NAATs can be performed on various clinical specimens that do not depend on specific transport and storage conditions, since NAATs do not require infectious bacteria. In the case of lower genital tract infections, first void urine and vaginal swabs are the recommended specimens for testing males and females, respectively. Infections of anorectal, oropharyngeal and ocular epithelia should also be tested by NAAT analysis of corresponding mucosal swabs. In particular, anorectal infections of men who have sex with men (MSM) should include evaluation of lymphogranuloma venereum (LGV) by identification of genotypes L1, L2 or L3. Detection of CT antigens by enzyme immunoassay (EIAs) or rapid diagnostic tests (RDTs) are unsuitable due to insufficient sensitivity and specificity. Recent PCR-based RDTs, however, are non-inferior to standard NAATs, and might be used at the point-of-care. Serology finds application in the diagnostic work-up of suspected chronic CT infection but is inappropriate to diagnose acute infections.

  19. Evaluation of in-channel amperometric detection using a dual-channel microchip electrophoresis device and a two-electrode potentiostat for reverse polarity separations.

    Science.gov (United States)

    Meneses, Diogenes; Gunasekara, Dulan B; Pichetsurnthorn, Pann; da Silva, José A F; de Abreu, Fabiane C; Lunte, Susan M

    2015-02-01

    In-channel amperometric detection combined with dual-channel microchip electrophoresis is evaluated using a two-electrode isolated potentiostat for reverse polarity separations. The device consists of two separate channels with the working and reference electrodes placed at identical positions relative to the end of the channel, enabling noise subtraction. In previous reports of this configuration, normal polarity and a three-electrode detection system were used. In the two-electrode detection system described here, the electrode in the reference channel acts as both the counter and reference. The effect of electrode placement in the channels on noise and detector response was investigated using nitrite, tyrosine, and hydrogen peroxide as model compounds. The effects of electrode material and size and type of reference electrode on noise and the potential shift of hydrodynamic voltammograms for the model compounds were determined. In addition, the performance of two- and three-electrode configurations using Pt and Ag/AgCl reference electrodes was compared. Although the signal was attenuated with the Pt reference, the noise was also significantly reduced. It was found that lower LOD were obtained for all three compounds with the dual-channel configuration compared to single-channel, in-channel detection. The dual-channel method was then used for the detection of nitrite in a dermal microdialysis sample obtained from a sheep following nitroglycerin administration.

  20. Enhancing the performance of a point-of-care CD4+ T-cell counting microchip through monocyte depletion for HIV/AIDS diagnostics.

    Science.gov (United States)

    Cheng, Xuanhong; Gupta, Amit; Chen, Chihchen; Tompkins, Ronald G; Rodriguez, William; Toner, Mehmet

    2009-05-21

    CD4+ T cell counts are important tests used to stage HIV-positive patients, enabling clinicians to make informed antiretroviral treatment decisions and to monitor the therapeutic outcomes. However, state-of-the-art CD4 counting methods based on flow cytometry are not applicable in resource-limited settings, due to their high cost and technical requirements. In previous work, we reported the development of a cell isolation microchip that can be used at the point of care for CD4 counts. In that microfluidic chip, CD4+ T cells were separated from 10 microL of whole blood, and enumerated via either light microscopy or impedance sensing. The microchip counts matched flow cytometry results in the intermediate CD4 count range, between 200-800 cells/microL, but displayed a positive bias at absolute CD4 counts below 200 cells/microL, due largely to monocyte contamination. To enhance the performance in the low CD4 count range, we report here an improved design of a two-stage microfluidic device to deplete monocytes from whole blood, followed by CD4+ T cell capture. Using the double-stage device combined with a high viscosity rinsing solution, we obtained microchip CD4 counts comparable to flow cytometry results in the full clinically relevant range. In addition to CD4 counting, the strategy of contaminant depletion prior to target cell isolation can be easily adapted to immunoaffinity capture of other cell types that lack a unique surface marker from a complex biological fluid.

  1. Electrochemistry-based real-time PCR on a microchip.

    Science.gov (United States)

    Yeung, Stephen S W; Lee, Thomas M H; Hsing, I-Ming

    2008-01-15

    The development of handheld instruments for point-of-care DNA analysis can potentially contribute to the medical diagnostics and environmental monitoring for decentralized applications. In this work, we demonstrate the implementation of a recently developed electrochemical real-time polymerase chain reaction (ERT-PCR) technique on a silicon-glass microchip for simultaneous DNA amplification and detection. This on-chip ERT-PCR process requires the extension of an oligonucleotide in both solution and at solid phases and intermittent electrochemical signal measurement in the presence of all the PCR reagents. Several important parameters, related to the surface passivation and electrochemical scanning of working electrodes, were investigated. It was found that the ERT-PCR's onset thermal cycle ( approximately 3-5), where the analytical signal begins to be distinguishable from the background, is much lower than that of the fluorescence-based counterparts for high template DNA situations (3 x 10(6) copies/microL). By carefully controlling the concentrations of the immobilized probe and the enzyme polymerase, improvements have been made in obtaining a meaningful electrochemical signal using a lower initial template concentration. This ERT-PCR technique on a microchip platform holds significant promise for rapid DNA detection for point-of-care testing applications.

  2. Photometric flow injection determination of phosphate on a PDMS microchip using an optical detection system assembled with an organic light emitting diode and an organic photodiode.

    Science.gov (United States)

    Liu, Rong; Ishimatsu, Ryoichi; Yahiro, Masayuki; Adachi, Chihaya; Nakano, Koji; Imato, Toshihiko

    2015-01-01

    A compact photometric detector was constructed from an organic light emitting diode (OLED) based on a europium complex, europium(diben-zoylmethanato)3(bathophenanthroline) (Eu(DBM)3bath), as the light source and an organic photodiode (OPD) fabricated from a hetero-junction of two layers of copper phthalocyanine (CuPc)/fullerene (C60) as the photo-detector on a microchip prepared from poly(dimethylsiloxan) (PDMS) and was applied to the determination of phosphate. The OLED and the OPD were fabricated by a vapor deposition method on an indium tin oxide (ITO) coated glass substrate with the following layered structure; Glass (0.7 mm)/ITO (110 nm)/4,4'-bis[N-(1-naphthyl)-N-phenyl amino]-biphenyl (α-NPD) (30 nm)/4,4'-di(N-carbazolyl)biphenyl (CBP): Eu(3+) (8 wt%, 30 nm)/bathocuproine (BCP) (30 nm)/aluminum tris(8-hydroxyquinoline) (Alq3) (25 nm)/magnesium and silver (MgAg) (100 nm)/Ag (10nm) and Glass (0.7 mm)/ITO (110 nm)/CuPc (35 nm)/C60 (50 nm)/BCP (10 nm)/Ag (50 nm), respectively. The OLED based on the europium complex emitted a sharp light at the wavelength of 612 nm with a full width at half maximum (FWHM) of 8 nm. The performance of the photometric detector assembled was evaluated based on measurements of the absorbance of different concentrations of malachite green (MG) solutions for a batch system with 1cm long path length. The molar absorptive coefficient of the MG solution, calculated from the photocurrent of the OPD, was in good agreement with the value reported in the literature. A microchip with two inlets and one outlet U-shaped channel was prepared by a conventional photolithograph method. The OLED and the OPD were configured so as to face each other through the PDMS microchip in parallel in order to align the light axis of the OLED and the OPD with the flow cell (optical path length of 5mm), which was located at the end of outlet. For the determination of phosphate, an ion-association reaction between MG and a molybdenum-phosphate complex was utilized

  3. Coherence in Microchip Traps

    CERN Document Server

    Treutlein, P; Steinmetz, T; Hänsch, T W; Reichel, J; Treutlein, Philipp; Hommelhoff, Peter; Steinmetz, Tilo; H\\"ansch, Theodor W.; Reichel, Jakob

    2003-01-01

    We report the coherent manipulation of internal states of neutral atoms in a magnetic microchip trap. Coherence lifetimes exceeding 1 s are observed with atoms at distances of $4-130 \\mu$m from the microchip surface. The coherence lifetime in the microtrap is independent of atom-surface distance and agrees well with the results of similar measurements in macroscopic magnetic traps. Due to the absence of surface-induced decoherence, a miniaturized atomic clock with a relative stability in the $10^{-13}$ range can be realized. For applications in quantum information processing, we propose to use microwave near-fields in the proximity of chip wires to create potentials that depend on the internal state of the atoms.

  4. Customized oligonucleotide microchips that convert multiple genetic information to simple patterns, are portable and reusable

    Science.gov (United States)

    Mirzabekov, Andrei; Guschin, Dmitry Y.; Chik, Valentine; Drobyshev, Aleksei; Fotin, Alexander; Yershov, Gennadiy; Lysov, Yuri

    2002-01-01

    This invention relates to using customized oligonucleotide microchips as biosensors for the detection and identification of nucleic acids specific for different genes, organisms and/or individuals in the environment, in food and in biological samples. The microchips are designed to convert multiple bits of genetic information into simpler patterns of signals that are interpreted as a unit. Because of an improved method of hybridizing oligonucleotides from samples to microchips, microchips are reusable and transportable. For field study, portable laser or bar code scanners are suitable.

  5. Microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse-transcription polymerase chain reaction for the rapid detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens.

    Science.gov (United States)

    Jia, Ruan; Chengjun, Sun; Heng, Chen; Chen, Zhou; Yuanqian, Li; Yongxin, Li

    2015-07-01

    Enterovirus 71 and Coxsackievirus A16 are the main pathogens causing hand-foot-mouth disease. In this paper, microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse transcript-polymerase chain reaction has been developed for the detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens. The specific reverse transcription-polymerase chain reaction amplicons labeled with SYBR Orange were separated by microchip capillary electrophoresis and detected by laser induced fluorescence detector within 7 min. The intraday and interday relative standard deviation of migration time for DNA Marker was in the range of 1.36-2.94 and 2.78-3.96%, respectively. The detection limits were as low as 2.06 × 10(3) copies/mL for Enterovirus 71 and 5 × 10(3) copies/mL for Coxsackievirus A16. No cross-reactivity was observed with rotavirus, astrovirus, norovirus, and adenovirus, which showed good specificity of the method. This assay was validated using 100 throat swab specimens that were detected by real-time reverse-transcript polymerase chain reaction in parallel and the two methods produced the same results. This study provided a rapid, sensitive and specific method for the detection of Enterovirus 71 and Coxsackievirus A16, which make a contribution to significant time and cost saving for the identification and treatment of patients.

  6. Microchip atmospheric pressure chemical ionization source for mass spectrometry.

    Science.gov (United States)

    Ostman, Pekka; Marttila, Seppo J; Kotiaho, Tapio; Franssila, Sami; Kostiainen, Risto

    2004-11-15

    A novel microchip heated nebulizer for atmospheric pressure chemical ionization mass spectrometry is presented. Anisotropic wet etching is used to fabricate the flow channels, inlet, and nozzle on a silicon wafer. An integrated heater of aluminum is sputtered on a glass wafer. The two wafers are jointed by anodic bonding, creating a two-dimensional version of an APCI source with a sample channel in the middle and gas channels symmetrically on both sides. The ionization is initiated with an external corona-discharge needle positioned 2 mm in front of the microchip heated nebulizer. The microchip APCI source provides flow rates down to 50 nL/min, stable long-term analysis with chip lifetime of weeks, good quantitative repeatability (RSD 0.995) with linear dynamic rage of at least 4 orders of magnitude, and cost-efficient manufacturing. The limit of detection (LOD) for acridine measured with microchip APCI at flow rate of 6.2 muL/min was 5 nM, corresponding to a mass flow of 0.52 fmol/s. The LOD with commercial macro-APCI at a flow rate of 1 mL/min for acridine was the same, 5 nM, corresponding to a significantly worse mass flow sensitivity (83 fmol/s) than measured with microchip APCI. The advantages of microchip APCI makes it a very attractive new microfluidic detector.

  7. An overview of the use of microchips in electrophoretic separation techniques: fabrication, separation modes, sample preparation opportunities, and on-chip detection.

    Science.gov (United States)

    Hendrickx, Stijn; de Malsche, Wim; Cabooter, Deirdre

    2015-01-01

    This chapter is intended as a basic introduction to microchip-based capillary electrophoresis to set the scene for newcomers and give pointers to reference material. An outline of some commonly used setups and key concepts is given, many of which are explored in greater depth in later chapters.

  8. Potential Applications of Polymer Microchips in Pharmaceuticals

    Institute of Scientific and Technical Information of China (English)

    James J. Bao

    2000-01-01

    Some of the most recent development in polymer based microchips have been reviewed. Different methods used in fabricating ploymer microchips are introduced. The inportance and applications of the ploymer microchip in pharmaceutical and other areas are discussed. Finally, the potential problems and development areas in polymer microchip technology are also addressed.

  9. Microchips for CE: breakthroughs in real-world food analysis.

    Science.gov (United States)

    Escarpa, Alberto; González, María Cristina; López Gil, Miguel Angel; Crevillén, Agustín G; Hervás, Miriam; García, Miguel

    2008-12-01

    The well-known complexity of food matrices is approached using CE microchips with different strategies to improve the selectivity and sensitivity of the analysis by avoiding and/or making the sample preparation as simple as possible: (i) enhancing the peak capacity in order to perform direct injection, (ii) using the microchip platform to measure one target analyte/group of analytes with or without separating other related interferences, (iii) integrating sample preparation steps on the microchip platform, and (iv) integrating new analytical tools from nanotechnology in the detection stage. New analyte separations of food significance involving DNA probes, biogenic amines, vanilla flavors, and dyes have been reported as successfully breaking new barriers in areas of high impact in the market, such as transgenic food analysis, as well as the detection of frauds and toxins. Simple microchip layouts are still the most common designs used, though sophisticated new ones are emerging. In contrast to other application areas, electrochemical detection continues to be the most common detection route, followed by LIF, though non-conventional detection routes are also emerging, such as chemiluminescence or UV. In terms of analytical performance, the integration of calibration and quality control on a microchip platform, and remarkable accuracy and precision are being obtained using creative analytical methodologies that enhance the analytical potency of microfluidic chips for their future commercialization. This review critically states the most important advances derived from work done in the field over the past 2-3 years.

  10. In-channel indirect amperometric detection of nonelectroactive anions for electrophoresis on a poly(dimethylsiloxane) microchip.

    Science.gov (United States)

    Xu, Jing-Juan; Peng, Ying; Bao, Ning; Xia, Xing-Hua; Chen, Hong-Yuan

    2005-10-01

    In the present paper, we describe a microfluidics-based sensing system for nonelectroactive anions under negative separation electric field by mounting a single carbon fiber disk working electrode (WE) in the end part of a poly(dimethylsiloxane) microchannel. In contrast to work in a positive separation electric field described in our previous paper (Anal. Chem. 2004, 76, 6902-6907), here the electrochemical reduction reaction at the WE is not coupled with the separation high-voltage (HV) system, whereas the electrochemical oxidation reaction at the WE is coupled with the separation HV system. The electroactive indicator is the carbon fiber WE itself but not dissolved oxygen. This provides a convenient and sensitive means for the determination of nonelectroactive anions by amperometry. The influences of separation voltage, detection potential, and the distance between the WE and the separation channel outlet on the response of the detector have been investigated. The present detection mode is successfully used to electrochemically detect F-, Cl-, SO4(2-), CH3COO-, H2PO4-. Based on the preliminary results, a detection limit of 2 microM and a dynamic range up to three orders of magnitude for Cl- could be achieved.

  11. CE microchips: an opened gate to food analysis.

    Science.gov (United States)

    Escarpa, Alberto; González, María Cristina; Crevillén, Agustín González; Blasco, Antonio Javier

    2007-03-01

    CE microchips are the first generation of micrototal analysis systems (-TAS) emerging in the miniaturization scene of food analysis. CE microchips for food analysis are fabricated in both glass and polymer materials, such as PDMS and poly(methyl methacrylate) (PMMA), and use simple layouts of simple and double T crosses. Nowadays, the detection route preferred is electrochemical in both, amperometry and conductivity modes, using end-channel and contactless configurations, respectively. Food applications using CE microchips are now emerging since food samples present complex matrices, the selectivity being a very important challenge because the total integration of analytical steps into microchip format is very difficult. As a consequence, the first contributions that have recently appeared in the relevant literature are based primarily on fast separations of analytes of high food significance. These protocols are combined with different strategies to achieve selectivity using a suitable nonextensive sample preparation and/or strategically choosing detection routes. Polyphenolic compounds, amino acids, preservatives, and organic and inorganic ions have been studied using CE microchips. Thus, new and exciting future expectations arise in the domain of food analysis. However, several drawbacks could easily be found and assumed within the miniaturization map.

  12. Analysis of anions in ambient aerosols by microchip capillary electrophoresis.

    Science.gov (United States)

    Liu, Yan; MacDonald, David A; Yu, Xiao-Ying; Hering, Susanne V; Collett, Jeffrey L; Henry, Charles S

    2006-11-01

    We describe a microchip capillary electrophoresis method for the analysis of nitrate and sulfate in ambient aerosols. Investigating the chemical composition of ambient aerosol particles is essential for understanding their sources and effects. Significant progress has been made towards developing mass spectrometry-based instrumentation for rapid qualitative analysis of aerosols. Alternative methods for rapid quantification of selected high abundance compounds are needed to augment the capacity for widespread routine analysis. Such methods could provide much higher temporal and spatial resolution than can be achieved currently. Inorganic anions comprise a large percentage of particulate mass, with nitrate and sulfate among the most abundant species. While ion chromatography has proven very useful for analyzing extracts of time-integrated ambient aerosol samples collected on filters and for semi-continuous, on-line particle composition measurements, there is a growing need for development of new compact, inexpensive approaches to routine on-line aerosol ion analysis for deployment in spatially dense, atmospheric measurement networks. Microchip capillary electrophoresis provides the necessary speed and portability to address this need. In this report, on-column contact conductivity detection is used with hydrodynamic injection to create a simple microchip instrument for analysis of nitrate and sulfate. On-column contact conductivity detection was achieved using a Pd decoupler placed upstream from the working electrodes. Microchips containing two Au or Pd working electrodes showed a good linear range (5-500 microM) and low limits-of-detection for sulfate and nitrate, with Au providing the lowest detection limits (1 microM) for both ions. The completed microchip system was used to analyze ambient aerosol filter samples. Nitrate and sulfate concentrations measured by the microchip matched the concentrations measured by ion chromatography.

  13. Rapid Isolation and Detection for RNA Biomarkers for TBI Diagnostics

    Science.gov (United States)

    2016-10-01

    detection is very important for enabling future “liquid biopsy ” molecular diagnostics. 4.3 Impact on technology transfer The results of...transfer as it demonstrates overall viability of the DEP technology for a wide variety of diagnostic applications, including liquid biopsy cancer...Distribution Unlimited 13. SUPPLEMENTARY NOTES 14. ABSTRACT Our project work is focused on using a new dielectrophoresis (DEP) microarray technology for

  14. Analysis of proteins and peptides by electromigration methods in microchips.

    Science.gov (United States)

    Štěpánová, Sille; Kašička, Václav

    2017-01-01

    This review presents the developments and applications of microchip electromigration methods in the separation and analysis of peptides and proteins in the period 2011-mid-2016. The developments in sample preparation and preconcentration, microchannel material, and surface treatment are described. Separations by various microchip electromigration methods (zone electrophoresis in free and sieving media, affinity electrophoresis, isotachophoresis, isoelectric focusing, electrokinetic chromatography, and electrochromatography) are demonstrated. Advances in detection methods are reported and novel applications in the areas of proteomics and peptidomics, quality control of peptide and protein pharmaceuticals, analysis of proteins and peptides in biomatrices, and determination of physicochemical parameters are shown.

  15. Integrated Micro-Chip Amino Acid Chirality Detector for MOD

    Science.gov (United States)

    Glavin, D. P.; Bada, J. L.; Botta, O.; Kminek, G.; Grunthaner, F.; Mathies, R.

    2001-01-01

    Integration of a micro-chip capillary electrophoresis analyzer with a sublimation-based extraction technique, as used in the Mars Organic Detector (MOD), for the in-situ detection of amino acids and their enantiomers on solar system bodies. Additional information is contained in the original extended abstract.

  16. Neutral polymers as coatings for high resolution electrophoretic separation of Aβ peptides on glass microchips.

    Science.gov (United States)

    Mesbah, Kiarach; Verpillot, Romain; Chiari, Marcella; Pallandre, Antoine; Taverna, Myriam

    2014-12-21

    This study reports a comparison of the performances of two neutral polymers, poly ethylene-oxide (PEO) and poly(dimethylacrylamide-co-allyl glycidyl ether) (EpDMA), in glass microchips to achieve zone electrophoresis separation of several truncated forms of beta amyloid (Aβ) peptides, sharing very similar structures. The peptides were derivatized by FluoProbes 488 NHS to allow their fluorescence detection. Two protocols based either on PEO or EpDMA led to good pH stabilities in addition to a significant reduction of the electroosmotic flow. These two polymer coatings allowed repeatable analyses and high resolution for the simultaneous analysis of three Aβ peptides, Aβ 1-38, Aβ 1-40 and Aβ 1-42, considered as potential biomarkers of Alzheimer's disease. A recovery study showed that EpDMA was superior in reducing the adsorption of the Aβ peptides on the coated inner wall. Finally, the separation method relying on the EpDMA coated microchips was validated as linear using a calibration curve and the LOD was estimated to be close to 200 nM. Despite very short migration distances, different N-terminal or C-terminal truncated Aβ peptides, corresponding to promising biomarker combinations for the future diagnostic, were fully resolved. The method was successfully applied to detect these peptides in spiked cerebrospinal fluid and has provided a first achievement towards the development of a microsystem that would integrate preconcentration and separation steps.

  17. Dynamics of and diagnostic methods for detecting small carious lesions

    NARCIS (Netherlands)

    Pine, CM; tenBosch, JJ

    1996-01-01

    This paper reviews and discusses the dynamics of carious lesions and diagnostic methods for their detection. Summary data are presented on progression rates of differing lesion types with differing initial severity. Among the non-invasive techniques, fibre-optic transillumination, used appropriately

  18. Standardization of diagnostic PCR for the detection of foodborne pathogens

    DEFF Research Database (Denmark)

    Malorny, B.; Tassios, P.T.; Radstrom, P.

    2003-01-01

    In vitro amplification of nucleic acids using the polymerase chain reaction (PCR) has become, since its discovery in the 1980s, a powerful diagnostic tool for the analysis of microbial infections as well as for the analysis of microorganisms in food samples. However, despite its potential, PCR has...... neither gained wide acceptance in routine diagnostics nor been widely incorporated in standardized methods. Lack of validation and standard protocols, as well as variable quality of reagents and equipment, influence the efficient dissemination of PCR methodology from expert research laboratories to end......-user laboratories. Moreover, the food industry understandably requires and expects officially approved standards. Recognizing this, in 1999, the European Commission approved the research project, FOOD-PCR (http://www.PCR.dk), which aims to validate and standardize the use of diagnostic PCR for the detection...

  19. The Diagnostic Value of Gastrin-17 Detection in Atrophic Gastritis

    Science.gov (United States)

    Wang, Xu; Ling, Li; Li, Shanshan; Qin, Guiping; Cui, Wei; Li, Xiang; Ni, Hong

    2016-01-01

    Abstract A meta-analysis was performed to assess the diagnostic value of gastrin-17 (G-17) for the early detection of chronic atrophic gastritis (CAG). An extensive literature search was performed, with the aim of selecting publications that reported the accuracy of G-17 in predicting CAG, in the following databases: PubMed, Science Direct, Web of Science, Chinese Biological Medicine, Chinese National Knowledge Infrastructure, Wanfang, and VIP. To assess the diagnostic value of G-17, the following statistics were estimated and described: sensitivity, specificity, diagnostic odds ratios (DOR), summary receiver operating characteristic curves, area under the curve (AUC), and 95% confidence intervals (CIs). Thirteen studies that met the inclusion criteria were included in this meta-analysis, comprising 894 patients and 1950 controls. The pooled sensitivity and specificity of these studies were 0.48 (95% CI: 0.45–0.51) and 0.79 (95% CI: 0.77–0.81), respectively. The DOR was 5.93 (95% CI: 2.93–11.99), and the AUC was 0.82. G-17 may have potential diagnostic value because it has good specificity and a moderate DOR and AUC for CAG. However, more studies are needed to improve the sensitivity of this diagnostic tool in the future. PMID:27149493

  20. Fabrication of PMMA CE microchips by infrared-assisted polymerization.

    Science.gov (United States)

    Chen, Yun; Duan, Haotian; Zhang, Luyan; Chen, Gang

    2008-12-01

    In this report, a method based on the infrared-assisted polymerization of methyl methacrylate has been developed for the rapid fabrication of PMMA CE microchips. Methyl methacrylate containing AIBN was allowed to prepolymerize in a water bath to form a fast-curing molding solution that was subsequently sandwiched between a silicon template and a piece of 1 mm-thick PMMA plate. The images of microchannels on the silicon template were precisely replicated into the synthesized PMMA substrates during the infrared-assisted polymerization of the molding solution. The polymerization could be completed within 50 min at 50 degrees C. The obtained channel plate was subsequently bonded to a piece of PMMA cover sheet to form a microchip with the aid of heat and pressure. The new fabrication approach obviates the need for special equipment and significantly simplifies the process of fabricating PMMA microchips. The attractive performance of the obtained PMMA microchips has been demonstrated in connection with contactless conductivity detection for the separation and detection of ionic species.

  1. Diagnostic trends in Clostridium difficile detection in Finnish microbiology laboratories.

    Science.gov (United States)

    Könönen, Eija; Rasinperä, Marja; Virolainen, Anni; Mentula, Silja; Lyytikäinen, Outi

    2009-12-01

    Due to increased interest directed to Clostridium difficile-associated infections, a questionnaire survey of laboratory diagnostics of toxin-producing C. difficile was conducted in Finland in June 2006. Different aspects pertaining to C. difficile diagnosis, such as requests and criteria used for testing, methods used for its detection, yearly changes in diagnostics since 1996, and the total number of investigations positive for C. difficile in 2005, were asked in the questionnaire, which was sent to 32 clinical microbiology laboratories, including all hospital-affiliated and the relevant private clinical microbiology laboratories in Finland. The situation was updated by phone and email correspondence in September 2008. In June 2006, 28 (88%) laboratories responded to the questionnaire survey; 24 of them reported routinely testing requested stool specimens for C. difficile. Main laboratory methods included toxin detection (21/24; 88%) and/or anaerobic culture (19/24; 79%). In June 2006, 18 (86%) of the 21 laboratories detecting toxins directly from feces, from the isolate, or both used methods for both toxin A (TcdA) and B (TcdB), whereas only one laboratory did so in 1996. By September 2008, all of the 23 laboratories performing diagnostics for C. difficile used methods for both TcdA and TcdB. In 2006, the number of specimens processed per 100,000 population varied remarkably between different hospital districts. In conclusion, culturing C. difficile is common and there has been a favorable shift in toxin detection practice in Finnish clinical microbiology laboratories. However, the variability in diagnostic activity reported in 2006 creates a challenge for national monitoring of the epidemiology of C. difficile and related diseases.

  2. Microchip device for liquid phase analysis

    Energy Technology Data Exchange (ETDEWEB)

    Ramsey, j.m.

    2000-05-01

    The lab-on-a-chip concept has enabled miniature instruments to be developed that allow the rapid execution and automation of fluidic operations such as valving, separation, dilution, mixing, and flow splitting upon the proper application of a motive (driving) force. The integration of these simple operations to perform complete, multiple-step chemical assays is rapidly becoming a reality. Such compact, monolithic devices potentially enjoy advantages in speed, cost, automation, reagent consumption, and waste generation compared to existing laboratory-scale instruments. Initial reports of these microfluidic devices focused on combining various electrokinetically driven separation methods including microchip electrophoresis, gel electrophoresis, micellar electrokinetic chromatography (MEKC) and open channel electrochromatography (OCEC) with fluidic valving to introduce sample plugs into the separation channel. Other operations have quickly been integrated with the separations and fluidic valving on these microchips. For example, integrated devices with mixers/diluters for precolumn and postcolumn analyte derivatization, deoxyribonucleic acid (DNA) restriction digests, enzyme assays, and polymerase chain reaction (PCR) amplification have been added to the basic design. Integrated mixers that can perform solvent programming for both MEKC and OCEC have also been demonstrated. These examples are simple, yet powerful, demonstrations of the potential for lab-on-a-chip devices. In this report, three key areas for improved performance of these devices are described: on-chip calibration techniques, enhanced separative performance, and enhanced detection capabilities.

  3. Center conductor diagnostic for multipactor detection in inaccessible geometries

    Science.gov (United States)

    Chaplin, Vernon H.; Hubble, Aimee A.; Clements, Kathryn A.; Graves, Timothy P.

    2017-01-01

    Electron collecting current probes are the most reliable diagnostic of multipactor and radiofrequency (RF) ionization breakdown; however, stand-alone probes can only be used in test setups where the breakdown region is physically accessible. This paper describes techniques for measuring multipactor current directly on the center conductor of a coaxial RF device (or more generally, on the signal line in any two-conductor RF system) enabling global multipactor detection with improved sensitivity compared to other common diagnostics such as phase null, third harmonic, and reflected power. The center conductor diagnostic may be AC coupled for use in systems with a low DC impedance between the center conductor and ground. The effect of DC bias on the breakdown threshold was studied: in coaxial geometry, the change in threshold was positive biases satisfying VD C/VR F 0 effectively with no bias at all—this is the preferred implementation, but biases in the range VD C=0 -10 V may be applied if necessary. The polarity of the detected current signal may be positive or negative depending on whether there is net electron collection or emission globally.

  4. Quantum dots in diagnostics and detection: principles and paradigms.

    Science.gov (United States)

    Pisanic, T R; Zhang, Y; Wang, T H

    2014-06-21

    Quantum dots are semiconductor nanocrystals that exhibit exceptional optical and electrical behaviors not found in their bulk counterparts. Following seminal work in the development of water-soluble quantum dots in the late 1990's, researchers have sought to develop interesting and novel ways of exploiting the extraordinary properties of quantum dots for biomedical applications. Since that time, over 10,000 articles have been published related to the use of quantum dots in biomedicine, many of which regard their use in detection and diagnostic bioassays. This review presents a didactic overview of fundamental physical phenomena associated with quantum dots and paradigm examples of how these phenomena can and have been readily exploited for manifold uses in nanobiotechnology with a specific focus on their implementation in in vitro diagnostic assays and biodetection.

  5. Rapid amplification of genetically modified organisms using a circular ferrofluid-driven PCR microchip.

    Science.gov (United States)

    Sun, Yi; Kwok, Yien-Chian; Foo-Peng Lee, Peter; Nguyen, Nam-Trung

    2009-07-01

    The use of genetically modified organisms (GMOs) as food and in food products is becoming more and more widespread. Polymerase chain reaction (PCR) technology is extensively used for the detection of GMOs in food products in order to verify compliance with labeling requirements. In this paper, we present a novel close-loop ferrofluid-driven PCR microchip for rapid amplification of GMOs. The microchip was fabricated in polymethyl methacrylate by CO2 laser ablation and was integrated with three temperature zones. PCR solution was contained in a circular closed microchannel and was driven by magnetic force generated by an external magnet through a small oil-based ferrofluid plug. Successful amplification of genetically modified soya and maize were achieved in less than 13 min. This PCR microchip combines advantages of cycling flexibility and quick temperature transitions associated with two existing microchip PCR techniques, and it provides a cost saving and less time-consuming way to conduct preliminary screening of GMOs.

  6. 78 FR 33842 - Determination and Declaration Regarding Emergency Use of In Vitro Diagnostics for Detection of...

    Science.gov (United States)

    2013-06-05

    ... Regarding Emergency Use of In Vitro Diagnostics for Detection of Middle East Respiratory Syndrome... vitro diagnostics for detection of Middle East respiratory syndrome coronavirus (MERS-CoV) pursuant to... the FDA, HHS, issue an EUA for in vitro diagnostics for detection of Middle East respiratory...

  7. Pyrosequencing for EGFR mutation detection: diagnostic accuracy and clinical implications.

    Science.gov (United States)

    Sahnane, Nora; Gueli, Rossana; Tibiletti, Maria G; Bernasconi, Barbara; Stefanoli, Michele; Franzi, Francesca; Pinotti, Graziella; Capella, Carlo; Furlan, Daniela

    2013-12-01

    EGFR-activating mutations predict responsiveness to EGFR tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) patients. Mutation screening is crucial to support therapeutic decisions and is commonly conducted using dideoxy sequencing, although its sensitivity is suboptimal in clinical settings. To evaluate the diagnostic performance of pyrosequencing and dideoxy sequencing, we examined EGFR mutation status in a retrospective cohort of 53 patients with NSCLCs clinically selected for TKI therapy and whose clinical outcome was available. Moreover, pyrosequencing quantitative results were compared with EGFR amplification data. EGFR mutations were investigated by pyrosequencing and by dideoxy sequencing. Detection rates of both methods were determined by titration assays using NCI-H1975 and HCC-827 cell lines. Increased EGFR copy number was assessed by fluorescence in situ hybridization (FISH). Pyrosequencing showed a higher detection rate than dideoxy sequencing. Tumor control rate of cases with mutant and wild-type EGFR was 86% and 29%, respectively. EGFR amplification was significantly associated with EGFR mutation and a positive correlation between high percentages of mutant alleles and clinical response to TKI was observed. We concluded that pyrosequencing is more sensitive than dideoxy sequencing in mutation screening for EGFR mutations. Detection rate of dideoxy sequencing was suboptimal when low frequencies of mutant alleles or low tumor cell contents were observed. Pyrosequencing enables quantification of mutant alleles that correlates well with increased EGFR copy number assessed by FISH. Pyrosequencing should be used in molecular diagnostic of NSCLC to appropriately select patients who are likely to benefit from TKI therapy.

  8. Current Status on Stress Diagnostic Kit and Detection Technology

    Energy Technology Data Exchange (ETDEWEB)

    Park, Sang Hyun; Choi, Mi Hee; Ko, Kyong Cheol

    2008-06-15

    The accurate measurement of a stress level is one of the most important issues in a stress diagnosis and its measurement could be of great value in clinical medicine. Stress has a potent effect on the spirit and physical condition of an individual. There are various methods available for its measurement. Some of the commonly used techniques for the diagnosis of a stress level include analysis of the body fluids, questionnaire assessments, psychophysiological evaluations and by determining heart rate variability (HRV) of subjects. However, the existing diagnostic methods have several defects like, a low sensitivity, inaccuracy and long of operation time. In this report, we present a diagnostic technology to detect a stress level which is the origin of various diseases. This method can be of great help in providing an early diagnosis through a biosensor and might play a vital role in preventing diseases like hypochondria and hypertension. Majority of the human population is exposed to stress in one way or another and hence developing a convenient stress diagnosis kit will be of great use to all. This stress diagnostic kit and detection technology dose not involve simple a mechanical measurement or questionnaires, but is based on developing a detection kit with a high sensitivity, which will mean an easy use for common man. Individuals can undergo regular check ups and can personally diagnose their present situation of health by determining their stress levels, thus enabling them to diagnose the early onset of several stress disorders. This might help them take precautionary measures and thereby lead to a healthy life.

  9. Computational diagnostics for detecting phase transitions during nanoindentation

    Energy Technology Data Exchange (ETDEWEB)

    Lee, S.M.; Hoover, C.G.; Kallman, J.S.; De Groot, A.J. (Lawrence Livermore National Lab., CA (United States)); Hoover, W.G. (Lawrence Livermore National Lab., CA (United States) California Univ., Livermore, CA (United States). Div. of Applied Science); Wooten, F. (California Univ., Livermore, CA (United States). Div. of Applied Science)

    1992-12-01

    We study nanoindenmtion of silicon using nonequilibrium molecular dynamics simulations. with up to a million particles. Both crystalline and amorphous silicon samples are considered. We use compumtional diffraction pattems as a diagnostic tool for detecting phase transitions resulting from structural changes. Simulations of crystalline samples show a transition to the amorphous phase in a region a few atomic layers thick surrounding the lateral faces of the indenter, as has been suggested by experimental results. Our simulation results provide estimates for the yield strength (nanohardness) of silicon for a range of temperatures.

  10. Development of a real-time microchip PCR system for portable plant disease diagnosis.

    Directory of Open Access Journals (Sweden)

    Chiwan Koo

    Full Text Available Rapid and accurate detection of plant pathogens in the field is crucial to prevent the proliferation of infected crops. Polymerase chain reaction (PCR process is the most reliable and accepted method for plant pathogen diagnosis, however current conventional PCR machines are not portable and require additional post-processing steps to detect the amplified DNA (amplicon of pathogens. Real-time PCR can directly quantify the amplicon during the DNA amplification without the need for post processing, thus more suitable for field operations, however still takes time and require large instruments that are costly and not portable. Microchip PCR systems have emerged in the past decade to miniaturize conventional PCR systems and to reduce operation time and cost. Real-time microchip PCR systems have also emerged, but unfortunately all reported portable real-time microchip PCR systems require various auxiliary instruments. Here we present a stand-alone real-time microchip PCR system composed of a PCR reaction chamber microchip with integrated thin-film heater, a compact fluorescence detector to detect amplified DNA, a microcontroller to control the entire thermocycling operation with data acquisition capability, and a battery. The entire system is 25 × 16 × 8 cm(3 in size and 843 g in weight. The disposable microchip requires only 8-µl sample volume and a single PCR run consumes 110 mAh of power. A DNA extraction protocol, notably without the use of liquid nitrogen, chemicals, and other large lab equipment, was developed for field operations. The developed real-time microchip PCR system and the DNA extraction protocol were used to successfully detect six different fungal and bacterial plant pathogens with 100% success rate to a detection limit of 5 ng/8 µl sample.

  11. Microchips in Medicine: Current and Future Applications

    OpenAIRE

    Eltorai, Adam E. M.; Henry Fox; Emily McGurrin; Stephanie Guang

    2016-01-01

    With the objective of improving efficacy and morbidity, device manufacturers incorporate chemicals or drugs into medical implants. Using multiple reservoirs of discrete drug doses, microchips represent a new technology capable of on-demand release of various drugs over long periods of time. Herein, we review drug delivery systems, how microchips work, recent investigations, and future applications in various fields of medicine.

  12. Production of Microchips from Polystyrene Plates

    Science.gov (United States)

    Pace, Sarah Lindsey

    2009-01-01

    Currently manufactured microchips are expensive to make, require specialized equipment, and leave a large environmental footprint. To counter this, an alternative procedure that is cheaper and leaves a smaller environmental footprint should be made. The goal of this research project is to develop a process that creates microchips from polystyrene…

  13. Microchips in Medicine: Current and Future Applications.

    Science.gov (United States)

    Eltorai, Adam E M; Fox, Henry; McGurrin, Emily; Guang, Stephanie

    2016-01-01

    With the objective of improving efficacy and morbidity, device manufacturers incorporate chemicals or drugs into medical implants. Using multiple reservoirs of discrete drug doses, microchips represent a new technology capable of on-demand release of various drugs over long periods of time. Herein, we review drug delivery systems, how microchips work, recent investigations, and future applications in various fields of medicine.

  14. [Development of microchips for the analysis of biomarkers in blood].

    Science.gov (United States)

    Kataoka, Masatoshi; Abe, Kaori; Hashimoto, Yoshiko; Yamamura, Shohei; Yatsushiro, Shouki

    2012-11-01

    Several types of microchips have been developed for application in clinical diagnosis. A microchip made of cyclic olefin copolymer with straight microchannels (300 microm width and 100 microm depth) was employed for sandwich ELISA for the determination of serum type I C-peptide (PICP), a biomarker of osteoporosis. This assay enabled us to determine PICP with accuracy and high sensitivity, reducing the time for the immunoassay to 1/6, and the consumption of samples and reagents to 1/50 compared with the conventional method. Furthermore, cell microarray chips with 20,944 microchambers (105 microm width and 50 microm depth), made of polystyrene, were employed for malaria diagnosis and the detection of carcinoma cells among the leukocytes. Around 100 erythrocytes or leukocytes were accommodated in each microchamber with the formation of a monolayer. For malaria diagnosis, it offered 10-100 times higher sensitivity in the detection of malaria infected erythrocytes than conventional light microscopy, and easy operation within 15 min. By double staining for epithelial cells on the cell microarray chip, one carcinoma cell could be detected among 1,800,000 leukocytes. These results indicate the potential of microchips for clinic diagnosis.

  15. 76 FR 28689 - Microbiology Devices; Classification of In Vitro Diagnostic Device for Bacillus Species Detection

    Science.gov (United States)

    2011-05-18

    ... devices for Bacillus spp. detection (Ref. 1). A. Identification FDA is proposing the following identification based on the Panel's recommendation and the available information. An in vitro diagnostic device... diagnostic device for Bacillus spp. detection. (a) Identification. An in vitro diagnostic device for Bacillus...

  16. Diamond electrophoretic microchips-Joule heating effects

    Energy Technology Data Exchange (ETDEWEB)

    Karczemska, Anna T., E-mail: anna.karczemska@p.lodz.pl [Technical University of Lodz, Institute of Turbomachinery, 219/223 Wolczanska str., Lodz (Poland); Witkowski, Dariusz [Technical University of Lodz, Institute of Turbomachinery, 219/223 Wolczanska str., Lodz (Poland); Ralchenko, Victor, E-mail: ralchenko@nsc.gpi.ru [General Physics Institute, Russian Academy of Science, 38 Vavilov str., Moscow (Russian Federation); Bolshakov, Andrey; Sovyk, Dmitry [General Physics Institute, Russian Academy of Science, 38 Vavilov str., Moscow (Russian Federation); Lysko, Jan M., E-mail: jmlysko@ite.waw.pl [Institute of Electron Technology, Al. Lotnikow 32/46, 02-668 Warsaw (Poland); Fijalkowski, Mateusz, E-mail: petr.louda@vslib.cz [Technical University of Liberec, Faculty of Mechanical Engineering (Czech Republic); Bodzenta, Jerzy, E-mail: jerzy.bodzenta@polsl.pl [Silesian University of Technology, Institute of Physics, 2 Krzywoustego str., 44-100 Gliwice (Poland); Hassard, John, E-mail: j.hassard@imperial.ac.uk [Imperial College of Science, Technology and Medicine, London (United Kingdom)

    2011-03-15

    Microchip electrophoresis (MCE) has become a mature separation technique in the recent years. In the presented research, a polycrystalline diamond electrophoretic microchip was manufactured with a microwave plasma chemical vapour deposition (MPCVD) method. A replica technique (mould method) was used to manufacture microstructures in diamond. A numerical analysis with CoventorWare{sup TM} was used to compare thermal properties during chip electrophoresis of diamond and glass microchips of the same geometries. Temperature distributions in microchips were demonstrated. Thermal, electrical, optical, chemical and mechanical parameters of the polycrystalline diamond layers are advantageous over traditionally used materials for microfluidic devices. Especially, a very high thermal conductivity coefficient gives a possibility of very efficient dissipation of Joule heat from the diamond electrophoretic microchip. This enables manufacturing of a new generation of microdevices.

  17. MODERN DIAGNOSTIC TOOLS FOR DETECTING HEPATIC FIBROSIS IN CHILDREN

    Directory of Open Access Journals (Sweden)

    A.N. Surkov

    2009-01-01

    Full Text Available The present review highlights the new diagnostic tools for detecting hepatic fibrosis against the backdrop of different liver pathologies in children. They are characterized by a variety of the clinical forms, progressive course along with the fibroid changes in liver and possible result in the liver cirrhosis. The authors considered the modern methods of the needle liver biopsy and noninvasive visualization, as well as justified the necessity to look for the noninvasive markers of the fibroid liver by means of the antibody'mediated diagnostic methods. Such tests are based on identification of various molecular compounds, which are fibrogenesis activators and take part in the formation of the extracellular matrix components. They also described changes of such fibroid liver markers, as hyaluronic acid, collagen type IV, matrix metalloproteinases 2 and 9, tissular inhibitor of the matrix metalloproteinases'1, transforming growth factor–1, which may be recommended for the noninvasive monitoring of the hepatic fibrosis in children.Key words: hepatic fibrosis, antibody'mediated fibrosis markers, children.

  18. Multiplexed electrochemical protein detection and translation to personalized cancer diagnostics.

    Science.gov (United States)

    Rusling, James F

    2013-06-04

    Measuring diagnostic panels of multiple proteins promises a new, personalized approach to early detection and therapy of diseases like cancer. Levels of biomarker proteins in patient serum can provide a continually updated record of disease status. Research in electrochemical detection of proteins has produced exquisitely sensitive approaches. Most utilize ELISA-like sandwich immunoassays incorporating various aspects of nanotechnology. Several of these ultrasensitive methodologies have been extended to microfluidic multiplexed protein detection, but engineered solutions are needed to measure more proteins in a single device from a small patient sample such as a drop of blood or tissue lysate. To achieve clinical or point-of-care (POC) use, simplicity and low cost are essential. In multiplexed microfluidic immunoassays, required reagent additions and washing steps pose a significant problem calling for creative engineering. A grand challenge is to develop a general cancer screening device to accurately measure 50-100 proteins in a simple, cost-effective fashion. This will require creative solutions to simplified reagent addition and multiplexing.

  19. Millimeter Wave Spectroscopy for Breast Cancer Diagnostics and Detection

    Science.gov (United States)

    Korolev, Konstantin; Chen, Shu; Afsar, Mohammed; Naber, Stephen

    2009-03-01

    Broad-band millimeter wave transmittance measurements of normal and tumorous (cancerous) human breast tissue samples have been acquired in--vitro by employing a free-space, quasi-optical spectrometer. Freshly excised breast tissues were prepared and preserved in 10% neutral-buffered formalin solution before testing. Significant differences in the transmittance profiles have been found between the normal and tumorous tissues. It has been found that despite the inhomogeneity and variable structure and composition of each single tissue, the tumorous specimens consistently manifest much higher absorption level of millimeter wave radiation than the normal ones. It has been shown that free space, quasi-optical spectrometer is capable of contributing valuable insights into the dielectric properties of normal and tumorous human breast tissues and aiding in further developments of millimeter wave spectroscopy and mammography for the breast cancer diagnostics and detection.

  20. [THE POSSIBILITIES OF APPLICATION OF TECHNOLOGY PROTEIN MICROARRAY (MICROCHIPS) FOR ANALYSIS OF PROTEIN COMPOSITION OF BLOOD SERUM].

    Science.gov (United States)

    Gumanova, N G; Klimushina, M V; Metelskaya, V A; Boitsov, S A

    2015-10-01

    The microchip technology represents convenient and relatively economic tool of analyzing specific biomarkers with the purpose to diagnose diseases, to evaluate effectiveness of therapy and to investigate signaling pathways. To analyze protein composition of blood serum certain types of finished microchips which were not applied previously on the territory of Russia. The detection from 2% to 5% out of matrix of chips depending on their variety was managed without preliminary depletion of serum (removal of proteins of major fractions). Hence, partial protein composition of blood serum can be analyzed with microchips even without preliminary removal of proteins of major fractions.

  1. Simple filter microchip for rapid separation of plasma and viruses from whole blood

    Directory of Open Access Journals (Sweden)

    Wang SQ

    2012-09-01

    Full Text Available ShuQi Wang,1 Dusan Sarenac,1 Michael H Chen,1 Shih-Han Huang,1 Francoise F Giguel,2 Daniel R Kuritzkes,3 Utkan Demirci1,41Bio-acoustic MEMS in Medicine Laboratory, Department of Medicine, Division of Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA; 2Infectious Diseases Unit, Massachusetts General Hospital, Boston, MA, USA; 3Section of Retroviral Therapeutics, Brigham and Women's Hospital, Boston, MA, USA; 4Harvard-MIT Health Sciences and Technology, Cambridge, MA, USAAbstract: Sample preparation is a significant challenge for detection and sensing technologies, since the presence of blood cells can interfere with the accuracy and reliability of virus detection at the nanoscale for point-of-care testing. To the best of our knowledge, there is not an existing on-chip virus isolation technology that does not use complex fluidic pumps. Here, we presented a lab-on-a-chip filter device to isolate plasma and viruses from unprocessed whole blood based on size exclusion without using a micropump. We demonstrated that viruses (eg, HIV can be separated on a filter-based chip (2-µm pore size from HIV-spiked whole blood at high recovery efficiencies of 89.9% ± 5.0%, 80.5% ± 4.3%, and 78.2% ± 3.8%, for viral loads of 1000, 10,000 and 100,000 copies/mL, respectively. Meanwhile, 81.7% ± 6.7% of red blood cells and 89.5% ± 2.4% of white blood cells were retained on 2 µm pore–sized filter microchips. We also tested these filter microchips with seven HIV-infected patient samples and observed recovery efficiencies ranging from 73.1% ± 8.3% to 82.5% ± 4.1%. These results are first steps towards developing disposable point-of-care diagnostics and monitoring devices for resource-constrained settings, as well as hospital and primary care settings.Keywords: microchip, filtration, virus isolation, plasma separation, point-of-care

  2. Bioinformatics for Diagnostics, Forensics, and Virulence Characterization and Detection

    Energy Technology Data Exchange (ETDEWEB)

    Gardner, S; Slezak, T

    2005-04-05

    We summarize four of our group's high-risk/high-payoff research projects funded by the Intelligence Technology Innovation Center (ITIC) in conjunction with our DHS-funded pathogen informatics activities. These are (1) quantitative assessment of genomic sequencing needs to predict high quality DNA and protein signatures for detection, and comparison of draft versus finished sequences for diagnostic signature prediction; (2) development of forensic software to identify SNP and PCR-RFLP variations from a large number of viral pathogen sequences and optimization of the selection of markers for maximum discrimination of those sequences; (3) prediction of signatures for the detection of virulence, antibiotic resistance, and toxin genes and genetic engineering markers in bacteria; (4) bioinformatic characterization of virulence factors to rapidly screen genomic data for potential genes with similar functions and to elucidate potential health threats in novel organisms. The results of (1) are being used by policy makers to set national sequencing priorities. Analyses from (2) are being used in collaborations with the CDC to genotype and characterize many variola strains, and reports from these collaborations have been made to the President. We also determined SNPs for serotype and strain discrimination of 126 foot and mouth disease virus (FMDV) genomes. For (3), currently >1000 probes have been predicted for the specific detection of >4000 virulence, antibiotic resistance, and genetic engineering vector sequences, and we expect to complete the bioinformatic design of a comprehensive ''virulence detection chip'' by August 2005. Results of (4) will be a system to rapidly predict potential virulence pathways and phenotypes in organisms based on their genomic sequences.

  3. 78 FR 25273 - Determination and Declaration Regarding Emergency Use of in Vitro Diagnostics for Detection of...

    Science.gov (United States)

    2013-04-30

    ... HUMAN SERVICES Determination and Declaration Regarding Emergency Use of in Vitro Diagnostics for... authorization of emergency use of in vitro diagnostics for detection of the avian influenza A (H7N9) virus... an emergency with respect to in vitro diagnostics as defined under the Public Readiness and...

  4. Capillary liquid chromatography-microchip atmospheric pressure chemical ionization-mass spectrometry.

    Science.gov (United States)

    Ostman, Pekka; Jäntti, Sirkku; Grigoras, Kestas; Saarela, Ville; Ketola, Raimo A; Franssila, Sami; Kotiaho, Tapio; Kostiainen, Risto

    2006-07-01

    A miniaturized nebulizer chip for capillary liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (capillary LC-microchip APCI-MS) is presented. The APCI chip consists of two wafers, a silicon wafer and a Pyrex glass wafer. The silicon wafer has a DRIE etched through-wafer nebulizer gas inlet, an edge capillary insertion channel, a stopper, a vaporizer channel and a nozzle. The platinum heater electrode and pads for electrical connection were patterned on to the Pyrex glass wafer. The two wafers were joined by anodic bonding, creating a microchip version of an APCI-source. The sample inlet capillary from an LC column is directly connected to the vaporizer channel of the APCI chip. The etched nozzle in the microchip forms a narrow sample plume, which is ionized by an external corona needle, and the formed ions are analyzed by a mass spectrometer. The nebulizer chip enables for the first time the use of low flow rate separation techniques with APCI-MS. The performance of capillary LC-microchip APCI-MS was tested with selected neurosteroids. The capillary LC-microchip APCI-MS provides quantitative repeatability and good linearity. The limits of detection (LOD) with a signal-to-noise ratio (S/N) of 3 in MS/MS mode for the selected neurosteroids were 20-1000 fmol (10-500 nmol l(-1)). LODs (S/N = 3) with commercial macro APCI with the same compounds using the same MS were about 10 times higher. Fast heat transfer allows the use of the optimized temperature for each compound during an LC run. The microchip APCI-source provides a convenient and easy method to combine capillary LC to any API-MS equipped with an APCI source. The advantages and potentials of the microchip APCI also make it a very attractive interface in microfluidic APCI-MS.

  5. Diagnostic Accuracy of Ultrasound in Detection of Traumatic Lens Dislocation

    Directory of Open Access Journals (Sweden)

    Seyed Hossein Ojaghi Haghighi

    2014-08-01

    Full Text Available Introduction: Traumatic eye injuries (TEI involved about 3% of cases referred to the emergency departments of developing countries. Lens dislocation is one of the critical cases of ophthalmic emergencies. The present study was aimed to evaluate the diagnostic accuracy of ultrasonography in detection of traumatic lens dislocation. Methods: In this cross-sectional study the findings of ultrasonography and orbital computed tomography (OCT of head and face trauma patients, referred to Imam Reza hospital, Tabriz, Iran, from July 2013 to June 2014, have been compared. The sensitivity, specificity, positive and negative likelihood ratio, positive and negative predictive value, and accuracy of ultrasonography were calculated. Cohen's kappa coefficient was presented to assess the agreement of ultrasonography with OCT findings. Results: One hundred thirty patients with the mean age of 35.4±18.0 were evaluated (75.4% male. Sensitivity and specificity of ultrasonography were 84.6% (95% Cl: 53.7-97.3 and 98.3% (95% Cl: 93.3- 99.7, respectively. Also, positive and negative likelihood ratio were calculated 49.5 (95% Cl: 12.3-199.4 and 0.15 (95% Cl: 0.04- 0.56, respectively. Cohen's kappa coefficient of 0.83 (95% Cl: 0.66-1.0; p<0.0001 was representative of excellent agreement of these two tests. Conclusion: The finding of this project was representative of 84.6% sensitivity, 98.3% specificity, and 96.9% accuracy of ultrasonography in detection of traumatic lens dislocation. It seems that in cases which OCT is not possible, ultrasonography could be an acceptable option to assess traumatic eye injuries.

  6. Microchips in Medicine: Current and Future Applications

    Directory of Open Access Journals (Sweden)

    Adam E. M. Eltorai

    2016-01-01

    Full Text Available With the objective of improving efficacy and morbidity, device manufacturers incorporate chemicals or drugs into medical implants. Using multiple reservoirs of discrete drug doses, microchips represent a new technology capable of on-demand release of various drugs over long periods of time. Herein, we review drug delivery systems, how microchips work, recent investigations, and future applications in various fields of medicine.

  7. Diagnostic values for skin temperature assessment to detect diabetes-related foot complications

    NARCIS (Netherlands)

    Netten, van Jaap J.; Prijs, Miranda; Baal, van Jeff G.; Liu, Chanjuan; Heijden, van der Ferdi; Bus, Sicco A.

    2014-01-01

    Skin temperature assessment is a promising modality for early detection of diabetic foot problems, but its diagnostic value has not been studied. Our aims were to investigate the diagnostic value of different cutoff skin temperature values for detecting diabetes-related foot complications such as ul

  8. A Peristaltic Pump Integrated on a 100% Glass Microchip Using Computer Controlled Piezoelectric Actuators

    Directory of Open Access Journals (Sweden)

    Yo Tanaka

    2014-05-01

    Full Text Available Lab-on-a-chip technology is promising for the miniaturization of chemistry, biochemistry, and/or biology researchers looking to exploit the advantages of a microspace. To manipulate fluid on a microchip, on-chip pumps are indispensable. To date, there have been several types of on-chip pumps including pneumatic, electroactive, and magnetically driven. However these pumps introduce polymers, metals, and/or silicon to the microchip, and these materials have several disadvantages, including chemical or physical instability, or an inherent optical detection limit. To overcome/avoid these issues, glass has been one of the most commonly utilized materials for the production of multi-purpose integrated chemical systems. However, glass is very rigid, and it is difficult to incorporate pumps onto glass microchips. This paper reports the use of a very flexible, ultra-thin glass sheet (minimum thickness of a few micrometers to realize a pump installed on an entirely glass-based microchip. The pump is a peristaltic-type, composed of four serial valves sealing a cavity with two penetrate holes using ultra-thin glass sheet. By this pump, an on-chip circulating flow was demonstrated by directly observing fluid flow, visualized via polystyrene tracking particles. The flow rate was proportional to the pumping frequency, with a maximum flow rate of approximately 0.80 μL/min. This on-chip pump could likely be utilized in a wide range of applications which require the stability of a glass microchip.

  9. Gas chromatography-microchip atmospheric pressure chemical ionization-mass spectrometry.

    Science.gov (United States)

    Ostman, Pekka; Luosujärvi, Laura; Haapala, Markus; Grigoras, Kestas; Ketola, Raimo A; Kotiaho, Tapio; Franssila, Sami; Kostiainen, Risto

    2006-05-01

    An atmospheric pressure chemical ionization (APCI) microchip is presented for combining a gas chromatograph (GC) to a mass spectrometer (MS). The chip includes capillary insertion channel, stopper, vaporizer channel, nozzle and nebulizer gas inlet fabricated on the silicon wafer, and a platinum heater sputtered on a glass wafer. These two wafers are joined by anodic bonding creating a two-dimensional version of an APCI microchip. The sample from GC is directed via heated transfer line capillary to the vaporizer channel of the APCI chip. The etched nozzle forms narrow sample plume, which is ionized by an external corona discharge needle, and the ions are analyzed by a mass spectrometer. The GC-microchip APCI-MS combination provides an efficient method for qualitative and quantitative analysis. The spectra produced by microchip APCI show intensive protonated molecule and some fragmentation products as in classical chemical ionization for structure elucidation. In quantitative analysis the GC-microchip APCI-MS showed good linearity (r(2) = 0.9989) and repeatability (relative standard deviation 4.4%). The limits of detection with signal-to-noise ratio of three were between 0.5 and 2 micromol/L with MS mode using selected ion monitoring and 0.05 micromol/L with MS/MS using multiple reaction monitoring.

  10. Liquid phase chromatography on microchips.

    Science.gov (United States)

    Kutter, Jörg P

    2012-01-20

    Over the past twenty years, the field of microfluidics has emerged providing one of the main enabling technologies to realize miniaturized chemical analysis systems, often referred to as micro-Total Analysis Systems (uTAS), or, more generally, Lab-on-a-Chip Systems (LOC) [1,2]. While microfluidics was driven forward a lot from the engineering side, especially with respect to ink jet and dispensing technology, the initial push and interest from the analytical chemistry community was through the desire to develop miniaturized sensors, detectors, and, very early on, separation systems. The initial almost explosive development of, in particular, chromatographic separation systems on microchips, has, however, slowed down in recent years. This review takes a closer, critical look at how liquid phase chromatography has been implemented in miniaturized formats over the past several years, what is important to keep in mind when developing or working with separations in a miniaturized format, and what challenges and pitfalls remain.

  11. Liquid phase chromatography on microchips

    DEFF Research Database (Denmark)

    Kutter, Jörg Peter

    2012-01-01

    Over the past twenty years, the field of microfluidics has emerged providing one of the main enabling technologies to realize miniaturized chemical analysis systems, often referred to as micro-Total Analysis Systems (uTAS), or, more generally, Lab-on-a-Chip Systems (LOC) [1,2]. While microfluidics...... was driven forward a lot from the engineering side, especially with respect to ink jet and dispensing technology, the initial push and interest from the analytical chemistry community was through the desire to develop miniaturized sensors, detectors, and, very early on, separation systems. The initial almost...... explosive development of, in particular, chromatographic separation systems on microchips, has, however, slowed down in recent years. This review takes a closer, critical look at how liquid phase chromatography has been implemented in miniaturized formats over the past several years, what is important...

  12. Detection of HCV core antigen and its diagnostic significance

    Directory of Open Access Journals (Sweden)

    YANG Jie

    2013-02-01

    Full Text Available ObjectiveTo compare the abilities of the hepatitis C virus (HCV core antigen (cAg test and the HCV RNA assay for confirming anti-HCV presence in order to determine the clinical utility of the HCV-cAg as an alternative or confirmatory diagnostic tool. MethodsSerum samples collected from 158 patients diagnosed with HCV infection were subjected to the enzyme-linked immunosorbent assay-based HCV-cAg test. The optical density (OD measured values were used to calculate the ratio of specimen absorbance to the cutoff value (S/CO. Simultaneously, the serum samples were subjected to PCR-based nucleic acid amplification quantitative fluorescence detection of HCV RNA. ResultsNone of the serum samples had a S/CO value <1 for the HCV-cAg test (100% negative, but all of the samples had a S/CO value >5 (100% positive. The HCV-cAg test sensitivity was 87.05%, specificity was 76.67%, positive predictive value was 9653%, and negative predictive value was 44.23%. As the S/CO value gradually increased, the significantly higher positive coincident rate of the HCV RNA test decreased. The HCV RNA negative coincident rate was significantly higher than that of the HCV-cAg test. HCV-cAg S/CO values between 1 and 2 corresponded to an HCV RNA values between 1.0×103 copies/ml and 1.0×104 copies/ml. The highest S/CO value obtained was 1.992. ConclusionThe HCV-cAg test is comparable to the HCV RNA assay for diagnosing HCV infection.

  13. Detection of Scabies: A Systematic Review of Diagnostic Methods

    Directory of Open Access Journals (Sweden)

    Victor Leung

    2011-01-01

    Full Text Available This study systematically reviewed the accuracy and precision of history, physical examination and tests for diagnosing scabies. Using a structured search strategy, Medline and Embase databases were searched for English and French language articles that included a diagnosis of scabies. Studies comparing history, physical examination and/or any diagnostic tests with the reference standard of microscopic visualization of mites, eggs or fecal elements obtained from skin scrapings or biopsies were included for analysis. History and examination of pruritic dermatoses failed to accurately diagnose scabies infection. The accuracy of dermatoscopy, performed by a trained practitioner, was determined; however, the accuracy of other diagnostic tests could not be calculated from the data in the literature. In the face of such diagnostic inaccuracy, research to identify or develop accurate diagnostic tests for scabies infection is needed and justifiable.

  14. Analysis of seven STR human loci for paternity testing by microchip electrophoresis

    Directory of Open Access Journals (Sweden)

    Karina Fraige

    2013-04-01

    Full Text Available The aim of this work was to evaluate two paternity cases by microchip electrophoresis and the validation of the methodology by comparison of the results with those obtained in a commercial genetic analyzer. It was observed that when working with tetranucleotide regions, in which the minimal difference between the alleles was only four base pairs, the commercial microchip system did not present the resolution and repeatability needed. Nevertheless, the relative standard deviation was between 0 and 1.2% and the fragments detected were within the expected size ranges as described in the literature.

  15. Challenges of glycoprotein analysis by microchip capillary gel electrophoresis.

    Science.gov (United States)

    Engel, Nicole; Weiss, Victor U; Wenz, Christian; Rüfer, Andreas; Kratzmeier, Martin; Glück, Susanne; Marchetti-Deschmann, Martina; Allmaier, Günter

    2015-08-01

    Glycosylations severely influence a protein's biological and physicochemical properties. Five exemplary proteins with varying glycan moieties were chosen to establish molecular weight (MW) determination (sizing), quantitation, and sensitivity of detection for microchip capillary gel electrophoresis (MCGE). Although sizing showed increasing deviations from literature values (SDS-PAGE or MALDI-MS) with a concomitant higher degree of analyte glycosylation, the reproducibility of MW determination and accuracy of quantitation with high sensitivity and reliability were demonstrated. Additionally, speed of analysis together with the low level of analyte consumption render MCGE attractive as an alternative to conventional SDS-PAGE. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Highly sensitive contactless conductivity microchips based on concentric electrodes for flow analysis.

    Science.gov (United States)

    Lima, Renato S; Piazzetta, Maria H O; Gobbi, Angelo L; Segato, Thiago P; Cabral, Murilo F; Machado, Sergio A S; Carrilho, Emanuel

    2013-12-18

    In this communication, we describe for the first time the integration of concentric electrodes (wrapping around the microchannel) in microchips. The use of such electrodes has been shown to be effective towards improvement of the sensitivity and detectability in pressure-driven flow platforms incorporating C(4)D.

  17. Rapid Isolation and Detection for RNA Biomarkers for TBI Diagnostics

    Science.gov (United States)

    2015-10-01

    CSR = transpose(chol(S)); % Cholesky Square Root of S CSR * transpose( CSR ) - S % Check for round-off errors A = inv( CSR ); % Mahalanobis scaling Z... transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers. Nature cell biology. 2008;10:1470-6. 57

  18. 78 FR 18988 - Establishing the Performance Characteristics of In Vitro Diagnostic Devices for the Detection of...

    Science.gov (United States)

    2013-03-28

    ... recommendations for studies to establish the analytical and clinical performance of in vitro diagnostic devices... studies for establishing the performance characteristics of in vitro diagnostic devices for the detection... HUMAN SERVICES Food and Drug Administration Establishing the Performance Characteristics of In...

  19. A mid-infrared absorption diagnostic for acetylene detection

    KAUST Repository

    KC, Utsav

    2015-05-14

    Acetylene is an important combustion intermediate and plays a critical role in soot formation. Accurate measurements of trace concentrations of acetylene can be very useful in validating hydrocarbon oxidation and soot formation mechanisms. Strongest vibrational band of acetylene near 13.7 μm is probed here to develop a highly sensitive absorption diagnostic. Experiments are carried out behind reflected shock waves to measure absorption cross sections of acetylene near 730 cm−1 over a wide range of temperatures (1000–2200 K) and pressures (1–5 bar). The diagnostic is demonstrated by measuring acetylene formation during the shock-heated pyrolysis and oxidation of propene. © 2015 Springer-Verlag Berlin Heidelberg

  20. Application of Microchip Electrophoresis for Clinical Tests

    Science.gov (United States)

    Yatsushiro, Shouki; Kataoka, Masatoshi

    Microchip electrophoresis has recently attracted much attention in the field of nuclear acid analysis due to its high efficiency, ease of operation, low consumption of samples and reagents, and relatively low costs. In addition, the analysis has expanded to an analytical field like not only the analysis of DNA but also the analysis of RNA, the protein, the sugar chain, and the cellular function, etc. In this report, we showed that high-performance monitoring systems for human blood glucose levels and α-amylase activity in human plasma using microchip electrophoresis.

  1. "Light-tagged" bacteriophage as a diagnostic tool for the detection of phytopathogens.

    Science.gov (United States)

    Schofield, David; Bull, Carolee T; Rubio, Isael; Wechter, W Patrick; Westwater, Caroline; Molineux, Ian J

    2013-01-01

    Detection of the phytopathogen Pseudomonas cannabina pv alisalensis, the causal agent of bacterial blight of crucifers is essential for managing this disease. A phage-based diagnostic assay was developed that detects and identifies P. cannabina pv alisalensis from cultures and diseased plant specimens. A recombinant "light-tagged" reporter phage was generated by integrating the luxAB genes into the P. cannabina pv alisalensis phage PBSPCA1 genome. PBSPCA1::luxAB is viable, stable and detects P. cannabina pv alisalensis within minutes and with high sensitivity by conferring a bioluminescent signal. Detection is dependent on cell viability since cells treated with a bactericidal disinfectant are unable to elicit a signal. Importantly, the reporter phage detects P. cannabina pv alisalensis from diseased plant specimens indicating the potential of the diagnostic for disease identification. The reporter phage displays promise for the rapid and specific diagnostic detection of cultivated isolates, and infected plant specimens.

  2. A capillary flow immunoassay microchip utilizing inkjet printing-based antibody immobilization onto island surfaces—toward sensitive and reproducible determination of carboxyterminal propeptide of type I procollagen

    Science.gov (United States)

    Fuchiwaki, Yusuke; Tanaka, Masato; Takaoka, Hiroki; Goya, Kenji

    2016-04-01

    A capillary-flow-driven microchip system requires no external power and has no moving off-chip components, in contrast with most microfluidic-based immunoassay systems which are complicated to operate and require external components. To accelerate the sensitive and reproducible determination of analytes required for practical point-of-care applications, we formed island microchannel surfaces on a microcapillary channel to allow stable antibody immobilization. The island surface was surrounded by a circular groove 10 μm deep and 150 μm wide and allowed uniform inkjet printing of antibody spots, complete bio-reagent replacement, and sensitive detection of luminescence intensity. Quantitative analysis of carboxyterminal propeptide of type I procollagen (PICP) concentrations using this microchannel was demonstrated between 0-600 ng·ml-1, which is adequate for the clinical estimation of PICP concentrations in the blood. This microchip system holds promise as a model diagnostic platform that is readily adaptable to hands-free operation.

  3. Host response to Clostridium difficile infection: Diagnostics and detection.

    Science.gov (United States)

    Usacheva, Elena A; Jin, Jian-P; Peterson, Lance R

    2016-12-01

    Clostridium difficile infection (CDI) is a significant healthcare concern worldwide, and C. difficile is recognised as the most frequent aetiological agent of infectious healthcare-associated diarrhoea in hospitalised adult patients. The clinical manifestation of CDI varies from self-limited diarrhoea to life-threatening colitis. Such a broad disease spectrum can be explained by the impact of host factors. Currently, a complex CDI aetiology is widely accepted, acknowledging the interaction between bacteria and the host. C. difficile strains producing clostridial toxins A and B are considered toxigenic and can cause disease; those not producing the toxins are non-pathogenic. A person colonised with a toxigenic strain will not necessarily develop CDI. It is imperative to recognise patients with active disease from those only colonised with this pathogen and to implement appropriate treatment. This can be achieved by diagnostics that rely on host factors specific to CDI. This review will focus on major aspects of CDI pathogenesis and molecular mechanisms, describing host factors in disease progression and assessment of the host response in order to facilitate the development of CDI-specific diagnostics.

  4. Scintillator diagnostics for the detection of laser accelerated ion beams

    Science.gov (United States)

    Cook, N.; Tresca, O.; Lefferts, R.

    2014-09-01

    Laser plasma interaction with ultraintense pulses present exciting schemes for accelerating ions. One of the advantages conferred by using a gaseous laser and target is the potential for a fast (several Hz) repetition rate. This requires diagnostics which are not only suited for a single shot configuration, but also for repeated use. We consider several scintillators as candidates for an imaging diagnostic for protons accelerated to MeV energies by a CO2 laser focused on a gas jet target. We have measured the response of chromium-doped alumina (chromox) and polyvinyl toluene (PVT) screens to protons in the 2-8 MeV range. We have calibrated the luminescent yield in terms of photons emitted per incident proton for each scintillator. We also discuss how light scattering and material properties affect detector resolution. Furthermore, we consider material damage and the presence of an afterglow under intense exposures. Our analysis reveals a near order of magnitude greater yield from chromox in response to proton beams at > 8 MeV energies, while scattering effects favor PVT-based scintillators at lower energies.

  5. Evaluation of microchip material and surface treatment options for IEF of allergenic milk proteins on microchips.

    Science.gov (United States)

    Poitevin, Martine; Shakalisava, Yuliya; Miserere, Sandrine; Peltre, Gabriel; Viovy, Jean-Louis; Descroix, Stephanie

    2009-12-01

    The use of glass and PDMS microchips has been investigated to perform rapid and efficient separation of allergenic whey proteins by IEF. To decrease EOF and to limit protein adsorption, two coating procedures have been compared. The first one consists in immobilizing hydroxypropyl cellulose (HPC) and the second one poly(dimethylacrylamide-co-allyl glycidyl ether) (PDMA-AGE). EOF limitation has been evaluated using frontal electrophoresis of a fluorescent marker of known effective mobility. EOF velocity was decreased by a factor about 100 and 30, respectively. pH gradient formation has been evaluated for each microchip using fluorescent pI markers. It was demonstrated that as expected a coating was essential to avoid pH gradient drift. Both coatings were efficient on glass microchips, but only PDMA-AGE allowed satisfying focusing of pI markers on PDMS microchips. Fluorescent covalent and noncovalent labelings of milk proteins have been compared by IEF on slab-gels. IEF separation of three major allergenic whey proteins [beta-lactoglobulin A (pI 5.25) and B (pI 5.35) and alpha-lactalbumin (pI 4.2-4.5)] was performed in both microchips. Milk proteins were separated with better resolution and shorter analysis time than by classical CIEF. Finally, better resolutions for milk allergens separation were obtained on glass microchips.

  6. Photopatterning of hydrogel microarryas in closed microchips

    NARCIS (Netherlands)

    Gümüscü, B.; Bomer, Johan G.; van den Berg, Albert; Eijkel, Jan C.T.

    2015-01-01

    To date, optical lithography has been extensively used for in situ patterning of hydrogel structures in a scale range from hundreds of microns to a few millimeters. The two main limitations which prevent smaller feature sizes of hydrogel structures are (1) the upper glass layer of a microchip

  7. Airy beams from a microchip laser

    CERN Document Server

    Longhi, Stefano

    2011-01-01

    It is theoretically shown that an end-pumped microchip laser formed by a thin laser crystal with plane-plane but slightly tilted facets can emit, under appropriate pumping conditions and near a crystal edge, a truncated self-accelerating Airy output beam.

  8. Photopatterning of hydrogel microarryas in closed microchips

    NARCIS (Netherlands)

    Gumuscu, Burcu; Bomer, Johan G.; Berg, van den Albert; Eijkel, Jan C.T.

    2015-01-01

    To date, optical lithography has been extensively used for in situ patterning of hydrogel structures in a scale range from hundreds of microns to a few millimeters. The two main limitations which prevent smaller feature sizes of hydrogel structures are (1) the upper glass layer of a microchip mainta

  9. Detection of novel diagnostic antibodies in ankylosing spondylitis: An overview.

    Science.gov (United States)

    Quaden, Dana H F; De Winter, Liesbeth M; Somers, Veerle

    2016-08-01

    Ankylosing spondylitis (AS) is a debilitating, chronic, rheumatic disease characterized by inflammation and new bone formation resulting in fusion of the spine and sacroiliac joints. Since early treatment is impeded by a delayed diagnosis, it is highly important to find new biomarkers that improve early diagnosis and may also contribute to a better assessment of disease activity, prognosis and therapy response in AS. Because of the absence of rheumatoid factor, AS was long assumed to have a seronegative character and antibodies are thus not considered a hallmark of the disease. However, emerging evidence suggests plasma cells and autoantibodies to be involved in the disease course. In this review, the role of B cells and antibodies in AS is discussed. Furthermore, an overview is provided of antibodies identified in AS up till now, and their diagnostic potential. Many of these antibody responses were based on small study populations and further validation is lacking. Moreover, most were identified by a hypothesis-driven approach and thus limited to antibodies against targets that are already known to be involved in AS pathogenesis. Hence, we propose an unbiased approach to identify novel diagnostic antibodies. The already successfully applied techniques cDNA phage display and serological antigen selection will be used to identify antibodies against both known and new antigen targets in AS plasma. These newly identified antibodies will enhance early diagnosis of AS and provide more insight into the underlying disease pathology, resulting in a more effective treatment strategy and eventually an improved disease outcome. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Engineering the bioelectrochemical interface using functional nanomaterials and microchip technique toward sensitive and portable electrochemical biosensors.

    Science.gov (United States)

    Jia, Xiaofang; Dong, Shaojun; Wang, Erkang

    2016-02-15

    Electrochemical biosensors have played active roles at the forefront of bioanalysis because they have the potential to achieve sensitive, specific and low-cost detection of biomolecules and many others. Engineering the electrochemical sensing interface with functional nanomaterials leads to novel electrochemical biosensors with improved performances in terms of sensitivity, selectivity, stability and simplicity. Functional nanomaterials possess good conductivity, catalytic activity, biocompatibility and high surface area. Coupled with bio-recognition elements, these features can amplify signal transduction and biorecognition events, resulting in highly sensitive biosensing. Additionally, microfluidic electrochemical biosensors have attracted considerable attention on account of their miniature, portable and low-cost systems as well as high fabrication throughput and ease of scaleup. For example, electrochemical enzymetic biosensors and aptamer biosensors (aptasensors) based on the integrated microchip can be used for portable point-of-care diagnostics and environmental monitoring. This review is a summary of our recent progress in the field of electrochemical biosensors, including aptasensors, cytosensors, enzymatic biosensors and self-powered biosensors based on biofuel cells. We presented the advantages that functional nanomaterials and microfluidic chip technology bring to the electrochemical biosensors, together with future prospects and possible challenges.

  11. Automated parallel DNA sequencing on multiple channel microchips.

    Science.gov (United States)

    Liu, S; Ren, H; Gao, Q; Roach, D J; Loder, R T; Armstrong, T M; Mao, Q; Blaga, I; Barker, D L; Jovanovich, S B

    2000-05-09

    We report automated DNA sequencing in 16-channel microchips. A microchip prefilled with sieving matrix is aligned on a heating plate affixed to a movable platform. Samples are loaded into sample reservoirs by using an eight-tip pipetting device, and the chip is docked with an array of electrodes in the focal plane of a four-color scanning detection system. Under computer control, high voltage is applied to the appropriate reservoirs in a programmed sequence that injects and separates the DNA samples. An integrated four-color confocal fluorescent detector automatically scans all 16 channels. The system routinely yields more than 450 bases in 15 min in all 16 channels. In the best case using an automated base-calling program, 543 bases have been called at an accuracy of >99%. Separations, including automated chip loading and sample injection, normally are completed in less than 18 min. The advantages of DNA sequencing on capillary electrophoresis chips include uniform signal intensity and tolerance of high DNA template concentration. To understand the fundamentals of these unique features we developed a theoretical treatment of cross-channel chip injection that we call the differential concentration effect. We present experimental evidence consistent with the predictions of the theory.

  12. Quantitative analysis of plasma interleiukin-6 by immunoassay on microchip

    Science.gov (United States)

    Abe, K.; Hashimoto, Y.; Yatsushiro, S.; Yamamura, S.; Tanaka, M.; Ooie, T.; Baba, Y.; Kataoka, M.

    2012-03-01

    Sandwich enzyme-linked immunoassay (ELISA) is one of the most frequently employed assays for clinical diagnosis, since this enables the investigator to identify specific protein biomarkers. However, the conventional assay using a 96-well microtitration plate is time- and sample-consuming, and therefore is not suitable for rapid diagnosis. To overcome these drawbacks, we performed a sandwich ELISA on a microchip. We employed the piezoelectric inkjet printing for deposition and fixation of 1st antibody on the microchannnel surface (300 μm width and 100 μm depth). Model analyte was interleukin-6 (IL-6) which was one of the inflammatory cytokine. After blocking the microchannel, antigen, biotin-labeled 2nd antibody, and avidin-labeled peroxidase were infused into the microchannel and incubated for 20 min, 10 min, and 5 min, respectively. This assay could detect 2 pg/ml and quantitatively measure the range of 0-32 pg/ml. Liner regression analysis of plasma IL-6 concentration obtained by microchip and conventional methods exhibited a significant relationship (R2 = 0.9964). This assay reduced the time for the antigen-antibody reaction to 1/6, and the consumption of samples and reagents to 1/50 compared with the conventional method. This assay enables us to determine plasma IL-6 with accuracy, high sensitivity, time saving ability, and low consumption of sample and reagents, and thus will be applicable to clinic diagnosis.

  13. Diagnostic aids for detection of oral precancerous conditions

    Institute of Scientific and Technical Information of China (English)

    Diana V Messadi

    2013-01-01

    Oral cancer has a tendency to be detected at late stage which is detrimental to the patients because of its high mortality and morbidity rates. Early detection of oral cancer is therefore important to reduce the burden of this devastating disease. In this review article, the most common oral precancerous lesions are discussed and the importance of early diagnosis is emphasized. In addition, the most common non-invasive oral cancer devices that can aid the general practitioners in early diagnosis are also discussed.

  14. Convenient diagnosis of spinal and bulbar muscular atrophy using a microchip electrophoresis system.

    Science.gov (United States)

    Maruyama, Hirofumi; Morino, Hiroyuki; Izumi, Yuishin; Noda, Kouichi; Kawakami, Hideshi

    2013-01-01

    Spinal and bulbar muscular atrophy (SBMA) is a slowly progressive motor neuron disease. Lower and primary sensory neuronopathy is one of the major neuropathological changes that occurs in SBMA. However, many sings are common to SBMA and amyotrophic lateral sclerosis (ALS), and SBMA patients are sometimes diagnosed with ALS. Leuprorelin may be used to treat SBMA, but an accurate diagnosis is necessary for treatment and care. Genetic diagnosis can be performed to detect the expansion of a CAG repeat in the androgen receptor gene in SBMA patients. To screen for this expansion, we used a microchip electrophoresis system. The discrepancy between the actual repeat length and that found by the microchip electrophoresis system was roughly dependent on the repeat length. The mean difference was -6.8 base pairs (bp) in SBMA patients, -0.30 bp in controls. The microchip electrophoresis results were approximately 2 CAG repeats shorter than the actual repeat length in SBMA patients. Using this method, we screened our ALS samples (31 were familial, 271 were sporadic): 4 subjects were diagnosed with SBMA; 2 had familial ALS, and 2 had sporadic ALS (0.7%). The microchip electrophoresis system is semi-quantitative, convenient and useful for screening a large number of samples.

  15. 76 FR 69034 - Microbiology Devices; Classification of In Vitro Diagnostic Device for Yersinia Species Detection

    Science.gov (United States)

    2011-11-07

    ... devices with similar risk profiles. Reagents for detection of specific novel influenza A viruses, for... of Health and Human Services Food and Drug Administration 21 CFR Part 866 Microbiology Devices...] Microbiology Devices; Classification of In Vitro Diagnostic Device for Yersinia Species Detection AGENCY: Food...

  16. Interferometric Reflectance Imaging Sensor (IRIS—A Platform Technology for Multiplexed Diagnostics and Digital Detection

    Directory of Open Access Journals (Sweden)

    Oguzhan Avci

    2015-07-01

    Full Text Available Over the last decade, the growing need in disease diagnostics has stimulated rapid development of new technologies with unprecedented capabilities. Recent emerging infectious diseases and epidemics have revealed the shortcomings of existing diagnostics tools, and the necessity for further improvements. Optical biosensors can lay the foundations for future generation diagnostics by providing means to detect biomarkers in a highly sensitive, specific, quantitative and multiplexed fashion. Here, we review an optical sensing technology, Interferometric Reflectance Imaging Sensor (IRIS, and the relevant features of this multifunctional platform for quantitative, label-free and dynamic detection. We discuss two distinct modalities for IRIS: (i low-magnification (ensemble biomolecular mass measurements and (ii high-magnification (digital detection of individual nanoparticles along with their applications, including label-free detection of multiplexed protein chips, measurement of single nucleotide polymorphism, quantification of transcription factor DNA binding, and high sensitivity digital sensing and characterization of nanoparticles and viruses.

  17. Prenetal Detection of Oral Clefts : Diagnostic, Genetic and Ethical Aspects

    NARCIS (Netherlands)

    Maarse, W.

    2015-01-01

    Since the introduction of routine prenatal screening with ultrasound in the Netherlands in 2007, parents are confronted with the diagnosis of oral cleft (OC) already during pregnancy. This imposed a new dimension in cleft care in the Netherlands. As a consequence to increasing prenatal detection rat

  18. Diagnostic Methods for Detection of Blood-Borne Candidiasis.

    Science.gov (United States)

    Clancy, Cornelius J; Nguyen, M Hong

    2016-01-01

    β-D-glucan (Fungitell) and polymerase chain reaction-based (T2Candida) assays of blood samples are FDA-approved adjuncts to cultures for diagnosing candidemia and other types of invasive candidiasis, but their clinical roles are unclear. In this chapter, we describe laboratory protocols for performing Fungitell and T2Candida assays. We then discuss step-by-step methods for interpreting test results at the bedside using a Bayesian framework, and for incorporating assays into rational patient management strategies. Prior to interpreting results, clinicians must recognize that test performance varies based on the type of invasive candidiasis being diagnosed. In general, the type of invasive candidiasis that is most likely in a given patient can be identified, and the pretest likelihood of disease estimated. From there, positive and negative predictive values (PPV, NPV) for an assay can be calculated. At a population level, tests can be incorporated into screening strategies for antifungal treatment. NPV and PPV thresholds can be defined for discontinuing antifungal prophylaxis or initiating preemptive treatment, respectively. Using the thresholds, it is possible to assign windows of pretest likelihood for invasive candidiasis (and corresponding patient populations) in which tests are most likely to valuable. At the individual patient level, tests may be useful outside of the windows proposed for screening populations. The interpretive and clinical decision-making processes we discuss will be applicable to other diagnostic assays as they enter the clinic, and to existing assays as more data emerge from various populations.

  19. Detection of mycobacterial siderophores and implications for diagnostics

    OpenAIRE

    McBride, Nicholas S.

    2012-01-01

    Mycobacterium tuberculosis (Mtb) is a pervasive human pathogen that continues to kill 1.8 million people every year. Acquisition of iron within the host is vital to the pathogenicity of Mtb and to accomplish this it produces siderophores called mycobactins and carboxymycobactins. This dissertation has sought to build a simple assay for detection and quantification of mycobacterial siderophores that has potential applications in tuberculosis (TB) diagnosis. This assay has been tested a...

  20. Diagnostic Inaccuracy of Smart Phone Applications for Melanoma Detection

    Science.gov (United States)

    Wolf, Joel; Moreau, Jacqui; Akilov, Oleg; Patton, Timothy; English, Joseph C; Ho, Jon; Ferris, Laura Korb

    2013-01-01

    Objective To measure the performance of smart phone applications which evaluate photographs of skin lesions and provide the user feedback as to their likelihood of malignancy. Design Case-control diagnostic accuracy study Setting Academic dermatology department Participants Digital clinical images of pigmented cutaneous lesions (60 melanoma cases and 128 benign lesion controls), all with histologic diagnosis rendered by a board-certified dermatopathologist, obtained prior to biopsy in patients undergoing lesion removal as part of routine care. Main Outcome Measures Sensitivity, specificity, and positive and negative predictive values of four smart phone applications designed to aid non-clinician users in determining if their skin lesion is benign or malignant. Results Sensitivity of the four tested applications ranged from 6.8% to 98.1%. Specificity ranged from 30.4% to 93.7%. Positive predictive value ranged from 33.3% to 42.1%, and negative predictive value ranged from 65.4% to 97.0%. The highest sensitivity for melanoma diagnosis was observed for an application that sends the image directly to a board-certified dermatologist for analysis and the lowest sensitivity was observed for applications that use automated algorithms to analyze images. Conclusions The performance of smart phone applications in assessing melanoma risk is highly variable, and 3 out of 4 smart phone applications incorrectly classified 30% or more of melanomas as unconcerning. Reliance on these applications, which are not subject to regulatory oversight, in lieu of medical consultation, has the potential to delay the diagnosis of melanoma and to harm users. PMID:23325302

  1. Comparison of surface and hydrogel-based protein microchips.

    Science.gov (United States)

    Zubtsov, D A; Savvateeva, E N; Rubina, A Yu; Pan'kov, S V; Konovalova, E V; Moiseeva, O V; Chechetkin, V R; Zasedatelev, A S

    2007-09-15

    Protein microchips are designed for high-throughput evaluation of the concentrations and activities of various proteins. The rapid advance in microchip technology and a wide variety of existing techniques pose the problem of unified approach to the assessment and comparison of different platforms. Here we compare the characteristics of protein microchips developed for quantitative immunoassay with those of antibodies immobilized on glass surfaces and in hemispherical gel pads. Spotting concentrations of antibodies used for manufacturing of microchips of both types and concentrations of antigen in analyte solution were identical. We compared the efficiency of antibody immobilization, the intensity of fluorescence signals for both direct and sandwich-type immunoassays, and the reaction-diffusion kinetics of the formation of antibody-antigen complexes for surface and gel-based microchips. Our results demonstrate higher capacity and sensitivity for the hydrogel-based protein microchips, while fluorescence saturation kinetics for the two types of microarrays was comparable.

  2. Microchip capillary electrophoresis based electroanalysis of triazine herbicides.

    Science.gov (United States)

    Islam, Kamrul; Chand, Rohit; Han, Dawoon; Kim, Yong-Sang

    2015-01-01

    The number of pesticides used in agriculture is increasing steadily, leading to contamination of soil and drinking water. Herein, we present a microfluidic platform to detect the extent of contamination in soil samples. A microchip capillary electrophoresis system with in-channel electrodes was fabricated for label-free electroanalytical detection of triazine herbicides. The sample mixture contained three representative triazines: simazine, atrazine and ametryn. The electropherogram for each individual injection of simazine, atrazine and ametryn showed peaks at 58, 66 and 72 s whereas a mixture of them showed distinct peaks at 59, 67 and 71 s respectively. The technique as such may prove to be a useful qualitative and quantitative tool for the similar environmental pollutants.

  3. Diagnostic Accuracy of Ultrasonography and Radiography in Detection of Pulmonary Contusion; a Systematic Review and Meta-Analysis

    OpenAIRE

    HOSSEINI, Mostafa; Ghelichkhani, Parisa; Baikpour, Masoud; Tafakhori, Abbas; Asady, Hadi; Haji Ghanbari, Mohammad Javad; Yousefifard, Mahmoud; Safari, Saeed

    2015-01-01

    Introduction: Ultrasonography is currently being used as one of the diagnostic modalities in various medical emergencies for screening of trauma patients. The diagnostic value of this modality in detection of traumatic chest injuries has been evaluated by several studies but its diagnostic accuracy in diagnosis of pulmonary contusion is a matter of discussion. Therefore, the present study aimed to determine the diagnostic accuracy of ultrasonography and radiography in detection of pulmonary c...

  4. Microchip capillary electrophoresis-electrospray ionization-mass spectrometry of intact proteins using uncoated Ormocomp microchips.

    Science.gov (United States)

    Sikanen, Tiina; Aura, Susanna; Franssila, Sami; Kotiaho, Tapio; Kostiainen, Risto

    2012-01-20

    We present rapid (microchips. The microchips are fabricated fully of commercial inorganic-organic hybrid material, Ormocomp, by UV-embossing and adhesive Ormocomp-Ormocomp bonding (CE microchannels). A sheath-flow ESI interface is monolithically integrated with the UV-embossed separation channels by cutting a rectangular emitter tip in the end with a dicing saw. As a result, electrospray was produced from the corner of chip with good reproducibility between parallel tips (stability within 3.8-9.2% RSD). Thanks to its inherent biocompatibility and stable (negative) surface charge, Ormocomp microchips enable efficient intact protein analysis with up to ∼10(4) theoretical separation plates per meter without any chemical or physical surface modification before analysis. The same microchip setup is also feasible for rapid peptide sequencing and mass fingerprinting and shows excellent migration time repeatability from run to run for both peptides (5.6-5.9% RSD, n=4) and intact proteins (1.3-7.5% RSD, n=3). Thus, the Ormocomp microchips provide a versatile new tool for MS-based proteomics. Particularly, the feasibility of the Ormocomp chips for rapid analysis of intact proteins with such a simple setup is a valuable increment to the current technology.

  5. Evaluation of pain and inflammation associated with hot iron branding and microchip transponder injection in horses.

    Science.gov (United States)

    Lindegaard, Casper; Vaabengaard, Dorte; Christophersen, Mogens T; Ekstøm, Claus T; Fjeldborg, Julie

    2009-07-01

    To compare effects of hot iron branding and microchip transponder injection regarding aversive behavioral reactions indicative of pain and inflammation in horses. 7 adult horses. In a randomized controlled clinical crossover study, behavioral reactions to hot iron branding and microchip transponder injection were scored by 4 observers. Local and systemic inflammation including allodynia were assessed and compared by use of physiologic and biochemical responses obtained repeatedly for the 168-hour study period. Serum cortisol concentration was measured repeatedly throughout the first 24 hours of the study. Sham treatments were performed 1 day before and 7 days after treatments. Hot iron branding elicited a significantly stronger aversive reaction indicative of pain than did microchip transponder injection (odds ratio [OR], 12.83). Allodynia quantified by means of skin sensitivity to von Frey monofilaments was significantly greater after hot iron branding than after microchip transponder injection (OR, 2.59). Neither treatment induced signs of spontaneously occurring pain that were observed during the remaining study period, and neither treatment induced increased serum cortisol concentrations. Comparison with sham treatments indicated no memory of an unpleasant event. The hot iron branding areas had significantly increased skin temperature and swelling (OR, 14.6). Systemic inflammation as measured via serum amyloid A concentration was not detected after any of the treatments. Microchip transponder injection induced less signs of pain and inflammation and did not seem to pose a higher long-term risk than hot iron branding. Consequently, results indicated that hot iron branding does inflict more pain and should be abandoned where possible.

  6. New Computer Assisted Diagnostic to Detect Alzheimer Disease

    Directory of Open Access Journals (Sweden)

    Ben Rabeh Amira

    2016-08-01

    Full Text Available We describe a new Computer Assisted Diagnosis (CAD to automatically detect Alzheimer Patients (AD, Mild Cognitive Impairment (MCI and elderly Controls, based on the segmentation and classification of the Hippocampus (H and Corpus Calosum (CC from Magnetic Resonance Images (MRI. For the segmentation we used a new method based on a deformable model to extract the area wishes, and then we computed the geometric and texture features. For the classification we proposed a new supervised method. We evaluated the accuracy of our method in a group of 25 patients with AD (age±standard-deviation (SD =70±6 years, 25 patients with MCI (age±SD=65±8 years and 25 elderly healthy controls (age±SD=60±8 years. For the AD patients we found an accuracy of the classification of 92%, for the MCI we found 88% and for the elderly patients we found 96%. Overall, we found our method to be 92% accurate. Our method can be a useful tool for diagnosing Alzheimer’s Disease in any of these Steps.

  7. Towards a portable microchip system with integrated thermal control and polymer waveguides for real-time PCR

    DEFF Research Database (Denmark)

    Wang, Zhenyu; Sekulovic, Andrea; Kutter, Jörg Peter

    2006-01-01

    performed with a dilution series of C. jejuni DNA template (2 to 200 pg/mu l) could be quantitatively detected and compared with a conventional post-PCR analysis (DNA gel electrophoresis). The presented approach provided reliable real-time quantitative information of the PCR amplification of the targeted......A novel real-time PCR microchip platform with integrated thermal system and polymer waveguides has been developed. The integrated polymer optical system for real-time monitoring of PCR was fabricated in the same SU-8 layer as the PCR chamber, without additional masking steps. Two suitable DNA...... binding dyes, SYTOX Orange and TO-PRO-3, were selected and tested for the real-time PCR processes. As a model, cadF gene of Campylobacter jejuni has been amplified on the microchip. Using the integrated optical system of the real-time PCR microchip, the measured cycle threshold values of the real-time PCR...

  8. Quantitative polymerase chain reaction (PCR) for detection of aquatic animal pathogens in a diagnostic laboratory setting

    Science.gov (United States)

    Purcell, Maureen K.; Getchell, Rodman G.; McClure, Carol A.; Weber, S.E.; Garver, Kyle A.

    2011-01-01

    Real-time, or quantitative, polymerase chain reaction (qPCR) is quickly supplanting other molecular methods for detecting the nucleic acids of human and other animal pathogens owing to the speed and robustness of the technology. As the aquatic animal health community moves toward implementing national diagnostic testing schemes, it will need to evaluate how qPCR technology should be employed. This review outlines the basic principles of qPCR technology, considerations for assay development, standards and controls, assay performance, diagnostic validation, implementation in the diagnostic laboratory, and quality assurance and control measures. These factors are fundamental for ensuring the validity of qPCR assay results obtained in the diagnostic laboratory setting.

  9. The Diagnostic Value of Superb Microvascular Imaging (SMI) in Detecting Blood Flow Signals of Breast Lesions

    Science.gov (United States)

    Ma, Yan; Li, Gang; Li, Jing; Ren, Wei-dong

    2015-01-01

    Abstract The correlation between color Doppler flow imaging (CDFI) and Superb Microvascular Imaging (SMI) for detecting blood flow in breast lesions was investigated, as was the diagnostic value of SMI in differentiating benign from malignant breast lesions. These lesions were evaluated using both CDFI and SMI according to Adler's method. Pathologic examination showed 57 malignant lesions and 66 benign lesions. The number of blood vessels in a single mass was detected by 2 techniques (SMI and CDFI), and the difference between the 2 values (SMI-CDFI) was calculated. The optimal threshold for the diagnosis of malignant neoplasms and the diagnostic performances of SMI, CDFI, and SMI-CDFI were calculated. For the total lesions and malignant lesions alone, the difference between SMI and CDFI for detecting blood flow was significant (P < 0.01), but the difference was not significant for benign lesions (P = 0.15). The area under the receiver operating characteristic curve was 0.73 (95% confidence interval [CI]: 0.64–0.82) for CDFI; 0.81 (95% CI: 0.74–0.89) for SMI; and 0.89 (95% CI: 0.82–0.95) for SMI-CDFI. Furthermore, the modality of “SMI-CDFI” showed the best diagnostic performance. SMI provides further microvessel information in breast lesions. The diagnostic modality of “SMI-CDFI” can improve the diagnostic performance of ultrasound in the differentiation between benign and malignant masses. PMID:26356718

  10. Evaluation of a vibration diagnostic system for the detection of spur gear pitting failures

    Science.gov (United States)

    Townsend, Dennis P.; Zakrajsek, James J.

    1993-06-01

    A vibration diagnostic system was used to detect spur gear surface pitting fatigue in a closed-loop spur gear fatigue test rig. The diagnostic system, comprising a personal computer with an analog-to-digital conversion board, a diagnostic system unit, and software, uses time-synchronous averaging of the vibration signal to produce a vibration image of each tooth on any gear in a transmission. Several parameters were analyzed including gear pair stress wave and raw baseband vibration, kurtosis, peak ratios, and others. The system provides limits for the various parameters and gives a warning when the limits are exceeded. Several spur gear tests were conducted with this system and vibration data analyzed at 5-min. intervals. The results presented herein show that the system is fairly effective at detecting spur gear tooth surface fatigue pitting failures.

  11. Comparative Capabilities of Clinical Assessment, Diagnostic Criteria, and Polysomnography in Detecting Sleep Bruxism

    Science.gov (United States)

    Palinkas, Marcelo; De Luca Canto, Graziela; Rodrigues, Laíse Angélica Mendes; Bataglion, César; Siéssere, Selma; Semprini, Marisa; Regalo, Simone Cecilio Hallak

    2015-01-01

    Objective: To evaluate the diagnostic capability of signs and symptoms of sleep bruxism (SB) as per the American Academy of Sleep Medicine (AASM) criteria and a diagnostic grading system proposed by international experts for assessing SB. Methods: The study was conducted in three phases (interview, physical examination, and sleep studies). Subjects were asked about self-reported tooth grinding sounds occurring during sleep, muscle fatigue, temporal headaches, jaw muscle pain, and jaw locking. A visual examination was conducted to check for presence of abnormal tooth wear. A full-night polysomnography (PSG) was performed. After three phases, the subjects were divided into two groups matched by age and gender: Case Group, 45 SB subjects, and Control Group, 45 non-SB subjects. Diagnostic accuracy measurements were calculated for each sign or symptom individually and for the two diagnostic criteria analyzed. Results: Muscle fatigue, temporal headaches, and AASM criteria were associated with highest sensitivity (78%, 67%, 58%, respectively) and also with highest diagnostic odds ratio (OR = 9.63, 9.25, 6.33, respectively). Jaw locking, muscle pain, and the criterion of “probable SB” were associated with the worst sensitivity (16%, 18%, 22%, respectively). Conclusions: Presence of muscle fatigue and temporal headaches can be considered good tools to screen SB patients. None of the diagnostic criteria evaluated was able to accurately identify patients with SB. AASM criteria had the strongest diagnostic capabilities and—although they do not attain diagnostic values high enough to replace the current gold standard (PSG)—should be used as a screening tool to identify SB. Citation: Palinkas M, De Luca Canto G, Rodrigues LA, Bataglion C, Siéssere S, Semprini M, Regalo SC. Comparative capabilities of clinical assessment, diagnostic criteria, and polysomnography in detecting sleep bruxism. J Clin Sleep Med 2015;11(11):1319–1325. PMID:26235152

  12. Implantable microchip transponders for body temperature measurements in pigs

    DEFF Research Database (Denmark)

    Lohse, Louise; Uttenthal, Åse; Enøe, Claes

    Objective Body temperature is a simple, but clinically important parameter in monitoring the health status of pigs, both at individual level and herd level. The standard procedure for obtaining such data is normally performed by recording of the core body temperature, using a rectal digital...... with CSFV, as determined by virus detection. So, the transponder system was tested in both clinically healthy and clinically ill pigs with physiologically normal body temperature or fever, respectively. The data obtained in this study, showed a correlation between the two methods for monitoring body...... body temperature was tested, in order to evaluate the utility and reliability of this tool, in domestic pigs. The system is presently used and well optimized in small laboratory animals [1, 2]. We tested the microchip transponders during experimental infection of pigs with classical swine fever virus...

  13. A novel technique for detecting antibiotic-resistant typhoid from rapid diagnostic tests.

    OpenAIRE

    2015-01-01

    Fluoroquinolone-resistant typhoid is increasing. An antigen-detecting rapid diagnostic test (RDT) can rapidly diagnose typhoid from blood cultures. A simple, inexpensive molecular technique performed with DNA from positive RDTs accurately identified gyrA mutations consistent with phenotypic susceptibility testing results. Field diagnosis combined with centralized molecular resistance testing could improve typhoid management and surveillance in low-resource settings.

  14. Surprise, p-value, s-value and a diagnostic procedure to detect not informative experiments

    Science.gov (United States)

    Recchia, Daniela R.; Ostermann, Thomas; Garcia, Jesus E.

    2016-06-01

    In this paper, examples of diverse measures of significance are studied in the frame of Neyman-Pearson hypothesis tests. A diagnostic procedure is proposed, to detect non informative experiments where the p-value procedure might fail to measure the significance of the outcome of an experiment.

  15. Laboratory Evaluation of Three Rapid Diagnostic Tests for Dual Detection of HIV and Treponema pallidum Antibodies

    OpenAIRE

    Humphries, Romney M.; Woo, Jennifer S.; Chung, Jun Ho; Sokovic, Anita; Bristow, Claire C; Jeffrey D Klausner

    2014-01-01

    The performance of three research-use-only, dual HIV and syphilis rapid diagnostic tests (RDTs) was evaluated for 150 patient serum samples and compared to reference HIV and Treponema pallidum antibody detection methods. The RDTs performed comparably, with sensitivities of 93 to 99% and specificities of 97 to 100%. The kappa statistic between the RDTs was 0.95.

  16. Detection of distant metastases in patients with oesophageal or gastric cardia cancer: A diagnostic decision analysis

    NARCIS (Netherlands)

    E.P.M. van Vliet (Evelyn); E.W. Steyerberg (Ewout); M.J.C. Eijkemans (René); E.J. Kuipers (Ernst); P.D. Siersema (Peter)

    2007-01-01

    textabstractComputed tomography (CT) is presently a standard procedure for the detection of distant metastases in patients with oesophageal or gastric cardia cancer. We aimed to determine the additional diagnostic value of alternative staging investigations. We included 569 oesophageal or gastric

  17. Diagnostic value of respiratory virus detection in symptomatic children using real-time PCR

    NARCIS (Netherlands)

    Huijskens, Elisabeth G; Biesmans, Renée C; Buiting, Anton G; Obihara, Charles C; Rossen, John W

    2012-01-01

    Background: Acute respiratory tract infections are an important public health problem. Sensitive and rapid diagnostic techniques have been developed and are used in daily clinical practice. Here we evaluate the clinical relevance of detecting 20 common respiratory pathogens by molecular methods in a

  18. Diagnostic accuracy of whole-brain CT perfusion in the detection of acute infratentorial infarctions

    Energy Technology Data Exchange (ETDEWEB)

    Bollwein, Christine; Sommer, Wieland H.; Thierfelder, Kolja M.; Reiser, Maximilian F. [Ludwig-Maximilians-University Hospital of Munich, Institute for Clinical Radiology, Munich (Germany); Plate, Annika; Straube, Andreas; Baumgarten, Louisa von [Ludwig-Maximilians-University Hospital of Munich, Department of Neurology, Munich (Germany); Janssen, Hendrik [South Nuremberg Hospital, Department of Neuroradiology, Nuremberg (Germany)

    2016-11-15

    Although the diagnostic performance of whole-brain computed tomographic perfusion (WB-CTP) in the detection of supratentorial infarctions is well established, its value in the detection of infratentorial strokes remains less well defined. We examined its diagnostic accuracy in the detection of infratentorial infarctions and compared it to nonenhanced computed tomography (NECT), aiming to identify factors influencing its detection rate. Out of a cohort of 1380 patients who underwent WB-CTP due to suspected stroke, we retrospectively included all patients with MRI-confirmed infratentorial strokes and compared it to control patients without infratentorial strokes. Two blinded readers evaluated NECT and four different CTP maps independently for the presence and location of infratentorial ischemic perfusion deficits. The study was designed as a retrospective case-control study and included 280 patients (cases/controls = 1/3). WB-CTP revealed a greater diagnostic sensitivity than NECT (41.4 vs. 17.1 %, P = 0.003). The specificity, however, was comparable (93.3 vs. 95.0 %). Mean transit time (MTT) and time to drain (TTD) were the most sensitive (41.4 and 40.0 %) and cerebral blood volume (CBV) the most specific (99.5 %) perfusion maps. Infarctions detected using WB-CTP were significantly larger than those not detected (15.0 vs. 2.2 ml; P = 0.0007); infarct location, however, did not influence the detection rate. The detection of infratentorial infarctions can be improved by assessing WB-CTP as part of the multimodal stroke workup. However, it remains a diagnostic challenge, especially small volume infarctions in the brainstem are likely to be missed. (orig.)

  19. Encapsulated electrodes for microchip devices: microarrays and platinized electrodes for signal enhancement.

    Science.gov (United States)

    Selimovic, Asmira; Martin, R Scott

    2013-07-01

    In this paper, we present two new methodologies of improving the performance of microchip-based electrochemical detection in microfluidic devices. The first part describes the fabrication and characterization of epoxy-embedded gold microelectrode arrays that are evenly spaced and easily modified. Electrodepositions using a gold plating solution can be performed on the electrodes to result in a 3D pillar array that, when used with microchip-based flow injection analysis, leads to an eightfold increase in signal (when compared to a single electrode), with the LOD for catechol being 4 nM. For detecting analytically challenging molecules such as nitric oxide (NO), platinization of electrodes is commonly used to increase the sensitivity. It is shown here that microchip devices containing either the pillar arrays or more traditional glassy carbon electrodes can be modified with platinum black (Pt-black) for NO detection. In the case of using glassy carbon electrodes for NO detection, integration of the resulting platinized electrode with microchip-based flow analysis resulted in a ten times signal increase relative to use of a bare glassy carbon electrode. In addition, it is demonstrated that these electrodes can be coated with Nafion to impart selectivity toward NO over interfering species such as nitrite. The LOD for NO when using the Pt-black /Nafion-coated glassy carbon electrode was 9 nM. These electrodes can also be embedded in a polystyrene substrate, with the applicability of these sensitive and selective electrodes being demonstrated by monitoring the adenosine triphosphate-mediated release of NO from endothelial cells immobilized in a microfluidic network without any adhesion factor.

  20. Assessment of Urinary-5-Hydroxyindolacetic Acid as A Diagnostic Parameter in Early Detection of Acute Appendicitis

    Directory of Open Access Journals (Sweden)

    Zuhair B Kamal

    2017-04-01

    Full Text Available Background: Acute appendicitis is the most common abdominal surgical emergency especially in children and young adults. The diagnosis of appendicitis is difficult because half the cases are incorrectly identified. Serotonin was defined as a good diagnostic marker for many inflammations including appendicitis and it is metabolite into 5-hydroxyindolacetic acid (5-HIAA to be excreted in urine. 5-HIAA is suggested to be of diagnostic importance in the detection of this disease. The aim of this study was to evaluate the diagnostic importance of urinary-5-HIAA as an added parameter to Alvarado score. Methods: Seventy patients (35 females and 35 males with acute appendicitis (35 were mild and 35 severe-perforated and gangrenous were included in this study and 70 healthy individuals were taken as a control group. Urinary-5-HIAA was estimated in all patients and control group using ELISA method. Results: Sensitivity for the mild group is 94.2%, specificity 100% and diagnostic accuracy is 97.4%, while the sensitivity for the severe group is 37%. It was found that there is a highly significant difference between mild and control groups (P<0.05. The diagnostic accuracy for the mild group is 97.4% and for the severe is 68.5%. Conclusion: We conclude that urinary-5-HIAA is a high sensitive test for early detection of acute appendicitis.

  1. In-situ photopatterning of hydrogel microarrays in polished microchips

    NARCIS (Netherlands)

    Gümüscü, B.; van den Berg, Albert; Eijkel, Jan C.T.

    2015-01-01

    We present a fabrication method which enables simple and eproducible photopatterning of micron- sized hydrogel arrays inside closed microchips. To achieve this, the glass cover of the microchip is thinned by mechanical grinding and polishing. This procedure reduces the spacing between the photomask

  2. Adding functionality to microchips by wafer post-processing

    NARCIS (Netherlands)

    Schmitz, J.

    2007-01-01

    The traditional microchip processes, stores and communicates electrical information. Here we review an emerging class of microchips that have additional functionality through extra integrated components in the chip. In the final manufacturing stage, layers are added on top of the chip, with a specif

  3. In-situ photopatterning of hydrogel microarrays in polished microchips

    NARCIS (Netherlands)

    Gumuscu, Burcu; Berg, van den Albert; Eijkel, Jan C.T.

    2015-01-01

    We present a fabrication method which enables simple and eproducible photopatterning of micron- sized hydrogel arrays inside closed microchips. To achieve this, the glass cover of the microchip is thinned by mechanical grinding and polishing. This procedure reduces the spacing between the photomask

  4. Adding functionality to microchips by wafer post-processing

    NARCIS (Netherlands)

    Schmitz, Jurriaan

    2007-01-01

    The traditional microchip processes, stores and communicates electrical information. Here we review an emerging class of microchips that have additional functionality through extra integrated components in the chip. In the final manufacturing stage, layers are added on top of the chip, with a specif

  5. Comparison between Amnisure Placental Alpha Microglobulin-1 Rapid Immunoassay and Standard Diagnostic Methods for Detection of Rupture of Membranes

    OpenAIRE

    Beng Kwang Ng; Pei Shan Lim; Mohamad Nasir Shafiee; Nur Azurah Abdul Ghani; Nor Azlin Mohamed Ismail; Mohd Hashim Omar; Muhammad Abdul Jamil Muhammad Yassin

    2013-01-01

    Objective. To determine the diagnostic accuracy of placental alpha microglobulin-1 assay and standard diagnostic methods for detecting rupture of membrane. Study Design. Prospective diagnostic study, between June 2011 to November 2011 at a tertiary centre. Initial evaluation included both the standard diagnostic methods for rupture of membranes and placental alpha microglobulin-1 immunoassay. The actual rupture of membranes was diagnosed on review of the medical records after delivery (absenc...

  6. Bulk modification of PDMS microchips by an amphiphilic copolymer.

    Science.gov (United States)

    Xiao, Yan; Yu, Xiao-Dong; Xu, Jing-Juan; Chen, Hong-Yuan

    2007-09-01

    A simple and rapid bulk-modification method based on adding an amphiphilic copolymer during the fabrication process was employed to modify PDMS microchips. Poly(lactic acid)-poly(ethylene glycol) (PLA-PEG) was used as the additive substance. Compared to the native PDMS microchips, both the contact angle and the EOF of the bulk-modified PDMS microchips decreased. The effects of the additive loading and the pH on the EOF were investigated in detail. The bulk-modified PDMS microchips exhibited reproducible and stable EOF behavior. The application of the bulk-modified PDMS microchips was also studied and the results indicated that they could be successfully used to separate amino acids and to suppress protein adsorption.

  7. Diagnostic Proteomics: Serum Proteomic Patterns for the Detection of Early Stage Cancers

    Directory of Open Access Journals (Sweden)

    Li-Rong Yu

    2004-01-01

    Full Text Available The ability to interrogate thousands of proteins found in complex biological samples using proteomic technologies has brought the hope of discovering novel disease-specific biomarkers. While most proteomic technologies used to discover diagnostic biomarkers are quite sophisticated, "proteomic pattern analysis" has emerged as a simple, yet potentially revolutionary, method for the early diagnosis of diseases. Utilizing this technology, hundreds of clinical samples can be analyzed per day and several preliminary studies suggest proteomic pattern analysis has the potential to be a novel, highly sensitive diagnostic tool for the early detection of cancer.

  8. Detecting and isolating faults of an air-handling unit using on- line diagnostic tests

    Energy Technology Data Exchange (ETDEWEB)

    Pakanen, J. [VTT Building Technology, Espoo (Finland). Building Services and Fire Technology

    1996-12-31

    On-line diagnostic testing is one choice, when practical and robust fault detection and isolation methods are considered for automated processes. Performing a test means exciting a process by means of prescribed input signals, supervising responses and comparing results with a process model. An on-line diagnostic test is repeated similarly every time, in similar process conditions, making modelling an uncomplicated task. Fault detection is a direct consequence of the comparison, but fault isolation is based on elementary constraints, decomposed from the process model. A rough description of a fault can be achieved by heuristic reasoning, which enables application of the method in practice. A more specified fault description is accomplished by learning from old solutions. The reasoner accumulates information making decisions of the classifier gradually more precise through acquired experience. The method is best for successive installations, in which knowledge can be cumulated. On-line diagnostic tests are generic in character, but in this paper they are configured for an air handling unit of an office building and applied in its preheating subprocess. The paper presents the development, simulation and field tests of the fault detection and isolation method and its configuration as a part of a diagnostic system. (orig.) (35 refs.)

  9. Fuzzy-Expert Diagnostics for Detecting and Locating Internal Faults in Three Phase Induction Motors

    Institute of Scientific and Technical Information of China (English)

    DONG Mingchui; CHEANG Takson; SEKAR Booma Devi; CHAN Sileong

    2008-01-01

    Internal faults in three phase induction motors can result in serious performance degradation and eventual system failures if not properly detected and treated in time. Artificial intelligence techniques, the core of soft-computing, have numerous advantages over conventional fault diagnostic approaches; therefore, a soft-computing system was developed to detect and diagnose electric motor faults. The fault diagnostic system for three-phase induction motors samples the fault symptoms and then uses a fuzzy-expert forward inference model to identify the fault. This paper describes how to define the membership functions and fuzzy sets based on the fault symptoms and how to construct the hierarchical fuzzy inference nets with the propagation of probabilities concerning the uncertainty of faults. The designed hierarchical fuzzy inference nets efficiently detect and diagnose the fault type and exact location in a three phase induction motor. The validity and effectiveness of this approach is clearly shown from obtained testing results.

  10. Gold nanodome-patterned microchips for intracellular surface-enhanced Raman spectroscopy.

    Science.gov (United States)

    Wuytens, Pieter C; Subramanian, Ananth Z; De Vos, Winnok H; Skirtach, Andre G; Baets, Roel

    2015-12-21

    While top-down substrates for surface-enhanced Raman spectroscopy (SERS) offer outstanding control and reproducibility of the gold nanopatterns and their related localized surface plasmon resonance, intracellular SERS experiments heavily rely on gold nanoparticles. These nanoparticles often result in varying and uncontrollable enhancement factors. Here we demonstrate the use of top-down gold-nanostructured microchips for intracellular sensing. We develop a tunable and reproducible fabrication scheme for these microchips. Furthermore we observe the intracellular uptake of these structures, and find no immediate influence on cell viability. Finally, we perform a proof-of-concept intracellular SERS experiment by the label-free detection of extraneous molecules. By bringing top-down SERS substrates to the intracellular world, we set an important step towards time-dependent and quantitative intracellular SERS.

  11. A new soft lithographic route for the facile fabrication of hydrophilic sandwich microchips.

    Science.gov (United States)

    Li, Li; Bi, Xiaodong; Yu, Jianzhao; Ren, Carolyn L; Liu, Zhen

    2012-08-01

    Manufacturing materials are an essential element for the fabrication of microfluidic chips. PDMS, the most widely used polymeric material, is associated with apparent disadvantages such as hydrophobic nature, while other materials also suffer from some limitations. In this paper, a new soft lithographic route was proposed for the facile manufacturing of hydrophilic sandwich microchips, using bisphenol A based epoxy acrylate (BABEA) as a new patterning material. The BABEA copolymers are hydrophilic, highly transparent in visible range while highly untransparent when the wavelength is less than 290 nm, and of high replication fidelity. By combining with appropriate monomers, including glycidyl methacrylate, methylmethacrylate, and acrylic acid, the copolymers contain active functional groups, which allows for easy postmodification for desirable functional units. A fabrication procedure was proposed for manufacturing hybrid quartz/BABEA copolymer/quartz microchips. In the procedure, no micromachining equipments, wet etching, or imprinting techniques were involved, making the fabrication approach applicable in ordinary chemistry laboratories. The performance of the prepared microchips was demonstrated in terms of CIEF with UV-whole channel imaging detection. The hydrophilic microchannel ensures stable focusing while the polymeric middle layer acts as a perfectly aligned optical slit for whole channel UV absorbance detection.

  12. Sensitive DNA detection and SNP discrimination using ultrabright SERS nanorattles and magnetic beads for malaria diagnostics.

    Science.gov (United States)

    Ngo, Hoan T; Gandra, Naveen; Fales, Andrew M; Taylor, Steve M; Vo-Dinh, Tuan

    2016-07-15

    One of the major obstacles to implement nucleic acid-based molecular diagnostics at the point-of-care (POC) and in resource-limited settings is the lack of sensitive and practical DNA detection methods that can be seamlessly integrated into portable platforms. Herein we present a sensitive yet simple DNA detection method using a surface-enhanced Raman scattering (SERS) nanoplatform: the ultrabright SERS nanorattle. The method, referred to as the nanorattle-based method, involves sandwich hybridization of magnetic beads that are loaded with capture probes, target sequences, and ultrabright SERS nanorattles that are loaded with reporter probes. Upon hybridization, a magnet was applied to concentrate the hybridization sandwiches at a detection spot for SERS measurements. The ultrabright SERS nanorattles, composed of a core and a shell with resonance Raman reporters loaded in the gap space between the core and the shell, serve as SERS tags for signal detection. Using this method, a specific DNA sequence of the malaria parasite Plasmodium falciparum could be detected with a detection limit of approximately 100 attomoles. Single nucleotide polymorphism (SNP) discrimination of wild type malaria DNA and mutant malaria DNA, which confers resistance to artemisinin drugs, was also demonstrated. These test models demonstrate the molecular diagnostic potential of the nanorattle-based method to both detect and genotype infectious pathogens. Furthermore, the method's simplicity makes it a suitable candidate for integration into portable platforms for POC and in resource-limited settings applications.

  13. Diagnostic tests for the detection of human papillomavirus-associated cervical lesions.

    Science.gov (United States)

    Reuschenbach, Miriam; von Knebel Doeberitz, Magnus

    2013-01-01

    Current diagnostic approaches for primary cervical cancer screening, work-up of equivocal or positive screening results or follow- up after treatment of precancerous lesions primarily rely on the morphologic interpretation of squamous epithelial cells (Pap cytology), in some setting accompanied by the detection of human papillomavirus DNA and have largely contributed to remarkable reduction of disease incidence in countries with implemented screening programs. However, these approaches are limited by a poor sensitivity and reproducibility of Pap cytology and low specificity for high grade cervical intraepithelial neoplasia of HPV DNA detection assays. Early detection might be improved by complementing or even replacing these tests by markers which are more directly related to molecular events triggering HPV-induced carcinogenesis and thereby might deliver more accurate diagnostic performance. The delineation of molecular changes which occur during different stages of HPV infections and the identification of changes which induce neoplastic alterations allow for the detection of markers that specifically highlight the transforming stage of the infection where viral oncogenes are overexpressed and therefore allow for a more specific diagnosis of lesions that require treatment. The evaluation of such markers in clinical studies revealed that some indeed show an improved diagnostic performance compared to Pap cytology or HPV DNA tests only.

  14. Photopatterning of Hydrogel Microarrays in Closed Microchips.

    Science.gov (United States)

    Gumuscu, Burcu; Bomer, Johan G; van den Berg, Albert; Eijkel, Jan C T

    2015-12-14

    To date, optical lithography has been extensively used for in situ patterning of hydrogel structures in a scale range from hundreds of microns to a few millimeters. The two main limitations which prevent smaller feature sizes of hydrogel structures are (1) the upper glass layer of a microchip maintains a large spacing (typically 525 μm) between the photomask and hydrogel precursor, leading to diffraction of UV light at the edges of mask patterns, (2) diffusion of free radicals and monomers results in irregular polymerization near the illumination interface. In this work, we present a simple approach to enable the use of optical lithography to fabricate hydrogel arrays with a minimum feature size of 4 μm inside closed microchips. To achieve this, we combined two different techniques. First, the upper glass layer of the microchip was thinned by mechanical polishing to reduce the spacing between the photomask and hydrogel precursor, and thereby the diffraction of UV light at the edges of mask patterns. The polishing process reduces the upper layer thickness from ∼525 to ∼100 μm, and the mean surface roughness from 20 to 3 nm. Second, we developed an intermittent illumination technique consisting of short illumination periods followed by relatively longer dark periods, which decrease the diffusion of monomers. Combination of these two methods allows for fabrication of 0.4 × 10(6) sub-10 μm sized hydrogel patterns over large areas (cm(2)) with high reproducibility (∼98.5% patterning success). The patterning method is tested with two different types of photopolymerizing hydrogels: polyacrylamide and polyethylene glycol diacrylate. This method enables in situ fabrication of well-defined hydrogel patterns and presents a simple approach to fabricate 3-D hydrogel matrices for biomolecule separation, biosensing, tissue engineering, and immobilized protein microarray applications.

  15. Diagnostic accuracy of xpert test in tuberculosis detection: A systematic review and meta-analysis

    Directory of Open Access Journals (Sweden)

    Ravdeep Kaur

    2016-01-01

    Full Text Available Background: World Health Organization (WHO recommends the use of Xpert MTB/RIF assay for rapid diagnosis of tuberculosis (TB and detection of rifampicin resistance. This systematic review was done to know about the diagnostic accuracy and cost-effectiveness of the Xpert MTB/RIF assay. Methods: A systematic literature search was conducted in following databases: Cochrane Central Register of Controlled Trials and Cochrane Database of Systematic Reviews, MEDLINE, PUBMED, Scopus, Science Direct and Google Scholar for relevant studies for studies published between 2010 and December 2014. Studies given in the systematic reviews were accessed separately and used for analysis. Selection of studies, data extraction and assessment of quality of included studies was performed independently by two reviewers. Studies evaluating the diagnostic accuracy of Xpert MTB/RIF assay among adult or predominantly adult patients (≥14 years, presumed to have pulmonary TB with or without HIV infection were included in the review. Also, studies that had assessed the diagnostic accuracy of Xpert MTB/RIF assay using sputum and other respiratory specimens were included. Results: The included studies had a low risk of any form of bias, showing that findings are of high scientific validity and credibility. Quantitative analysis of 37 included studies shows that Xpert MTB/RIF is an accurate diagnostic test for TB and detection of rifampicin resistance. Conclusion: Xpert MTB/RIF assay is a robust, sensitive and specific test for accurate diagnosis of tuberculosis as compared to conventional tests like culture and microscopic examination.

  16. Diagnostic Accuracy of Xpert Test in Tuberculosis Detection: A Systematic Review and Meta-analysis

    Science.gov (United States)

    Kaur, Ravdeep; Kachroo, Kavita; Sharma, Jitendar Kumar; Vatturi, Satyanarayana Murthy; Dang, Amit

    2016-01-01

    Background: World Health Organization (WHO) recommends the use of Xpert MTB/RIF assay for rapid diagnosis of tuberculosis (TB) and detection of rifampicin resistance. This systematic review was done to know about the diagnostic accuracy and cost-effectiveness of the Xpert MTB/RIF assay. Methods: A systematic literature search was conducted in following databases: Cochrane Central Register of Controlled Trials and Cochrane Database of Systematic Reviews, MEDLINE, PUBMED, Scopus, Science Direct and Google Scholar for relevant studies for studies published between 2010 and December 2014. Studies given in the systematic reviews were accessed separately and used for analysis. Selection of studies, data extraction and assessment of quality of included studies was performed independently by two reviewers. Studies evaluating the diagnostic accuracy of Xpert MTB/RIF assay among adult or predominantly adult patients (≥14 years), presumed to have pulmonary TB with or without HIV infection were included in the review. Also, studies that had assessed the diagnostic accuracy of Xpert MTB/RIF assay using sputum and other respiratory specimens were included. Results: The included studies had a low risk of any form of bias, showing that findings are of high scientific validity and credibility. Quantitative analysis of 37 included studies shows that Xpert MTB/RIF is an accurate diagnostic test for TB and detection of rifampicin resistance. Conclusion: Xpert MTB/RIF assay is a robust, sensitive and specific test for accurate diagnosis of tuberculosis as compared to conventional tests like culture and microscopic examination. PMID:27013842

  17. Electrochemical methods in conjunction with capillary and microchip electrophoresis.

    Science.gov (United States)

    Mark, Jonas J P; Scholz, Rebekka; Matysik, Frank-Michael

    2012-12-01

    Electromigrative techniques such as capillary and microchip electrophoresis (CE and MCE) are inherently associated with various electrochemical phenomena. The electrolytic processes occurring in the buffer reservoirs have to be considered for a proper design of miniaturized electrophoretic systems and a suitable selection of buffer composition. In addition, the control of the electroosmotic flow plays a crucial role for the optimization of CE/MCE separations. Electroanalytical methods have significant importance in the field of detection in conjunction with CE/MCE. At present, amperometric detection and contactless conductivity detection are the predominating electrochemical detection methods for CE/MCE. This paper reviews the most recent trends in the field of electrochemical detection coupled to CE/MCE. The emphasis is on methodical developments and new applications that have been published over the past five years. A rather new way for the implementation of electrochemical methods into CE systems is the concept of electrochemically assisted injection which involves the electrochemical conversions of analytes during the injection step. This approach is particularly attractive in hyphenation to mass spectrometry (MS) as it widens the range of CE-MS applications. An overview of recent developments of electrochemically assisted injection coupled to CE is presented.

  18. Oligonucleotide-based biosensors for in vitro diagnostics and environmental hazard detection.

    Science.gov (United States)

    Jung, Il Young; Lee, Eun Hee; Suh, Ah Young; Lee, Seung Jin; Lee, Hyukjin

    2016-04-01

    Oligonucleotide-based biosensors have drawn much attention because of their broad applications in in vitro diagnostics and environmental hazard detection. They are particularly of interest to many researchers because of their high specificity as well as excellent sensitivity. Recently, oligonucleotide-based biosensors have been used to achieve not only genetic detection of targets but also the detection of small molecules, peptides, and proteins. This has further broadened the applications of these sensors in the medical and health care industry. In this review, we highlight various examples of oligonucleotide-based biosensors for the detection of diseases, drugs, and environmentally hazardous chemicals. Each example is provided with detailed schematics of the detection mechanism in addition to the supporting experimental results. Furthermore, future perspectives and new challenges in oligonucleotide-based biosensors are discussed.

  19. Diagnostic accuracy of cone beam computed tomography in detection of simulated mandibular condyle erosions

    OpenAIRE

    Shahriar Shahab; Nafiseh Nikkerdar; Maryam Goodarzi; Amin Golshah; Sanaz Sharifi Shooshtari

    2015-01-01

    Introduction: To determine the diagnostic accuracy of cone beam computed tomography (CBCT) in the detection of simulated mandibular condyle erosions. Materials and Methods: Seventeen dry human mandibles were used in this in vitro study. NewTom VG CBCT scanner (New Tom VG, Verona, Veneto region, Italy) was used for the condyles imaging (pre-erosion and post-erosion image). Thirty three lesions were created on the superior (11 cases), anterior (11 cases), and posterior surfaces (11 cases) o...

  20. Diagnostic methods to cutaneous leishmaniasis detection in domestic dogs and cats*

    OpenAIRE

    Trevisan, Daliah Alves Coelho; Lonardoni, Maria Valdrinez Campana; DEMARCHI, Izabel Galhardo

    2015-01-01

    Cutaneous leishmaniasis is caused by different species of Leishmania. In domestic animals such as dogs and cats, the diagnostic consists of clinical, epidemiological and serological tests, which changes among countries all around the world. Because of this diversity in the methods selected, we propose this systematic literature review to identify the methods of laboratory diagnosis used to detect cutaneous leishmaniasis in domestic dogs and cats in the Americas. Articles published in the last...

  1. Diagnostic methods to cutaneous leishmaniasis detection in domestic dogs and cats

    OpenAIRE

    Trevisan, Daliah Alves Coelho; Lonardoni,Maria Valdrinez Campana; Demarchi,Izabel Galhardo

    2015-01-01

    Abstract: Cutaneous leishmaniasis is caused by different species of Leishmania. In domestic animals such as dogs and cats, the diagnostic consists of clinical, epidemiological and serological tests, which changes among countries all around the world. Because of this diversity in the methods selected, we propose this systematic literature review to identify the methods of laboratory diagnosis used to detect cutaneous leishmaniasis in domestic dogs and cats in the Americas. Articles published i...

  2. Combination of Culture, Antigen and Toxin Detection, and Cytotoxin Neutralization Assay for Optimal Clostridium difficile Diagnostic Testing

    Directory of Open Access Journals (Sweden)

    Michelle J Alfa

    2013-01-01

    Full Text Available BACKGROUND: There has been a growing interest in developing an appropriate laboratory diagnostic algorithm for Clostridium difficile, mainly as a result of increases in both the number and severity of cases of C difficile infection in the past decade. A C difficile diagnostic algorithm is necessary because diagnostic kits, mostly for the detection of toxins A and B or glutamate dehydrogenase (GDH antigen, are not sufficient as stand-alone assays for optimal diagnosis of C difficile infection. In addition, conventional reference methods for C difficile detection (eg, toxigenic culture and cytotoxin neutralization [CTN] assays are not routinely practiced in diagnostic laboratory settings.

  3. 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.

  4. Studies on Enzyme Kinetics by Microchip and Related Techniques

    Institute of Scientific and Technical Information of China (English)

    James J. Bao; WANG Huai-Feng; ZHOU Da-Wei; Ken R. Wehmeyer

    2004-01-01

    Both conventional and microchip-based capillary electrophoresis(CE) technologies have been used for the analysis of enzymes. Practical procedures of using CE to determine the Km and Vmax values of an enzyme have been developed. By studying the inhibition to the enzyme, it is possible to select a suitable drug candidate. When compared with the conventional CE method, single lane microchip-based method can improve the speed for the assay three times. By using multiple lane-based microchip, the speed can be further increased.

  5. Diagnostic Performance of CT Colonography for the Detection of Colorectal Polyps

    Energy Technology Data Exchange (ETDEWEB)

    Yun, Ji Young; Ro, Hee Jeong; Choi, Jung Bin; Chung, Ji Eun; Kim, Yong Jin; Suh, Won Hyuck; Lee, Jong Kyun [Song-Do Hospital, Seoul (Korea, Republic of); Park, Jong Beom [East- West Neo Medical Center, Kyung Hee University, Seoul (Korea, Republic of)

    2007-12-15

    To investigate the diagnostic value of CT colonography for the detection of colorectal polyps. From December 2004 to December 2005, 399 patients underwent CT colonography and follow-up conventional colonoscopy. We excluded cases of advanced colorectal cancer. We retrospectively analyzed the CT colonography findings and follow-up conventional colonoscopy findings of 113 patients who had polyps more than 6 mm in diameter. Radiologists using 3D and 2D computer generated displays interpreted the CT colonography images. The colonoscopists were aware of the CT colonography findings before the procedure. CT colonography detected 132 polyps in 107 of the 113 patients and conventional colonoscopy detected 114 colorectal polyps more than 6 mm in diameter in 87 of the 113 patients. The sensitivity of CT colonography analyzed per polyp was 91% (41/45) for polyps more than 10 mm in diameter and 89% (101/114) for polyps more than 6 mm in diameter. Thirteen polyps were missed by CT colonography and were detected on follow-up conventional colonoscopy. CT colonography is a sensitive diagnostic tool for the detection of colorectal polyps and adequate bowel preparation, optimal bowel distention and clinical experience are needed to reduce the rate of missing appropriate lesions.

  6. Low-cost fabrication of poly(methyl methacrylate) microchips using disposable gelatin gel templates.

    Science.gov (United States)

    Chen, Zhi; Yu, Zhengyin; Chen, Gang

    2010-06-15

    A simple method based on disposable gelatin gel templates has been developed for the low-cost fabrication of poly(methyl methacrylate) (PMMA) microfluidic chips. Gelatin was dissolved in glycerol aqueous solution under heat to prepare a thermally reversible impression material. The molten gel was then sandwiched between a glass plate and a SU-8 template bearing negative relief of microstructure. After cooling, the negative SU-8 template could be easily separated from the solidified gelatin gel and a layer of gelatin template bearing positive relief of the microstructure was left on the glass plate. Subsequently, prepolymerized methyl methacrylate molding solution containing a UV-initiator was sandwiched between the gel template and a PMMA plate and was allowed to polymerize under UV light to fabricate PMMA channel plate at room temperature. Complete microchips could be obtained by bonding the channel plates with covers using plasticizer-assisted thermal bonding at 90 degrees C. Gelatin gel template can be mass-produced and will find application in the mass production of PMMA microchips at low cost. The prepared microfluidic microchips have been successfully employed in the capillary electrophoresis analysis of several ions in connection with contactless conductivity detection.

  7. Incremental diagnostic value of combined quantitative and qualitative parameters of magnetocardiography to detect coronary artery disease.

    Science.gov (United States)

    Shin, Eun-Seok; Lam, Yat-Yin; Her, Ae-Young; Brachmann, Johannes; Jung, Friedrich; Park, Jai-Wun

    2017-02-01

    Magnetocardiography (MCG) has been proposed as a non-invasive and functional technique with high accuracy for diagnosis of myocardial ischemia. This study sought to investigate the incremental diagnostic value of combined quantitative and qualitative parameters of MCG to detect coronary artery disease (CAD). Ninety six patients with suspected CAD who underwent coronary angiography were enrolled in the analysis to test the diagnostic accuracy of 2 MCG parameters (a quantitative parameter of the percent change of ST-segment fluctuation score and a qualitative parameter of non-dipole phenomenon). The best cut-off value for the percent change of ST-segment fluctuation score was -51.0%. The accuracy, sensitivity, specificity, positive predictive value, and negative predictive value were 78.1, 73.9, 82.0, 79.1, and 77.4, in the percent change of ST-segment fluctuation score and 86.5, 84.8, 88.0, 86.7, and 86.3 in non-dipole phenomenon. The area under the curve of receiver-operating characteristics was 0.79 for the percent change of ST-segment fluctuation score and 0.86 for non-dipole phenomenon (pQualitative assessment of non-dipole phenomenon has a better diagnostic value than the quantitative parameter of percent change of ST-segment fluctuation score in the detection of significant CAD. Furthermore, this study found that the incorporation of non-dipole phenomenon into the percent change of ST-segment fluctuation score significantly improved the diagnostic performance of CAD detection. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  8. A microcosting study of diagnostic tests for the detection of coronary artery disease in the Netherlands

    Energy Technology Data Exchange (ETDEWEB)

    Tan, S.S. [Erasmus MC University Medical Center, Institute for Medical Technology Assessment, P.O. Box 1738, 3000 DR Rotterdam (Netherlands)], E-mail: s.s.tan@erasmusmc.nl; Oppe, M. [Erasmus MC University Medical Center, Institute for Medical Technology Assessment, P.O. Box 1738, 3000 DR Rotterdam (Netherlands)], E-mail: m.oppe@erasmusmc.nl; Zoet-Nugteren, S.K. [Ikazia Hospital Rotterdam, Department of Cardiology, P.O. Box 5009, 3008 AA Rotterdam (Netherlands)], E-mail: sk.zoet@ikazia.nl; Niezen, R.A. [Medical Center Rijnmond-Zuid, Department of Radiology, P.O. Box 9100, 3007 AC Rotterdam (Netherlands)], E-mail: niezenr@mcrz.nl; Kofflard, M.J.M. [Albert Schweitzer Hospital, Department of Cardiology, P.O. Box 444, 3300 AK Dordrecht (Netherlands)], E-mail: m.j.m.kofflard@asz.nl; Ten Cate, F.J. [Erasmus MC University Medical Center, Department of Cardiology, P.O. Box 2040, 3000 CA Rotterdam (Netherlands)], E-mail: f.j.tencate@erasmusmc.nl; Roijen, L. Hakkaart-van [Erasmus MC University Medical Center, Institute for Medical Technology Assessment, P.O. Box 1738, 3000 DR Rotterdam (Netherlands)], E-mail: l.hakkaart-vanroijen@erasmusmc.nl

    2009-10-15

    Objective: The primary aim of the present study was to calculate the actual costs of four diagnostic tests for the detection of coronary artery disease in the Netherlands using a microcosting methodology. As a secondary objective, the cost effectiveness of eight diagnostic strategies was examined, using microcosting and reimbursement fees subsequently as the cost estimate. Design: A multicenter, retrospective cost analysis from a hospital perspective. Setting: The study was conducted in three general hospitals in the Netherlands for 2006. Interventions: Exercise electrocardiography (exECG), stress echocardiography (sECHO), single-photon emission computed tomography (SPECT) and coronary angiography (CA). Results: The actual costs of exECG, sECHO, SPECT and CA were Euro 33, 216, 614 and 1300 respectively. For all diagnostic tests, labour and indirect cost components (overheads and capital) together accounted for over 75% of the total costs. Consumables played a relatively important role in SPECT (14%). Hotel and nutrition were only applicable to SPECT and CA. Diagnostic services were solely performed for CA, but their costs were negligible (2%). Using microcosting estimates, exECG-sECHO-SPECT-CA was the most and CA the least cost effective strategy ( Euro 397 and 1302 per accurately diagnosed patient). Using reimbursement fees, exECG-sECHO-CA was most and SPECT-CA least cost effective ( Euro 147 and 567 per accurately diagnosed patient). Conclusions: The use of microcosting estimates instead of reimbursement fees led to different conclusions regarding the relative cost effectiveness of alternative strategies.

  9. Evaluation of diagnostic ability of CCD digital radiography in the detection of incipient dental caries

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Wan; Lee, Byung Do [Wonkwang University College of Medicine, Iksan (Korea, Republic of)

    2003-03-15

    The purpose of this experiment was to evaluate the diagnostic ability of a CCD-based digital system (CDX-2000HQ) in the detection of incipient dental caries. 93 extracted human teeth with sound proximal surfaces and interproximal artificial cavities were radiographed using 4 imaging methods. Automatically processed No.2 Insight film (Eastman Kodak Co., U.S.A.) was used for conventional radiography, scanned images of conventional radiograms for indirect digital radiography were used. For the direct digital radiography, the CDX-2000HQ CCD system (Biomedisys Co. Korea) was used. The subtraction images were made from two direct digital images by Sunny program in the CDX-2000HQ system. Two radiologists and three endodontists examined the presence of lesions using a five-point confidence scale and compared the diagnostic ability by ROC (Receiver Operating Characteristic) analysis and one way ANOVA test. The mean ROC areas of conventional radiography, indirect digital radiography, direct digital radiography, and digital subtraction radiography were 0.9093, 0.9102, 0.9184, and 0.9056, respectively. The diagnostic ability of direct digital radiography was better than the other imaging modalities, but there were no statistical differences among these imaging modalities (p>0.05). These results indicate that new CCD-based digital systems (CDX-2000HQ) have the potential to serve as an alternative to conventional radiography in the detection of incipient dental caries.

  10. Comparison of the diagnostic performance of panoramic and occlusal radiographs in detecting submandibular sialoliths

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jun Ho; Aoki, Eduardo Massaharu; Cortes, Arthur Rodriguez Gonzalez; Arita, Emiko Saito; Abdala, Reinaldo Junior [Dept. of Oral Radiology, School of Dentistry, University of Sao Paulo, Sao Paulo (Brazil); Asaumi, Junichi [Dept. of Oral Radiology, School of Dentistry, University of Okayama, Okayama (Japan)

    2016-06-15

    The aim of this study was to assess and compare the diagnostic performance of panoramic and occlusal radiographs in detecting submandibular sialoliths. A total of 40 patients (20 cases and 20 controls) were included in this retrospective study. Cases were defined as subjects with a submandibular sialolith confirmed by computed tomography (CT), whereas controls did not have any submandibular calcifications. Three observers with different expertise levels assessed panoramic and occlusal radiographs of all subjects for the presence of sialoliths. Intraobserver and interobserver agreement were assessed using the kappa test. Sensitivity, specificity, accuracy, positive and negative predictive values, and the diagnostic odds ratio of panoramic and occlusal radiographs in screening for submandibular sialoliths were calculated for each observer. The sensitivity and specificity values for occlusal and panoramic radiographs all ranged from 80% to 100%. The lowest values of sensitivity and specificity observed among the observers were 82.6% and 80%, respectively (P=0.001). Intraobserver and interobserver agreement were higher for occlusal radiographs than for panoramic radiographs, although panoramic radiographs demonstrated a higher overall accuracy. Both panoramic and occlusal radiographic techniques displayed satisfactory diagnostic performance and should be considered before using a CT scan to detect submandibular sialoliths.

  11. Diagnostic Value of CT for the Detection of Cervical Lymph Node Metastases in Papillary Thyroid Carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Son, Kyu Ri [Boramae Medical Center, Seoul (Korea, Republic of); Na, Dong Gyu; Chang, Kee Hyun [Seoul National University Hospital, Seoul (Korea, Republic of)

    2009-06-15

    To determine the diagnostic accuracy of CT for the detection of cervical lymph node metastases in patients with papillary thyroid carcinoma (PTC). Two hundred twelve consecutive patients with surgically proven PTC were included in this study. CT images were retrospectively evaluated to determine the presence of a node metastasis using morphologic CT criteria (at least one of the following: strong nodal enhancement without hilar vessel enhancement, heterogeneous enhancement, calcification, and cystic change). The diagnostic accuracy of CT for the diagnosis of a metastatic lymph node was assessed using a level-bylevel analysis. The accuracy of the CT finding for strong nodal enhancement was greater than the other morphologic CT criteria (81.6% and 74.5-78.5%, respectively). The sensitivity, specificity, and accuracy were 64.4%, 91.4%, and 84.3% by the morphologic CT criteria, and were 34.6%, 93.9%, and 78.2% by size criteria, respectively. The morphologic CT criteria are more accurate than the size criteria in the detection of cervical lymph node metastases in patients with papillary thyroid carcinoma; and, strong nodal enhancement on a CT scan is the most important factor for its diagnostic accuracy.

  12. Improving detection of avian malaria from host blood: a step towards a standardised protocol for diagnostics.

    Science.gov (United States)

    Niebuhr, Chris N; Blasco-Costa, Isabel

    2016-10-01

    Avian malaria, caused by Plasmodium spp., has been linked to the mortality and population-level declines in native birds in some regions. While molecular diagnostic methods have greatly improved our ability to detect infections of both human and bird malaria, failing to identify false negatives remains an important handicap, particularly for avian malaria due to host DNA presence in the bird blood cells. In an attempt to improve the accuracy of diagnostics by PCR, we evaluated the performance of a commercial silica-membrane-based DNA extraction kit by modifying the protocol with four unpooled elution volume alternatives. Our results suggest that the best template is the DNA extract obtained from the second eluate of a first 50 μL elution step. In one case, the only band visible was from this second eluate and, thus, may not have been identified as positive for Plasmodium spp. if a different elution protocol had been followed. Our results are likely explained by the concept of size exclusion chromatography by which particles of different sizes will elute at different rates. Overall, first elution templates may consist of a lower ratio of parasite to host DNA, while second eluates may contain a higher parasite to host DNA ratio. A low ratio of parasite to host DNA is a concern in detecting chronic infections, in which birds typically carry low levels of parasitemia, making accurate diagnostics imperative when identifying reservoirs of disease that could lead to spillback events.

  13. A smartphone-based diagnostic platform for rapid detection of Zika, chikungunya, and dengue viruses

    Science.gov (United States)

    Priye, Aashish; Bird, Sara W.; Light, Yooli K.; Ball, Cameron S.; Negrete, Oscar A.; Meagher, Robert J.

    2017-01-01

    Current multiplexed diagnostics for Zika, dengue, and chikungunya viruses are situated outside the intersection of affordability, high performance, and suitability for use at the point-of-care in resource-limited settings. Consequently, insufficient diagnostic capabilities are a key limitation facing current Zika outbreak management strategies. Here we demonstrate highly sensitive and specific detection of Zika, chikungunya, and dengue viruses by coupling reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with our recently developed quenching of unincorporated amplification signal reporters (QUASR) technique. We conduct reactions in a simple, inexpensive and portable “LAMP box” supplemented with a consumer class smartphone. The entire assembly can be powered by a 5 V USB source such as a USB power bank or solar panel. Our smartphone employs a novel algorithm utilizing chromaticity to analyze fluorescence signals, which improves the discrimination of positive/negative signals by 5-fold when compared to detection with traditional RGB intensity sensors or the naked eye. The ability to detect ZIKV directly from crude human sample matrices (blood, urine, and saliva) demonstrates our device’s utility for widespread clinical deployment. Together, these advances enable our system to host the key components necessary to expand the use of nucleic acid amplification-based detection assays towards point-of-care settings where they are needed most. PMID:28317856

  14. Application of hybridization control probe to increase accuracy on ligation detection or minisequencing diagnostic microarrays

    Directory of Open Access Journals (Sweden)

    Hultman Jenni

    2009-12-01

    Full Text Available Abstract Background Nucleic acid detection based on ligation reaction or single nucleotide extension of ssDNA probes followed by tag microarray hybridization provides an accurate and sensitive detection tool for various diagnostic purposes. Since microarray quality is crucial for reliable detection, these methods can benefit from correcting for microarray artefacts using specifically adapted techniques. Findings Here we demonstrate the application of a per-spot hybridization control oligonucleotide probe and a novel way of computing normalization for tag array data. The method takes into account the absolute value of the detection probe signal and the variability in the control probe signal to significantly alleviate problems caused by artefacts and noise on low quality microarrays. Conclusions Diagnostic microarray platforms require experimental and computational tools to enable efficient correction of array artefacts. The techniques presented here improve the signal to noise ratio and help in determining true positives with better statistical significance and in allowing the use of arrays with poor quality that would otherwise be discarded.

  15. A smartphone-based diagnostic platform for rapid detection of Zika, chikungunya, and dengue viruses.

    Science.gov (United States)

    Priye, Aashish; Bird, Sara W; Light, Yooli K; Ball, Cameron S; Negrete, Oscar A; Meagher, Robert J

    2017-03-20

    Current multiplexed diagnostics for Zika, dengue, and chikungunya viruses are situated outside the intersection of affordability, high performance, and suitability for use at the point-of-care in resource-limited settings. Consequently, insufficient diagnostic capabilities are a key limitation facing current Zika outbreak management strategies. Here we demonstrate highly sensitive and specific detection of Zika, chikungunya, and dengue viruses by coupling reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with our recently developed quenching of unincorporated amplification signal reporters (QUASR) technique. We conduct reactions in a simple, inexpensive and portable "LAMP box" supplemented with a consumer class smartphone. The entire assembly can be powered by a 5 V USB source such as a USB power bank or solar panel. Our smartphone employs a novel algorithm utilizing chromaticity to analyze fluorescence signals, which improves the discrimination of positive/negative signals by 5-fold when compared to detection with traditional RGB intensity sensors or the naked eye. The ability to detect ZIKV directly from crude human sample matrices (blood, urine, and saliva) demonstrates our device's utility for widespread clinical deployment. Together, these advances enable our system to host the key components necessary to expand the use of nucleic acid amplification-based detection assays towards point-of-care settings where they are needed most.

  16. Diagnostic Yield of High-Resolution Breast Sonography in Detecting Microcalcifications Compared to Mammography

    Directory of Open Access Journals (Sweden)

    N Ahmadinejad

    2009-08-01

    Full Text Available Background/Objective: Mammography remains the most suitable screening test in detecting microcalcifications as the earliest manifestation of breast malignancy. By means of highfrequency transducers yielding high-resolution breast imaging, some researchers have reported that ultrasonography is capable of depicting microcalcifications in the breast tissue. Therefore, this study has been designed to compare the diagnostic yield of high-resolution"nbreast ultrasonography (HRS versus conventional mammography."nPatients and Methods: Seventy-four consecutive patients who had breast microcalcifications (hyperdense foci < 0.5mm according to standard mammograms, without a prior history of breast disease, surgery, biopsy, chest wall radiation or systemic chemotherapy were enrolled. Considering mammograms as a reference, 46 patients without a mass, voluntarily underwent high-resolution bilateral breast ultrasonography."nResults: The mean age was 50.7±10 years (range, 35-85 years. The upper outer quadrant of the breast was the commonest place where microcalcifications were detected (36.9%. A relative frequency of 45.7% was reported for microcalcifications with breast imaging reporting"nand data system (BIRADS score 3. An overall 82.6% diagnostic yield was discovered for HRS in detecting microcalcifications; it detected all microcalcifications with BIRADS score 4 and 5, but 57.1% and 90.5% of microcalcifications with BIRADS score 2 and 3, respectively. Cluster microcalcification was the most common pattern (43.5%."nConclusion: Considering the 82.6% diagnostic yield of HRS compared to mammography, it can be proposed as the surrogate modality in locating microcalcifications in procedures such as biopsies and hook-wiring, with the advantage of reducing radiation exposure. HRS may be the future screening modality as a result of feasibility, safety, compliance and accuracy.

  17. Antigen Detection in the Diagnosis of Histoplasmosis: A Meta-analysis of Diagnostic Performance.

    Science.gov (United States)

    Fandiño-Devia, Estefanía; Rodríguez-Echeverri, Carolina; Cardona-Arias, Jaiberth; Gonzalez, Angel

    2016-04-01

    We performed a meta-analysis of diagnostic data to evaluate the performance of Histoplasma antigen detection tests for diagnosing histoplasmosis. We included all studies involving human subjects that assessed the performance of any antigen detection test for histoplasmosis in urine or serum by carrying out an exhaustive and reproducible search of the literature between 1980 and 2014 from four databases. Quality of the articles was assessed, and meta-analysis was performed under the random effects model, calculating sensitivity, specificity, likelihood and odds ratios, and ROC curve using Meta-DiSc(es). Nine out of a total of 23 studies met strict quality criteria and were therefore included. The overall sensitivity for antigen detection in serum and urine was 81% (95% CI 78-83%), while specificity was 99% (95% CI 98-99%). Sensitivity for antigenuria and antigenemia was 79% (95% CI 76-82%) and 82% (95% CI 79-85%), respectively; specificity values were 99% (95% CI 98-100%) in urine and 97% (95% CI 96-98%) in serum. The positive and negative likelihood ratios were 49.5 (95% CI 20.7-118.7) and 0.19 (95% CI 0.14-0.26), respectively, while the diagnostic OR was 362 (95% CI 121.2-1080.3) and area under the curve was 0.99. In conclusion, the performance of Histoplasma antigen detection assay of urine was not significantly different from that of blood, indicating that antigenuria and antigenemia have equal diagnostic value in histoplasmosis.

  18. A real-time PCR diagnostic method for detection of Naegleria fowleri.

    Science.gov (United States)

    Madarová, Lucia; Trnková, Katarína; Feiková, Sona; Klement, Cyril; Obernauerová, Margita

    2010-09-01

    Naegleria fowleri is a free-living amoeba that can cause primary amoebic meningoencephalitis (PAM). While, traditional methods for diagnosing PAM still rely on culture, more current laboratory diagnoses exist based on conventional PCR methods; however, only a few real-time PCR processes have been described as yet. Here, we describe a real-time PCR-based diagnostic method using hybridization fluorescent labelled probes, with a LightCycler instrument and accompanying software (Roche), targeting the Naegleria fowleriMp2Cl5 gene sequence. Using this method, no cross reactivity with other tested epidemiologically relevant prokaryotic and eukaryotic organisms was found. The reaction detection limit was 1 copy of the Mp2Cl5 DNA sequence. This assay could become useful in the rapid laboratory diagnostic assessment of the presence or absence of Naegleria fowleri.

  19. A hydrodynamic microchip for formation of continuous cell chains

    Science.gov (United States)

    Khoshmanesh, Khashayar; Zhang, Wei; Tang, Shi-Yang; Nasabi, Mahyar; Soffe, Rebecca; Tovar-Lopez, Francisco J.; Rajadas, Jayakumar; Mitchell, Arnan

    2014-05-01

    Here, we demonstrate the unique features of a hydrodynamic based microchip for creating continuous chains of model yeast cells. The system consists of a disk shaped microfluidic structure, containing narrow orifices that connect the main channel to an array of spoke channels. Negative pressure provided by a syringe pump draws fluid from the main channel through the narrow orifices. After cleaning process, a thin layer of water is left between the glass substrate and the polydimethylsiloxane microchip, enabling leakage beneath the channel walls. A mechanical clamp is used to adjust the operation of the microchip. Relaxing the clamp allows leakage of liquid beneath the walls in a controllable fashion, leading to formation of a long cell chain evenly distributed along the channel wall. The unique features of the microchip are demonstrated by creating long chains of yeast cells and model 15 μm polystyrene particles along the side wall and analysing the hydrogen peroxide induced death of patterned cells.

  20. Indian microchip for Big Bang research in Geneva

    CERN Multimedia

    Bhabani, Soudhriti

    2007-01-01

    "A premier nuclear physics institute here has come up with India's first indigenously designed microchip that will facilitate research on the Big Bang theory in Geneva's CERN, the world's largest particle physics laboratory." (1 page)

  1. Poly(dimethylsiloxane) based microchip for DNA electrophoresis

    Institute of Scientific and Technical Information of China (English)

    LIU Changchun; CUI Dafu; WANG Li

    2004-01-01

    A novel poly(dimethylsiloxane)(PDMS) -based microchip for DNA separation through electrophoresis has been developed using a micro-electro-mechanical-system(MEMS) technology. Unlike previous hybrid PDMS microchip, one PDMS film is first created on glass support by pressing method in our microchip. Thus, increased band-broadening phenomena, arising from the material nonuniformity at the walls of microchannel, can be avoided in electrophoresis process. A low-viscosity hydroxypropylmethylcellulose-100 (HPMC-100) is used as the separation medium for fluorescent intercalator-labeled double-stranded DNA (dsDNA) fragments. Mannitol is introduced to PDMS-based microchip as a separation medium additive to enhance separation efficiency. At applied electric field strength of 150 V/cm, excellent separations of the PCR marker could be achieved with an effective separation distance of 25mm .

  2. Diagnostic accuracy of cone beam computed tomography in detection of simulated mandibular condyle erosions

    Directory of Open Access Journals (Sweden)

    Shahriar Shahab

    2015-01-01

    Full Text Available Introduction: To determine the diagnostic accuracy of cone beam computed tomography (CBCT in the detection of simulated mandibular condyle erosions. Materials and Methods: Seventeen dry human mandibles were used in this in vitro study. NewTom VG CBCT scanner (New Tom VG, Verona, Veneto region, Italy was used for the condyles imaging (pre-erosion and post-erosion image. Thirty three lesions were created on the superior (11 cases, anterior (11 cases, and posterior surfaces (11 cases of the condyles. The pre- and post-erosion images were randomly presented to two previously calibrated oral and maxillofacial radiologists in order to evaluate the presence of simulated erosions and their position in the condyles using two protocols. In the first protocol, axial and coronal images and in the second protocol, axial, coronal, and sagittal/multiplanar reconstructed (MPR images were used to evaluate the lesions of the samples. Furthermore, the Cochran′s Q test and McNemar and Kappa statistical tests were used to assess the sensitivity, specificity, and accuracy of this study. Results: There was no statistically significant difference between the diagnostic methods and the reference value. There was substantial agreement between the two protocols (Kappa > 0.61. Protocol 2 showed relatively better results than protocol 1 but the difference was not statistically significant (P > 0.05. Sensitivity, specificity, and diagnostic accuracy levels in the erosion imaging were higher in the posterior region of condyle; however, there was no statistically significant difference between the condylar regions (P > 0.05. Conclusion: CBCT had high sensitivity, specificity, and diagnostic accuracy in the detection of simulated mandibular condyle erosions.

  3. Early detection of prostate cancer relapse by biochemistry and diagnostic imaging.

    Science.gov (United States)

    Evangelista, L; Zattoni, F; Rossi, E; Karnes, R J; Lowe, V

    2015-12-01

    Prostate cancer (PCa) is a common malignancy in men associated with an increase in the incidence rate. Radical prostatectomy (RP) or external beam radiotherapy (EBRT) represents the most employed treatments for the local control of disease. However, 10-50% of patients who experienced a recurrence of disease after primary treatments can benefit from salvage or palliative therapies. To date, prostate specific antigen (PSA) is usually used in clinical practice to monitor the status of disease and to early detect the recurrence of PCa. Nevertheless, PSA cannot discriminate the presence of local vs. distant metastatic disease. Circulating tumor cells are considered as a sign of disease widespread, but their correlation with metastatic PCa and local recurrence of disease is still indeterminate. Digital rectal exploration and transrectal ultrasonography are considered the first clinical and diagnostic approach to identify the local recurrence of PCa, but are associated with a low detection rate and low diagnostic accuracies. Conversely, magnetic resonance imaging (MRI) has gained a great importance in this setting of disease, being able to determine the presence of local recurrence with high sensitivity, also in the presence of low serum PSA levels. Lastly, the introduction of positron emission tomography/computed tomography (PET/CT) with radiolabeled choline agents let to improve the management of patients with early recurrence of disease, although its accuracy is linked to the PSA and PSA dynamic values. New radiopharmaceutical agents, like 68Ga-PSMA or 18F-FACBC and others could improve the diagnostic accuracy of PET/CT, but the data is still preliminary. In the present review we will discuss both clinical and diagnostic instrumentations, actually available in clinical practice, able to early identify the presence of recurrent PCa and to differentiate between local and distant relapse of tumor.

  4. Apparatus for Precise Indium-Bump Bonding of Microchips

    Science.gov (United States)

    Wild, Larry; Mulder, Jerry; Alvarado, Nicholas

    2005-01-01

    An improved apparatus has been designed and built for use in precise positioning and pressing of a microchip onto a substrate (which could, optionally, be another microchip) for the purpose of indium-bump bonding. The apparatus (see figure) includes the following: A stereomicroscope, A stage for precise positioning of the microchip in rotation angle (theta) about the nominally vertical pressing axis and in translation along two nominally horizontal coordinate axes (x and y), and An actuator system that causes a bonding tip to press the microchip against the substrate with a precisely controlled force. In operation, the microscope and the stage are used to position the microchip under the bonding tip and to align the indium bumps on the chip and the substrate, then the actuator system is used to apply a prescribed bonding force for a prescribed time. The improved apparatus supplants a partly similar prior apparatus that operated with less precision and repeatability, producing inconsistent and unreliable bonds. Results of the use of the prior apparatus included broken microchips, uneven bonds, and bonds characterized, variously, by overcompression or undercompression. In that apparatus, the bonding force was generated and controlled by use of a micrometer head positioned over the center of a spring-loaded scale, and the force was applied to the microchip via the scale, which was equipped for digital readout of the force. The inconsistency of results was attributed to the following causes: It was not possible to control the bonding force with sufficient precision or repeatability. Particularly troublesome was the inability to control the force at levels less than the weight of 150 g. Excessive compliance in the spring-loaded scale, combined with deviations from parallelarity of the substrate and bonding-tip surfaces, gave rise to nonuniformity in the pressure applied to the microchip, thereby generating excessive stresses and deformations in the microchip. In the

  5. Molecular method for the detection of Andes hantavirus infection: validation for clinical diagnostics

    Science.gov (United States)

    Vial, Cecilia; Martinez-Valdebenito, Constanza; Rios, Susana; Martinez, Jessica; Vial, Pablo; Ferres, Marcela; Rivera, Juan Carlos; Perez, Ruth; Valdivieso, Francisca

    2016-01-01

    Hantavirus Cardiopulmonary Syndrome is a severe disease caused by exposure to New World hantaviruses. Early diagnosis is difficult due to the lack of specific initial symptoms. Anti-hantavirus antibodies are usually negative until late in the febrile prodrome or the beginning of cardiopulmonary phase while Andes hantavirus (ANDV) RNA genome can be detected before symptoms onset. We analyzed the effectiveness of RTqPCR as a diagnostic tool detecting ANDV-Sout genome in peripheral blood cells from 78 confirmed hantavirus patients and 166 negative controls. Our results indicate that RTqPCR had a low detection limit (~10 copies), with a specificity of 100% and a sensitivity of 94.9%. This suggests the potential for establishing RT-qPCR as the assay of choice for early diagnosis, promoting early effective care of patients and improve other important aspects of ANDV infection management, such as compliance of biosafety recommendations for health personnel in order to avoid nosocomial transmission. PMID:26508102

  6. Carbon Nanotube Biosensors for Space Molecule Detection and Clinical Molecular Diagnostics

    Science.gov (United States)

    Han, Jie

    2001-01-01

    Both space molecule detection and clinical molecule diagnostics need to develop ultra sensitive biosensors for detection of less than attomole molecules such as amino acids for DNA. However all the electrode sensor systems including those fabricated from the existing carbon nanotubes, have a background level of nA (nanoAmp). This has limited DNA or other molecule detection to nA level or molecules whose concentration is, much higher than attomole level. A program has been created by NASA and NCI (National Cancer Institute) to exploit the possibility of carbon nanotube based biosensors to solve this problem for both's interest. In this talk, I will present our effort on the evaluation and novel design of carbon nanotubes as electrode biosensors with strategies to minimize background currents while maximizing signal intensity.The fabrication of nanotube electrode arrays, immobilization of molecular probes on nanotube electrodes and in vitro biosensor testing will also be discussed.

  7. Design and operation of a portable scanner for high performance microchip capillary array electrophoresis.

    Science.gov (United States)

    Scherer, James R; Liu, Peng; Mathies, Richard A

    2010-11-01

    We have developed a compact, laser-induced fluorescence detection scanner, the multichannel capillary array electrophoresis portable scanner (McCAEPs) as a platform for electrophoretic detection and control of high-throughput, integrated microfluidic devices for genetic and other analyses. The instrument contains a confocal optical system with a rotary objective for detecting four different fluorescence signals, a pneumatic system consisting of two pressure/vacuum pumps and 28 individual addressable solenoid valves for control of on-chip microvalves and micropumps, four Polymerase Chain Reaction (PCR) temperature control systems, and four high voltage power supplies for electrophoresis. The detection limit of the instrument is ~20 pM for on-chip capillary electrophoresis of fluorescein dyes. To demonstrate the system performance for forensic short tandem repeat (STR) analysis, two experiments were conducted: (i) electrophoretic separation and detection of STR samples on a 96-lane microfabricated capillary array electrophoresis microchip. Fully resolved PowerPlex(®) 16 STR profiles amplified from 1 ng of 9947A female standard DNA were successfully obtained; (ii) nine-plex STR amplification, sample injection, separation, and fluorescence detection of 100-copy 9948 male standard DNA in a single integrated PCR- capillary electrophoresis microchip. These results demonstrate that the McCAEPs can be used as a versatile control and detection instrument that operates integrated microfluidic devices for high-performance forensic human identification.

  8. Diagnostic accuracy of digital images for detection of artificial chemical proximal caries

    Energy Technology Data Exchange (ETDEWEB)

    Park, Geum Mee; Nah, Kyung Soo [Pusan National University College of Medicine, Busan (Korea, Republic of)

    2003-06-15

    To compare the diagnostic accuracy of proximal caries detection between Kodak Insight film and the Biomedisys CDX2000HQ digital (CCD) sensor. 156 proximal surfaces of extracted teeth, 78 of which had chemical artificial caries, were used in this study. Four observers interpreted the radiographs using a five-point confidence rating scale to record their diagnoses. The results were analyzed by receiver operating characteristic curves, ANOVA and Kappa values. Analysis using receiver operating characteristic curves revealed the areas under each curve which indicated a diagnostic accuracy of 0.951 in Insight and 0.952 in CDX2000HQ digital sensor. ANOVA revealed no significant differences between the two images with respect to caries detection. Kappa values indicated that the mean intra-observer agreement was 0.85 and inter-observer agreement 0.71 in conventional radiography. In digital radiography, the mean intra-observer agreement was 0.84 and inter-observer agreement 0.72. The results suggest that no significant difference exists between the two modalities for artificial caries detection and that CDX2000HQ was as good as Insight film for this purpose.

  9. Optimized acoustic biochip integrated with microfluidics for biomarkers detection in molecular diagnostics.

    Science.gov (United States)

    Papadakis, G; Friedt, J M; Eck, M; Rabus, D; Jobst, G; Gizeli, E

    2017-09-01

    The development of integrated platforms incorporating an acoustic device as the detection element requires addressing simultaneously several challenges of technological and scientific nature. The present work was focused on the design of a microfluidic module, which, combined with a dual or array type Love wave acoustic chip could be applied to biomedical applications and molecular diagnostics. Based on a systematic study we optimized the mechanics of the flow cell attachment and the sealing material so that fluidic interfacing/encapsulation would impose minimal losses to the acoustic wave. We have also investigated combinations of operating frequencies with waveguide materials and thicknesses for maximum sensitivity during the detection of protein and DNA biomarkers. Within our investigations neutravidin was used as a model protein biomarker and unpurified PCR amplified Salmonella DNA as the model genetic target. Our results clearly indicate the need for experimental verification of the optimum engineering and analytical parameters, in order to develop commercially viable systems for integrated analysis. The good reproducibility of the signal together with the ability of the array biochip to detect multiple samples hold promise for the future use of the integrated system in a Lab-on-a-Chip platform for application to molecular diagnostics.

  10. Newly established monoclonal antibody diagnostic assays for Schistosoma mansoni direct detection in areas of low endemicity.

    Directory of Open Access Journals (Sweden)

    Rafaella Fortini Queiroz Grenfell

    Full Text Available BACKGROUND: Current available methods for diagnosis of schistosomiasis mansoni lack sufficient sensitivity, which results in underreporting of infectious in areas of low endemicity. METHODOLOGY/PRINCIPAL FINDINGS: We developed three novel diagnostic methodologies for the direct detection of schistosome infection in serum samples. These three new methods were evaluated with positive patients from a low endemicity area in southeast Brazil. The basis of the assay was the production of monoclonal antibodies against the protein backbone of heavily glycosylated Circulating Cathodic Antigen (CCA. The antibodies were also selected for having no specificity to repeating poly-Lewis x units. Assays based on the detection CCA-protein should not encounter a limitation in sensitivity due to a biological background of this particular epitope. Three diagnostic methodologies were developed and validated, (i Immunomagnetic Separation based on improved incubation steps of non-diluted serum, (ii Direct Enzyme-linked Immunosorbent Assay and (iii Fluorescent Microscopy Analysis as a qualitative assay. The two quantitative assays presented high sensitivity (94% and 92%, respectively and specificity (100%, equivalent to the analysis of 3 stool samples and 16 slides by Kato-Katz, showing promising results on the determination of cure. CONCLUSIONS/SIGNIFICANCE: The Immunomagnetic Separation technique showed excellent correlation with parasite burden by Cohen coefficient. The qualitative method detected 47 positive individuals out of 50 with the analysis of 3 slides. This easy-to-do method was capable of discriminating positive from negative cases, even for patients with low parasite burden.

  11. Development of diagnostic SPR based biosensor for the detection of pharmaceutical compounds in saliva

    Science.gov (United States)

    Sonny, Susanna; Sesay, Adama M.; Virtanen, Vesa

    2010-11-01

    The aim of the study is to develop diagnostic tests for the detection of pharmaceutical compounds in saliva. Oral fluid is increasingly being considered as an ideal sample matrix. It can be collected non-invasively and causes less stress to the person being tested. The detection of pharmaceutical compounds and drugs in saliva can give valuable information on individual bases on dose response, usage, characterization and clinical diagnostics. Surface plasmon resonance (SPR) is a highly sensitive, fast and label free analytical technique for the detection of molecular interactions. The specific binding of measured analyte onto the active gold sensing surface of the SPR device induces a refractive index change that can be monitored. To monitor these pharmaceutical compounds in saliva the immunoassays were developed using a SPR instrument. The instrument is equipped with a 670nm laser diode and has two sensing channels. Monoclonal antibodies against the pharmaceutical compounds were used to specifically recognise and capture the compounds which intern will have an effect of the refractive index monitored. Preliminary results show that the immunoassays for cocaine and MDMA (3,4-methylenedioxymethamphetamine) are very sensitive and have linear ranges of 0.01 pg/ml - 1 ng/ml and 0.1 pg/ml - 100 ng/ml, respectively.

  12. Microchip-based ultrafast serodiagnostic assay for tuberculosis

    Science.gov (United States)

    Mani, Vigneshwaran; Paleja, Bhairav; Larbi, Karima; Kumar, Pavanish; Tay, Jo Ann; Siew, Jie Yee; Inci, Fatih; Wang, ShuQi; Chee, Cynthia; Wang, Yee Tang; Demirci, Utkan; De Libero, Gennaro; Singhal, Amit

    2016-01-01

    Access to point-of-care (POC), rapid, inexpensive, sensitive, and instrument-free tests for the diagnosis of tuberculosis (TB) remains a major challenge. Here, we report a simple and low-cost microchip-based TB ELISA (MTBE) platform for the detection of anti-mycobacterial IgG in plasma samples in less than 15 minutes. The MTBE employs a flow-less, magnet-actuated, bead-based ELISA for simultaneous detection of IgG responses against multiple mycobacterial antigens. Anti-trehalose 6,6′-dimycolate (TDM) IgG responses were the strongest predictor for differentiating active tuberculosis (ATB) from healthy controls (HC) and latent tuberculosis infections (LTBI). The TDM-based MTBE demonstrated superior sensitivity compared to sputum microscopy (72% vs. 56%) with 80% and 63% positivity among smear-positive and smear-negative confirmed ATB samples, respectively. Receiver operating characteristic analysis indicated good accuracy for differentiating ATB from HC (AUC = 0.77). Thus, TDM-based MTBE can be potentially used as a screening device for rapid diagnosis of active TB at the POC. PMID:27775039

  13. A compact 3D-printed interface for coupling open digital microchips with Venturi easy ambient sonic-spray ionization mass spectrometry.

    Science.gov (United States)

    Hu, Jie-Bi; Chen, Ting-Ru; Chang, Chia-Hsien; Cheng, Ji-Yen; Chen, Yu-Chie; Urban, Pawel L

    2015-03-07

    Digital microfluidics (DMF) based on the electrowetting-on-dielectric phenomenon is a convenient way of handling microlitre-volume aliquots of solutions prior to analysis. Although it was shown to be compatible with on-line mass spectrometric detection, due to numerous technical obstacles, the implementation of DMF in conjunction with MS is still beyond the reach of many analytical laboratories. Here we present a facile method for coupling open DMF microchips to mass spectrometers using Venturi easy ambient sonic-spray ionization operated at atmospheric pressure. The proposed interface comprises a 3D-printed body that can easily be "clipped" at the inlet of a standard mass spectrometer. The accessory features all the necessary connections for an open-architecture DMF microchip with T-shaped electrode arrangement, thermostatting of the microchip, purification of air (to prevent accidental contamination of the microchip), a Venturi pump, and two microfluidic pumps to facilitate transfer of samples and reagents onto the microchip. The system also incorporates a touch-screen panel and remote control for user-friendly operation. It is based on the use of popular open-source electronic modules, and can readily be assembled at low expense.

  14. Etching of glass microchips with supercritical water.

    Science.gov (United States)

    Karásek, Pavel; Grym, Jakub; Roth, Michal; Planeta, Josef; Foret, František

    2015-01-07

    A novel method of etching channels in glass microchips with the most tunable solvent, water, was tested as an alternative to common hydrogen fluoride-containing etchants. The etching properties of water strongly depend on temperature and pressure, especially in the vicinity of the water critical point. The chips were etched at the subcritical, supercritical and critical temperature of water, and the resulting channel shape, width, depth and surface morphology were studied by scanning electron microscopy and 3D laser profilometry. Channels etched with the hot water were compared with the chips etched with standard hydrogen fluoride-containing solution. Depending on the water pressure and temperature, the silicate dissolved from the glass could be re-deposited on the channel surface. This interesting phenomenon is described together with the conditions necessary for its utilization. The results illustrate the versatility of pure water as a glass etching and surface morphing agent.

  15. Scrambled eggs: A highly sensitive molecular diagnostic workflow for Fasciola species specific detection from faecal samples

    Science.gov (United States)

    Calvani, Nichola Eliza Davies; Windsor, Peter Andrew; Bush, Russell David

    2017-01-01

    Background Fasciolosis, due to Fasciola hepatica and Fasciola gigantica, is a re-emerging zoonotic parasitic disease of worldwide importance. Human and animal infections are commonly diagnosed by the traditional sedimentation and faecal egg-counting technique. However, this technique is time-consuming and prone to sensitivity errors when a large number of samples must be processed or if the operator lacks sufficient experience. Additionally, diagnosis can only be made once the 12-week pre-patent period has passed. Recently, a commercially available coprological antigen ELISA has enabled detection of F. hepatica prior to the completion of the pre-patent period, providing earlier diagnosis and increased throughput, although species differentiation is not possible in areas of parasite sympatry. Real-time PCR offers the combined benefits of highly sensitive species differentiation for medium to large sample sizes. However, no molecular diagnostic workflow currently exists for the identification of Fasciola spp. in faecal samples. Methodology/Principal findings A new molecular diagnostic workflow for the highly-sensitive detection and quantification of Fasciola spp. in faecal samples was developed. The technique involves sedimenting and pelleting the samples prior to DNA isolation in order to concentrate the eggs, followed by disruption by bead-beating in a benchtop homogeniser to ensure access to DNA. Although both the new molecular workflow and the traditional sedimentation technique were sensitive and specific, the new molecular workflow enabled faster sample throughput in medium to large epidemiological studies, and provided the additional benefit of speciation. Further, good correlation (R2 = 0.74–0.76) was observed between the real-time PCR values and the faecal egg count (FEC) using the new molecular workflow for all herds and sampling periods. Finally, no effect of storage in 70% ethanol was detected on sedimentation and DNA isolation outcomes; enabling

  16. Diagnostic value of minor salivary glands biopsy for the detection of Lewy pathology.

    Science.gov (United States)

    Folgoas, Emmanuelle; Lebouvier, Thibaud; Leclair-Visonneau, Laurène; Cersosimo, Maria-Graciela; Barthelaix, Annick; Derkinderen, Pascal; Letournel, Franck

    2013-09-13

    The recent demonstration of the presence of Lewy pathology in the submandibular glands of Parkinson's disease (PD) patients prompted us to evaluate the diagnostic performance of minor salivary gland biopsy for PD. Minor salivary glands were examined for Lewy pathology using phosphorylated alpha-synuclein antibody in 16 patients with clinically diagnosed PD and 11 control subjects with other neurological disorders. Abnormal accumulation of alpha-synuclein was found in 3 out of 16 PD patients. Two control subjects exhibited weak phosphorylated alpha-synuclein immunoreactivity. Our results do not support the use of minor salivary glands biopsy for the detection of Lewy pathology in living subjects.

  17. The diagnostic accuracy of carcinoembryonic antigen to detect colorectal cancer recurrence

    DEFF Research Database (Denmark)

    Sørensen, Caspar G; Karlsson, William K; Pommergaard, Hans-Christian;

    2016-01-01

    was to assess the diagnostic accuracy of CEA in detecting recurrence after intended curative surgery for primary colorectal cancer. METHODS: Systematic literature searches were performed in PubMed, EMBASE and Cochrane databases, and articles were chosen based on predefined inclusion criteria. Reference lists...... from included articles were manually searched for additional publications of relevance. RESULTS: Forty-two original studies with generally representative populations and long follow-up were included. Data were reported on outcomes from 9,834 CEA tests during follow-up. Reporting on the reference...

  18. Evaluation of Diagnostic Tests Using Information Theory for Multi-Class Diagnostic Problems and its Application for the Detection of Occlusal Caries Lesions

    Science.gov (United States)

    Arslan, Umut; Karaağaoğlu, Ergun; Özkan, Gökhan; Kanlı, Aydan

    2014-01-01

    Background: Several methods are available to evaluate the performance of the tests when the purpose of the diagnostic test is to discriminate between two possible disease states. However multi-class diagnostic problems frequently appear in many areas of medical science. Hence, there is a need for methods which will enable us to characterize the accuracy of diagnostic tests when there are more than two possible disease states. Aims: To show that two information theory measures, information content (IC) and proportional reduction in diagnostic uncertainty (PRDU), can be used for the evaluation of the performance of diagnostic tests for multi-class diagnostic problems that may appear in different areas of medical science. Study Design: Diagnostic accuracy study. Methods: Sixty freshly extracted permanent human molar and pre-molar teeth suspected to have occlusal caries lesions were selected for the study and were assessed by two experienced examiners. Each examiner performed two evaluations. Histological examination was used as the gold standard. The scores of the histological examination were defined as sound (n=11), enamel caries (n=22) and dentin caries (n=27). Diagnostic performance of i) visual inspection, ii) radiography, iii) laser fluorescence (LF) and iv) micro-computed tomography (M-CT) caries detection methods was evaluated by calculating IC and PRDU. Results: Micro-computed tomography examination was the best method among the diagnostic techniques for the diagnosis of occlusal caries in terms of both IC and PRDU. M-CT examination supplied the maximum diagnostic information about the diagnosis of occlusal caries in the first (IC: 1.056; p<0.05), (PRDU: 70.5%) and second evaluation (IC: 1.105; p<0.05), (PRDU: 73.8%) for the first examiner. M-CT examination was the best method among the diagnostic techniques for the second examiner in both the first (IC:1.105; p<0.05), (PRDU:73.8%) and second evaluation (IC:1.061; p<0.05), (PRDU:70.8%). IC and PRDU were

  19. Fungal disease detection in plants: Traditional assays, novel diagnostic techniques and biosensors.

    Science.gov (United States)

    Ray, Monalisa; Ray, Asit; Dash, Swagatika; Mishra, Abtar; Achary, K Gopinath; Nayak, Sanghamitra; Singh, Shikha

    2017-01-15

    Fungal diseases in commercially important plants results in a significant reduction in both quality and yield, often leading to the loss of an entire plant. In order to minimize the losses, it is essential to detect and identify the pathogens at an early stage. Early detection and accurate identification of pathogens can control the spread of infection. The present article provides a comprehensive overview of conventional methods, current trends and advances in fungal pathogen detection with an emphasis on biosensors. Traditional techniques are the "gold standard" in fungal detection which relies on symptoms, culture-based, morphological observation and biochemical identifications. In recent times, with the advancement of biotechnology, molecular and immunological approaches have revolutionized fungal disease detection. But the drawback lies in the fact that these methods require specific and expensive equipments. Thus, there is an urgent need for rapid, reliable, sensitive, cost effective and easy to use diagnostic methods for fungal pathogen detection. Biosensors would become a promising and attractive alternative, but they still have to be subjected to some modifications, improvements and proper validation for on-field use. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Diagnostic methods to cutaneous leishmaniasis detection in domestic dogs and cats*

    Science.gov (United States)

    Trevisan, Daliah Alves Coelho; Lonardoni, Maria Valdrinez Campana; Demarchi, Izabel Galhardo

    2015-01-01

    Cutaneous leishmaniasis is caused by different species of Leishmania. In domestic animals such as dogs and cats, the diagnostic consists of clinical, epidemiological and serological tests, which changes among countries all around the world. Because of this diversity in the methods selected, we propose this systematic literature review to identify the methods of laboratory diagnosis used to detect cutaneous leishmaniasis in domestic dogs and cats in the Americas. Articles published in the last 5 years were searched in PubMed, ISI Web of Science, LILACS and Scielo, and we selected 10 papers about cutaneous leishmaniasis in dogs and cats in the Americas. In Brazil, often the indirect immunofluorescence and enzyme immunoassay (ELISA) have been applied. Other countries like United States and Mexico have been using antigenic fractions for antibodies detections by Western blot. ELISA and Western blot showed a higher sensitivity and efficacy in the detection of leishmaniasis. Analysis of sensibility and specificity of the methods was rarely used. Although confirmatory to leishmaniasis, direct methods for parasites detection and polymerase chain reaction showed low positivity in disease detection. We suggested that more than one method should be used for the detection of feline and canine leishmaniasis. Serological methods such as Western blot and enzyme immunoassay have a high efficacy in the diagnosis of this disease. PMID:26734869

  1. Diagnostic methods to cutaneous leishmaniasis detection in domestic dogs and cats.

    Science.gov (United States)

    Trevisan, Daliah Alves Coelho; Lonardoni, Maria Valdrinez Campana; Demarchi, Izabel Galhardo

    2015-01-01

    Cutaneous leishmaniasis is caused by different species of Leishmania. In domestic animals such as dogs and cats, the diagnostic consists of clinical, epidemiological and serological tests, which changes among countries all around the world. Because of this diversity in the methods selected, we propose this systematic literature review to identify the methods of laboratory diagnosis used to detect cutaneous leishmaniasis in domestic dogs and cats in the Americas. Articles published in the last 5 years were searched in PubMed, ISI Web of Science, LILACS and Scielo, and we selected 10 papers about cutaneous leishmaniasis in dogs and cats in the Americas. In Brazil, often the indirect immunofluorescence and enzyme immunoassay (ELISA) have been applied. Other countries like United States and Mexico have been using antigenic fractions for antibodies detections by Western blot. ELISA and Western blot showed a higher sensitivity and efficacy in the detection of leishmaniasis. Analysis of sensibility and specificity of the methods was rarely used. Although confirmatory to leishmaniasis, direct methods for parasites detection and polymerase chain reaction showed low positivity in disease detection. We suggested that more than one method should be used for the detection of feline and canine leishmaniasis. Serological methods such as Western blot and enzyme immunoassay have a high efficacy in the diagnosis of this disease.

  2. Diagnostic Accuracy of Inverted and Unprocessed Digitized Periapical Radiographs for Detection of Peri-Implant Defects

    Directory of Open Access Journals (Sweden)

    Seyed Jalal Pourhashemi

    2016-04-01

    Full Text Available Objectives: This study aimed to compare the diagnostic accuracy of inverted and unprocessed digitized periapical radiographs for detection of peri-implant defects.Materials and Methods: A total of 30 osteotomy sites were prepared in three groups of control, study group 1 with 0.425 mm defects and study group 2 with 0.725 mm defects using the SIC and Astra Tech drill systems with 4.25mm and 4.85mm diameters. Small and large defects were randomly created in the coronal 8mm of 20 implant sites; implants (3.4mm diameter, 14.5mm length were then placed. Thirty periapical (PA radiographs were obtained using Digora imaging system (Soredex Corporation, Helsinki, Finland, size 2 photostimulable storage phosphor (PSP plate sensors (40.0mm×30.0mm and Scanora software. Unprocessed images were inverted using Scanora software by applying image inversion and a total of 60 images were obtained and randomly evaluated by four oral and maxillofacial radiologists. Data were analyzed using the t-test.Results: Significant differences were observed in absolute and complete sensitivity and specificity of the two imaging modalities for detection of small and large defects (P<0.05. Unprocessed digital images had a higher mean in terms of absolute sensitivity for detection of small defects, complete sensitivity for detection of large peri-implant defects and definite rule out of defects compared with inverted images.Conclusion: Unprocessed digital images have a higher diagnostic value for detection of small and large peri-implant defects and also for definite rule out of defects compared with inverted images.

  3. Rapid antigen detection test for respiratory syncytial virus diagnosis as a diagnostic tool,

    Directory of Open Access Journals (Sweden)

    Flávio da Silva Mesquita

    Full Text Available Abstract Objective: The aim of this study was to evaluate the QuickVue® RSV Test Kit (QUIDEL Corp, CA, USA as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. Methods: The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue® RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. Results: From 313 positive samples by immunofluorescence assays, 282 (90% were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue® RSV Test and viral load or specific strain. The QuickVue® RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. Conclusions: This study demonstrated that the QuickVue® RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics.

  4. Rapid antigen detection test for respiratory syncytial virus diagnosis as a diagnostic tool.

    Science.gov (United States)

    Mesquita, Flávio da Silva; Oliveira, Danielle Bruna Leal de; Crema, Daniela; Pinez, Célia Miranda Nunes; Colmanetti, Thaís Cristina; Thomazelli, Luciano Matsumia; Gilio, Alfredo Elias; Vieira, Sandra Elisabeth; Martinez, Marina Baquerizo; Botosso, Viviane Fongaro; Durigon, Edison Luiz

    The aim of this study was to evaluate the QuickVue(®) RSV Test Kit (QUIDEL Corp, CA, USA) as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue(®) RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. From 313 positive samples by immunofluorescence assays, 282 (90%) were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue(®) RSV Test and viral load or specific strain. The QuickVue(®) RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. This study demonstrated that the QuickVue(®) RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics. Copyright © 2016 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.

  5. Retrograde-viewing device improves adenoma detection rate in colonoscopies for surveillance and diagnostic workup

    Institute of Scientific and Technical Information of China (English)

    Peter D Siersema; Amit Rastogi; Anke M Leufkens; Paul A Akerman; Kassem Azzouzi; Richard I Rothstein; Frank P Vleggaar

    2012-01-01

    AIM:To determine which patients might benefit most from retrograde viewing during colonoscopy through subset analysis of randomized,controlled trial data.METHODS:The Third Eye(R) Retroscope(R) Randomized Clinical Evaluation (TERRACE) was a randomized,controlled,multicenter trial designed to evaluate the efficacy of a retrograde-viewing auxiliary imaging device that is used during colonoscopy to provide a second video image which allows viewing of areas on the proximal aspect of haustral folds and flexures that are difficult to see with the colonoscope's forward view.We performed a post-hoc analysis of the TER-RACE data to determine whether certain subsets of the patient population would gain more benefit than others from use of the device.Subjects were patients scheduled for colonoscopy for screening,surveillance or diagnostic workup,and each underwent same-day tandem examinations with standard colonoscopy (SC)and Third Eye colonoscopy (TEC),randomized to SC followed by TEC or vice versa.RESULTS:Indication for colonoscopy was screening in 176/345 subjects (51.0%),surveillance after previous polypectomy in 87 (25.2%) and diagnostic workup in 82 (23.8%).In 4 subjects no indication was specified.Previously reported overall results had shown a net additional adenoma detection rate (ADR) with TEC of 23.2% compared to SC.Relative risk (RR) of missing adenomas with SC vs TEC as the initial procedure was 1.92 (P =0.029).Post-hoc subset analysis shows additional ADRs for TEC compared to SC were 4.4% for screening,35.7% for surveillance,55.4% for diagnostic and 40.7% for surveillance and diagnostic combined.The RR of missing adenomas with SC vs TEC was 1.11 (P =0.815) for screening,3.15 (P =0.014) for surveillance,8.64 (P =0.039) for diagnostic and 3.34(P =0.003) for surveillance and diagnostic combined.Although a multivariate Poisson regression suggested gender as a possibly significant factor,subset analysis showed that the difference between genders was

  6. Diagnostic Values of Laser Fluorescence Device with Other Techniques in Occlusal Caries Detection

    Directory of Open Access Journals (Sweden)

    A. Chehresaz

    2009-06-01

    Full Text Available Objective: The aim of the present study was to compare the performance of laser fluorescence (LF method with other conventional diagnostic techniques in detection of smallocclusal caries in permanent teeth.Materials and Methods: Prior to this in vitro diagnostic study, a pilot study assessed intra-examiner reliability and reproducibility. The occlusal surfaces of 90 extracted human premolars were examined with four diagnostic methods: probing, visual inspection, bitewing(BW radiographs, and LF. The teeth were then sectioned for the purpose of histological examination. The data were analyzed using SPSS 15 software, and sensitivity,specificity and other diagnostic criteria of the techniques were calculated.Results: The intra-examiner reproducibility for probing and also radiographic techniques was 100%. The corresponded figure for LF (88% was more than visual inspection (82%.The highest level of validity of the examiner turned out to be in probing technique(76.9%. Sensitivity of visual inspection, probing, and LF methods was 54.5% and that of BW radiography was 27.5%. Specificity and efficiency of LF method were 84.8% and 81.1%, respectively. Probing and visual inspection showed the highest specificity (97.5% and 94.9, respectively and efficiency (92.2% and 90%, respectively among the methods.Conclusion: Specificity and efficiency of LF method were lower compared to those of other methods. Among all the investigated methods, the most efficient methods in the diagnosis of small occlusal caries in permanent teeth were probing and visual inspection,respectively.

  7. Microchip electrophoresis with background electrolyte containing polyacrylic acid and high content organic solvent in cyclic olefin copolymer microchips for easily adsorbed dyes.

    Science.gov (United States)

    Wei, Xuan; Sun, Ping; Yang, Shenghong; Zhao, Lei; Wu, Jing; Li, Fengyun; Pu, Qiaosheng

    2016-07-29

    Plastic microchips can significantly reduce the fabrication cost but the adsorption of some analytes limits their application. In this work, background electrolyte containing ionic polymer and high content of organic solvent was adopted to eliminate the analyte adsorption and achieve highly efficient separation in microchip electrophoresis. Two dyes, rhodamine 6G (Rh6G) and rhodamine B (RhB) were used as the model analytes. By using methanol as the organic solvent and polyacrylic acid (PAA) as a multifunctional additive, successful separation of the two dyes within 75μm id. microchannels was realized. The role of PAA is multiple, including viscosity regulator, selectivity modifier and active additive for counteracting analyte adsorption on the microchannel surface. The number of theoretical plate of 7.0×10(5)/m was attained within an effective separation distance of 2cm using background electrolyte consisting 80% methanol, 0.36% PAA and 30mmol/L phosphate at pH 5.0. Under optimized conditions, relative standard deviations of Rh6G and RhB detection (n=5) were no more than 1.5% for migration time and 2.0% for peak area, respectively. The limit of detection (S/N=3) was 0.1nmol/L for Rh6G. The proposed technique was applied in the determination of both Rh6G and RhB in chilli powder and lipstick samples with satisfactory recoveries of 81.3-103.7%.

  8. A Diagnostic Algorithm for the Detection of Clostridium difficile-Associated Diarrhea.

    Science.gov (United States)

    Yoldaş, Özlem; Altındiş, Mustafa; Cufalı, Davut; Aşık, Gülşah; Keşli, Recep

    2016-01-01

    Clostridium difficile is a common cause of hospital-acquired diarrhea, which is usually associated with previous antibiotic use. The clinical manifestations of C. difficile infection (CDI) may range from mild diarrhea to fulminant colitis. Clostridium difficile should be considered in diarrhea cases with a history of antibiotic use within the last 8 weeks (community-associated CDI) or with a hospital stay of at least 3 days, regardless of the duration of antibiotic use (hospital-acquired CDI). This study investigated the frequency of CDI in diarrheic patients and evaluated the efficacy of the triple diagnostic algorithm that is proposed here for C. difficile detection. Cross-sectional study. In this study, we compared three methods currently employed for C. difficile detection using 95 patient stool samples: an enzyme immunoassay (EIA) for toxin A/B (C. diff Toxin A+B; Diagnostic Automation Inc.; Calabasas, CA, USA), an EIA for glutamate dehydrogenase (GDH) (C. DIFF CHEK-60TM, TechLab Inc.; Blacksburg, VA, USA), and a polymerase chain reaction (PCR)-based assay (GeneXpert(®) C. difficile; Cepheid, Sunnyvale, CA, USA) that detects C. difficile toxin genes and conventional methods as well. In this study, 50.5% of the patients were male, 50 patients were outpatients, 32 were from inpatient clinics and 13 patients were from the intensive care unit. Of the 95 stool samples tested for GDH, 28 were positive. Six samples were positive by PCR, while nine samples were positive for toxin A/B. The hypervirulent strain NAP-1 and binary toxin was not detected. The rate of occurrence of toxigenic C. difficile was 5.1% in the samples. Cefaclor, ampicillin-sulbactam, ertapenem, and piperacillin-tazobactam were the most commonly used antibiotics by patients preceding the onset of diarrhea. Among the patients who were hospitalized in an intensive care unit for more than 7 days, 83.3% were positive for CDI by PCR screening. If the PCR test is accepted as the reference: C. difficile

  9. A Robust Automated Cataract Detection Algorithm Using Diagnostic Opinion Based Parameter Thresholding for Telemedicine Application

    Directory of Open Access Journals (Sweden)

    Shashwat Pathak

    2016-09-01

    Full Text Available This paper proposes and evaluates an algorithm to automatically detect the cataracts from color images in adult human subjects. Currently, methods available for cataract detection are based on the use of either fundus camera or Digital Single-Lens Reflex (DSLR camera; both are very expensive. The main motive behind this work is to develop an inexpensive, robust and convenient algorithm which in conjugation with suitable devices will be able to diagnose the presence of cataract from the true color images of an eye. An algorithm is proposed for cataract screening based on texture features: uniformity, intensity and standard deviation. These features are first computed and mapped with diagnostic opinion by the eye expert to define the basic threshold of screening system and later tested on real subjects in an eye clinic. Finally, a tele-ophthamology model using our proposed system has been suggested, which confirms the telemedicine application of the proposed system.

  10. An in vivo comparison of two diagnostic methods in secondary caries detection.

    Directory of Open Access Journals (Sweden)

    Kazem Saber Hamishaki

    2014-02-01

    Full Text Available The aim of this study was to compare the level of agreement between four operators with different levels of experience for two methods of detecting secondary enamel and dentin carious lesions in composite restored teeth.Sixty teeth of 40 patients with with secondary carious lesions in the composite resin were selected. The teeth were examined by two methods; visual inspection and a laser fluorescence device (DIAGNO dent pen 2190 by four operators including an undergraduate student, a dentist with 5 years of clinical experience, a general dentist with 12 years of clinical experience and an oral and maxillofacial radiologist. Cohen's kappa statistic was applied in order to assess the agreement between the diagnoses performed by the four operators with each diagnostic method.The diagnosis performed by different operators achieved an excellent agreement with high ICC.DIAGNOdent can be a useful device for secondary caries detection in posterior teeth as an adjunct to visual examination.

  11. Diagnostic value of meat juice in early detection of classical swine fever infection

    DEFF Research Database (Denmark)

    Lohse, Louise; Uttenthal, Åse; Rasmussen, Thomas Bruun

    2011-01-01

    To evaluate the diagnostic potential of meat juice for early detection of Classical swine fever virus (CSFV), meat juice and serum samples from pigs experimentally infected with different strains of CSFV were compared for virus load. From all samples, viral RNA was extracted by automated procedure...... before real-time reverse transcription polymerase chain reaction analysis was performed. Viral RNA was detected in meat juice, but at a lower level than in corresponding serum. Sensitivity was calculated to 91% and specificity to 97%. Disagreements between meat juice and serum results were found when...... samples originated from pigs infected with low virulence CSFV strains and/or when samples were collected within the first days after infection. In conclusion, while not the first choice for sample material for CSFV diagnosis, meat juice may constitute a useful alternative for herd-based studies or when...

  12. An In Vivo Comparison of Two Diagnostic Methods in Secondary Caries Detection

    Science.gov (United States)

    Hamishaki, Kazem Saber; Chiniforush, Nasim; Monzavi, Abbas; Khazarazifard, Mohammad Javad

    2014-01-01

    Objective: The aim of this study was to compare the level of agreement between four operators with different levels of experience for two methods of detecting secondary enamel and dentin carious lesions in composite restored teeth. Materials and Methods: Sixty teeth of 40 patients with with secondary carious lesions in the composite resin were selected. The teeth were examined by two methods; visual inspection and a laser fluorescence device (DIAGNO dent pen 2190) by four operators including an undergraduate student, a dentist with 5 years of clinical experience, a general dentist with 12 years of clinical experience and an oral and maxillofacial radiologist. Cohen's kappa statistic was applied in order to assess the agreement between the diagnoses performed by the four operators with each diagnostic method. Results: The diagnosis performed by different operators achieved an excellent agreement with high ICC. Conclusion: DIAGNOdent can be a useful device for secondary caries detection in posterior teeth as an adjunct to visual examination. PMID:24910672

  13. Microbial Diagnostic Microarrays for the Detection and Typing of Food- and Water-Borne (Bacterial) Pathogens.

    Science.gov (United States)

    Kostić, Tanja; Sessitsch, Angela

    2011-10-14

    Reliable and sensitive pathogen detection in clinical and environmental (including food and water) samples is of greatest importance for public health. Standard microbiological methods have several limitations and improved alternatives are needed. Most important requirements for reliable analysis include: (i) specificity; (ii) sensitivity; (iii) multiplexing potential; (iv) robustness; (v) speed; (vi) automation potential; and (vii) low cost. Microarray technology can, through its very nature, fulfill many of these requirements directly and the remaining challenges have been tackled. In this review, we attempt to compare performance characteristics of the microbial diagnostic microarrays developed for the detection and typing of food and water pathogens, and discuss limitations, points still to be addressed and issues specific for the analysis of food, water and environmental samples.

  14. Microbial Diagnostic Microarrays for the Detection and Typing of Food- and Water-Borne (Bacterial Pathogens

    Directory of Open Access Journals (Sweden)

    Tanja Kostić

    2011-10-01

    Full Text Available Reliable and sensitive pathogen detection in clinical and environmental (including food and water samples is of greatest importance for public health. Standard microbiological methods have several limitations and improved alternatives are needed. Most important requirements for reliable analysis include: (i specificity; (ii sensitivity; (iii multiplexing potential; (iv robustness; (v speed; (vi automation potential; and (vii low cost. Microarray technology can, through its very nature, fulfill many of these requirements directly and the remaining challenges have been tackled. In this review, we attempt to compare performance characteristics of the microbial diagnostic microarrays developed for the detection and typing of food and water pathogens, and discuss limitations, points still to be addressed and issues specific for the analysis of food, water and environmental samples.

  15. Evaluation of the diagnostic value of 64 simultaneously measured autoantibodies for early detection of gastric cancer

    Science.gov (United States)

    Werner, Simone; Chen, Hongda; Butt, Julia; Michel, Angelika; Knebel, Phillip; Holleczek, Bernd; Zörnig, Inka; Eichmüller, Stefan B.; Jäger, Dirk; Pawlita, Michael; Waterboer, Tim; Brenner, Hermann

    2016-01-01

    Autoantibodies against tumor-associated antigens (TAAs) have been suggested as biomarkers for early detection of gastric cancer. However, studies that systematically assess the diagnostic performance of a large number of autoantibodies are rare. Here, we used bead-based multiplex serology to simultaneously measure autoantibody responses against 64 candidate TAAs in serum samples from 329 gastric cancer patients, 321 healthy controls and 124 participants with other diseases of the upper digestive tract. At 98% specificity, sensitivities for the 64 tested autoantibodies ranged from 0–12% in the training set and a combination of autoantibodies against five TAAs (MAGEA4 + CTAG1 + TP53 + ERBB2_C + SDCCAG8) was able to detect 32% of the gastric cancer patients at a specificity of 87% in the validation set. Sensitivities for early and late stage gastric cancers were similar, while chronic atrophic gastritis, a precursor lesion of gastric cancer, was not detectable. However, the 5-marker combination also detected 26% of the esophageal cancer patients. In conclusion, the tested autoantibodies and combinations alone did not reach sufficient sensitivity for gastric cancer screening. Nevertheless, some autoantibodies, such as anti-MAGEA4, anti-CTAG1 or anti-TP53 and their combinations could possibly contribute to the development of cancer early detection tests (not necessarily restricted to gastric cancer) when being combined with other markers. PMID:27140836

  16. Bacillus anthracis diagnostic detection and rapid antibiotic susceptibility determination using 'bioluminescent' reporter phage.

    Science.gov (United States)

    Schofield, David A; Sharp, Natasha J; Vandamm, Joshua; Molineux, Ian J; Spreng, Krista A; Rajanna, Chythanya; Westwater, Caroline; Stewart, George C

    2013-11-01

    Genetically modified phages have the potential to detect pathogenic bacteria from clinical, environmental, or food-related sources. Herein we assess an engineered 'bioluminescent' reporter phage (Wß::luxAB) as a clinical diagnostic tool for Bacillus anthracis, the etiological agent of anthrax. Wß::luxAB is able to rapidly (within minutes) detect a panel of B. anthracis strains by transducing a bioluminescent phenotype. The reporter phage displays species specificity by its inability, or significantly reduced ability, to detect members of the closely related Bacillus cereus group and other common bacterial pathogens. Using spiked clinical specimens, Wß::luxAB detects B. anthracis within 5 h at clinically relevant concentrations, and provides antibiotic susceptibility information that mirrors the CLSI method, except that data are obtained at least 5-fold faster. Although anthrax is a treatable disease, a positive patient prognosis is dependent on timely diagnosis and appropriate therapy. Wß::luxAB rapidly detects B. anthracis and determines antibiotic efficacy, properties that will help patient outcome.

  17. Automatic detection of paroxysmal atrial fibrillation in patients with ischaemic stroke: better than routine diagnostic workup?

    Science.gov (United States)

    Uphaus, T; Grings, A; Gröschel, S; Müller, A; Weber-Krüger, M; Wachter, R; Gröschel, K

    2017-07-01

    Prolonged electrocardiogram (ECG) monitoring after ischaemic stroke increases the diagnostic yield of paroxysmal atrial fibrillation (pAF). In order to facilitate the additional workload involved in ECG analysis due to prolonged monitoring times, we investigated the effectiveness of pAF detection with an automated software algorithm (SA) in comparison to the routine staff-based analysis (RA) during standard stroke-unit care. Therefore, patients with acute ischaemic stroke or transitory ischaemic attack presenting with sinus rhythmus on the admission ECG and no history of atrial fibrillation were prospectively included. A 24-h Holter ECG assessment was performed using either RA based on a computer-aided evaluation and subsequent review by a cardiologist or a commercially available automated SA. In the case of discordant results concerning the occurrence of pAF between the two methods, the data underwent an independent external rating. Of 809 prospectively enrolled patients, 580 patients fulfilled the inclusion criteria. pAF was ultimately diagnosed in 3.3% of the cohort (19 patients). SA and RA correctly diagnosed pAF in 17 patients resulting in a comparable diagnostic effectiveness of the analysis methods (sensitivity: SA 89.5% vs. RA 89.5%; specificity: SA 99.3% vs. RA 99.1%; κ, 0.686; P Holter ECG reaches a high diagnostic effectiveness for the detection of pAF and can be used for a rapid and resource-saving analysis of ECG data to deal with prolonged monitoring times. © 2017 The Authors. European Journal of Neurology published by John Wiley & Sons Ltd on behalf of European Academy of Neurology.

  18. Detection of hemorrhage source: the diagnostic value of post-mortem CT-angiography.

    Science.gov (United States)

    Palmiere, C; Binaghi, S; Doenz, F; Bize, P; Chevallier, C; Mangin, P; Grabherr, S

    2012-10-10

    The aim of this study was to compare the diagnostic value of post-mortem computed tomography angiography (PMCTA) to conventional, ante-mortem computed tomography (CT)-scan, CT-angiography (CTA) and digital subtraction angiography (DSA) in the detection and localization of the source of bleeding in cases of acute hemorrhage with fatal outcomes. The medical records and imaging scans of nine individuals who underwent a conventional, ante-mortem CT-scan, CTA or DSA and later died in the hospital as a result of an acute hemorrhage were reviewed. Post-mortem computed tomography angiography, using multi-phase post-mortem CTA, as well as medico-legal autopsies were performed. Localization accuracy of the bleeding was assessed by comparing the diagnostic findings of the different techniques. The results revealed that data from ante-mortem and post-mortem radiological examinations were similar, though the PMCTA showed a higher sensitivity for detecting the hemorrhage source than did ante-mortem radiological investigations. By comparing the results of PMCTA and conventional autopsy, much higher sensitivity was noted in PMCTA in identifying the source of the bleeding. In fact, the vessels involved were identified in eight out of nine cases using PMCTA and only in three cases through conventional autopsy. Our study showed that PMCTA, similar to clinical radiological investigations, is able to precisely identify lesions of arterial and/or venous vessels and thus determine the source of bleeding in cases of acute hemorrhages with fatal outcomes.

  19. Diagnostic evaluation of RT-PCR-ELISA for the detection of rabies virus.

    Science.gov (United States)

    Aravindhbabu, R P; Manoharan, S; Ramadass, P

    2014-01-01

    Rabies is primarily a disease of terrestrial and airborne mammals. In most cases, rabies is diagnosed primarily on the basis of clinical symptoms and signs, and a corroborative history of or evidence of an animal bite, death of an animal and incomplete or no vaccination following exposure. The facility for laboratory diagnosis and confirmation of rabies is available in only a few institutions in India. Diagnostic tests using conventional assays like fluorescent antibody test (FAT) are unreliable at times, despite the clinical diagnosis. Currently, there are a number of molecular tests that can be used to complement conventional tests in rabies diagnosis. We have developed and evaluated an RT-PCR-ELISA using a panel of brain tissue samples from rabies suspected animals of various species. This assay was able to detect rabies virus genome in all the 43 samples that were previously tested positive for rabies. Moreover this assay was shown to be 100 % sensitive and specific in detecting the rabies virus genome in post-mortem brain tissue samples from different species of animals. Our pilot study shows the potential of this assay as an alternative diagnostic test when the samples are unsuitable for use in FAT and also a supplementary test to FAT. In addition, the region of nucleoprotein gene amplified using this assay can be used for the molecular investigation of geographical origin of the field strains.

  20. Multicenter Evaluation of Clinical Diagnostic Methods for Detection and Isolation of Campylobacter spp. from Stool.

    Science.gov (United States)

    Fitzgerald, Collette; Patrick, Mary; Gonzalez, Anthony; Akin, Joshua; Polage, Christopher R; Wymore, Kate; Gillim-Ross, Laura; Xavier, Karen; Sadlowski, Jennifer; Monahan, Jan; Hurd, Sharon; Dahlberg, Suzanne; Jerris, Robert; Watson, Renee; Santovenia, Monica; Mitchell, David; Harrison, Cassandra; Tobin-D'Angelo, Melissa; DeMartino, Mary; Pentella, Michael; Razeq, Jafar; Leonard, Celere; Jung, Carrianne; Achong-Bowe, Ria; Evans, Yaaqobah; Jain, Damini; Juni, Billie; Leano, Fe; Robinson, Trisha; Smith, Kirk; Gittelman, Rachel M; Garrigan, Charles; Nachamkin, Irving

    2016-05-01

    The use of culture-independent diagnostic tests (CIDTs), such as stool antigen tests, as standalone tests for the detection of Campylobacter in stool is increasing. We conducted a prospective, multicenter study to evaluate the performance of stool antigen CIDTs compared to culture and PCR for Campylobacter detection. Between July and October 2010, we tested 2,767 stool specimens from patients with gastrointestinal illness with the following methods: four types of Campylobacter selective media, four commercial stool antigen assays, and a commercial PCR assay. Illnesses from which specimens were positive by one or more culture media or at least one CIDT and PCR were designated "cases." A total of 95 specimens (3.4%) met the case definition. The stool antigen CIDTs ranged from 79.6% to 87.6% in sensitivity, 95.9 to 99.5% in specificity, and 41.3 to 84.3% in positive predictive value. Culture alone detected 80/89 (89.9% sensitivity) Campylobacter jejuni/Campylobacter coli-positive cases. Of the 209 noncases that were positive by at least one CIDT, only one (0.48%) was positive by all four stool antigen tests, and 73% were positive by just one stool antigen test. The questionable relevance of unconfirmed positive stool antigen CIDT results was supported by the finding that noncases were less likely than cases to have gastrointestinal symptoms. Thus, while the tests were convenient to use, the sensitivity, specificity, and positive predictive value of Campylobacter stool antigen tests were highly variable. Given the relatively low incidence of Campylobacter disease and the generally poor diagnostic test characteristics, this study calls into question the use of commercially available stool antigen CIDTs as standalone tests for direct detection of Campylobacter in stool. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  1. Problems Associated with the Microchip Data of Stray Dogs and Cats Entering RSPCA Queensland Shelters

    Science.gov (United States)

    Lancaster, Emily; Rand, Jacquie; Collecott, Sheila; Paterson, Mandy

    2015-01-01

    Simple Summary Microchip identification has become an important tool to reunite stray dogs and cats with their owners, and is now compulsory in most states of Australia. Improvement of the microchipping system in Australia is limited by a lack of published Australian data documenting the problems experienced by shelter staff when using microchip data to contact the owner of a stray animal. In this study we determine the character and frequency of inaccurate microchip data to identify weaknesses in the current microchipping system. This information could be used to develop strategies that increase the accuracy of microchip data that will increase the reclaiming of stray animals. Abstract A lack of published information documenting problems with the microchip data for the reclaiming of stray animals entering Australian shelters limits improvement of the current microchipping system. A retrospective study analysing admission data for stray, adult dogs (n = 7258) and cats (n = 6950) entering the Royal Society for the Prevention of Cruelty to Animals (RSPCA) Queensland between January 2012 and December 2013 was undertaken to determine the character and frequency of microchip data problems and their impact on outcome for the animal. Only 28% of dogs and 9% of cats were microchipped, and a substantial proportion (37%) had problems with their data, including being registered to a previous owner or organisation (47%), all phone numbers incorrect/disconnected (29%), and the microchip not registered (14%). A higher proportion of owners could be contacted when the microchip had no problems, compared to those with problems (dogs, 93% vs. 70%; cats, 75% vs. 41%). The proportion of animals reclaimed declined significantly between microchipped animals with no data problems, microchipped animals with data problems and non-microchipped animals—87%, 69%, and 37%, respectively, for dogs and 61%, 33%, and 5%, respectively, for cats. Strategies are needed to increase the accuracy of

  2. Zeolite nanoparticle modified microchip reactor for efficient protein digestion.

    Science.gov (United States)

    Huang, Yi; Shan, Wei; Liu, Baohong; Liu, Yun; Zhang, Yahong; Zhao, Yue; Lu, Haojie; Tang, Yi; Yang, Pengyuan

    2006-04-01

    An enzymatic microreactor has been fabricated based on the poly(methyl methacrylate) (PMMA) microchchip surface-modified with zeolite nanoparticles. By introducing the silanol functional groups, the surface of PMMA microchannel has been successfully modified with silicalite-1 nanoparticle for the first time due to its large external surface area and high dispersibility in solutions. Trypsin can be stably immobilized in the microchannel to form a bioreactor using silica sol-gel matrix. The immobilization of enzyme can be realized with a stable gel network through a silicon-oxygen-silicon bridge via tethering to those silanol groups, which has been investigated by scanning electron microscopy and microchip capillary electrophoresis with laser-induced fluorescence detection. The maximum proteolytic rate constant of the immobilized trypsin is measured to be about 6.6 mM s(-1). Using matrix assisted laser desorption and ionization time-of-flight mass spectrometry, the proposed microreactor provides an efficient digestion of cytochrome c and bovine serum albumin at a fast flow rate of 4.0 microL min(-1), which affords a very short reaction time of less than 5 s.

  3. Diagnostic Performance of Gray-scale Sonographic Findings for the Detection of Acute Pyelonephritis

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hui Jin; Kim, Young Jun; Park, Hee Sun; Jeon, Hae Jeong; Park, Hyoung Keun; Paick, Sung Hyun; Kim, Hyeong Gon; Lho, Yong Soo; Jung, Sung Il [Konkuk University School of Medicine, Seoul (Korea, Republic of)

    2012-03-15

    To evaluate the diagnostic performance of gray-scale renal sonographic findings for the diagnosis of acute pyelonephritis (APN) by using computed tomography as a reference standard. We retrospectively reviewed gray-scale renal sonographic findings of 48 patients for the detection of APN. All patients had clinical symptoms such as fever, flank pain, or dysuria and were confirmed as APN by contrast- enhanced CT. The presence of sonographic findings such as renal swelling, alteration of the parenchymal echogenicity, wall thickening of the renal pelvis, loss of the renal sinus fat echogenicity, and loss of the corticomedullary differentiation were evaluated. We also categorized all patients into mild APN or severe APN groups according to the volume of the morbid renal parenchyma on contrast-enhanced CT, and evaluated the aforementioned sonographic findings between the two groups. Overall diagnostic sensitivity, specificity, and accuracy of gray-scale renal ultrasonography (US) for the detection of APN were 32.5%, 72.0%, and 58.5%, respectively. The sensitivity and specificity of each sonographic finding were measured for each group. Renal swelling sensitivity and specificity were 33.8% and 70.8% for the mild APN group, but 45.8% and 66.7% for the severe APN group. Sensitivity and specificity for alteration of the parenchymal echogenicity were 41.7% and 79.2% for the mild APN group, but 58.3% and 66.7% for the severe APN group. The sensitivity and specificity for wall thickening of the renal pelvis was 37.5% and 95.8% for the mild APN group, but 50.0% and 95.8% for the severe APN group. The sensitivity and specificity of loss of the renal sinus fat echogenicity were 12.5% and 83.3% for the mild APN group, but 12.5% and 91.7% for the severe APN group. The sensitivity and specificity of the loss of the corticomedullary differentiation were 12.5% and 95.8% for the mild APN group, but 20.8% and 75.0% for the severe APN group. There was no significant difference of gray

  4. Potential diagnostic value of serum p53 antibody for detecting esophageal cancer: a meta-analysis.

    Directory of Open Access Journals (Sweden)

    Jun Zhang

    Full Text Available BACKGROUND: Mutant p53 protein overexpression has been reported to induce serum antibodies against p53. Various studies assessing the diagnostic value of serum p53 antibody in patients with esophageal cancer remain controversial. This study aims to comprehensively and quantitatively summarize the potential diagnostic value of serum p53 antibody in esophageal cancer. METHODS: We systematically searched PubMed and Embase until 31st May 2012, without language restriction. Studies were assessed for quality using QUADAS (quality assessment of studies of diagnostic accuracy. Positive likelihood ratio (PLR and negative likelihood ratio (NLR were pooled separately and compared with overall accuracy measures diagnostic odds ratio (DOR and symmetric summary receiver operating characteristic (sROC. The PLR and NLR and their 95% confidence interval (CI were calculated using a fixed effects model according to the Mantel-Haensed method and random effects model based on the work of Der Simonian and laird, respectively. RESULTS: Fifteen studies (cases = 1079, controls = 2260 met the inclusion criteria for the meta-analysis. Approximately 53.33% (8/15 of the included studies were of high quality (QUADAS score≥8, which were retrospective case-control studies. The summary estimates for quantitative analysis of serum p53 antibody in the diagnosis of esophageal cancer were PLR 6.95 (95% CI: 4.77-9.51, NLR 0.75 (95%CI: 0.72-0.78 and DOR 9.65 (95%CI: 7.04-13.22. However, we found significant heterogeneity between NLRs. CONCLUSIONS: The current evidence suggests serum p53 antibody has a potential diagnostic value for esophageal cancer. However, its discrimination power is not perfect because of low sensitivity. IMPACT: These results suggest that s-p53-antibody may be useful for monitoring residual tumor cells and for aiding in the selection of candidates for less invasive treatment procedures because of the high specificity of s-p53-antibody. Further studies

  5. Pulse laser head with monolithic thermally bonded microchip operating at 1.5 μm wavelength

    Science.gov (United States)

    Młyńczak, Jarosław; Kopczyński, Krzysztof; Belghachem, Nabil; Kisielewski, Jarosław; Stepień, Ryszard; Wychowaniec, Marek; Galas, Jacek; Litwin, Dariusz; CzyŻewski, Adam

    2016-12-01

    On the basis of thermally bonded Er,Yb:glass/Co:MALO microchip a laser head pumped by fiber coupled laser diode was designed. The performance of the laser head were investigated and the main output parameters were determined. The energy over 40 μJ in 3.8 ns pulse with repetition rate of 0.735 kHz was achieved. The laser head characterized by such parameters can successfully be used in tele-detection applications.

  6. Novel peptidoglycan-based diagnostic devices for detection of wound infection.

    Science.gov (United States)

    Hasmann, Andrea; Wehrschuetz-Sigl, Eva; Kanzler, Gertraud; Gewessler, Ulrike; Hulla, Elisabeth; Schneider, Konstantin P; Binder, Barbara; Schintler, Michael; Guebitz, Georg M

    2011-09-01

    Detection of wound infection is based on evaluation of the well-known signs of inflammation like rubor (redness), calor (heat), tumor (swelling), and dolor (pain) by medical doctors and/or time-consuming procedures requiring special machinery. There is currently no rapid diagnostic device available for the indication of wound infection, which would especially be helpful in home care of chronic ulcer patients. In this study, a new concept for a fast diagnostic tool for wound infection based on lysozyme and elastase triggered release of dye from a peptidoglycan matrix was investigated. The matrix consisted of alginate/agarose and peptidoglycan covalently labeled with Remazol brilliant blue. Lysozyme activity in postoperative wounds and decubitus wound fluids was significantly elevated upon infection (4830 ± 1848 U mL(-1)) compared to noninfected wounds (376 ± 240 U mL(-1)). Consequently, incubation of 8% (w/v) labeled agarose/peptidoglycan blend layers with infected wound fluid samples for 2 h at 37 °C resulted in a 4-fold higher amount of dye released than measured for noninfected wounds. For alginate/peptidoglycan beads, a 7-fold higher amount of dye was released in case of infected wound fluid samples compared to noninfected ones. Apart from lysozyme, proteases [i.e., gelatinase matrix metalloproteinase MMP-2 and MMP-9 and elastase] were detected in wound fluids (e.g., using Western blotting). When dosed in ratios typical for wounds, a slight synergistic effect was measured for peptidoglycan hydrolysis (i.e., dye release) between lysozyme and these proteases. Incubation of a double-layer system consisting of stained and nonstained peptidoglycan with infected wound fluids resulted in a color change from yellow to blue, thus allowing simple visual detection of wound infection. Copyright © 2011 Elsevier Inc. All rights reserved.

  7. Ultrasensitive Nanoelectrospray Ionization-Mass Spectrometry using Poly(dimethylsiloxane) Microchips with Monolithically Integrated Emitters

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Xuefei; Kelly, Ryan T.; Tang, Keqi; Smith, Richard D.

    2010-09-01

    Poly(dimethylsiloxane) (PDMS) is the most widely used substrate for microfluidic devices as it enables facile fabrication and has other distinctive properties. However, for applications involving highly sensitive nanoelectrospray ionization mass spectrometry (nanoESI-MS) detection, the use of PDMS microdevices has been hindered by the leaching of uncross-linked oligomers and other contaminants from the substrate that yields a large background of chemical noise in the mass spectra. A more general challenge is that microfluidic devices containing integrated electrospray emitters are frequently unable to operate stably in the nanoflow regime where the best sensitivity is achieved. In this report, we extracted the contaminants from PDMS substrates using a series of solvents, eliminating the background observed when untreated PDMS microchips are used for nanoESI-MS. Optimization of the integrated emitter geometry enabled stable operation at flow rates as low as 10 nL/min. Peptide concentrations of 1 nM were readily detected, representing ~170 zmol of consumed analyte, and an extrapolated detection limit of ~40 zmol; these are the lowest mass and concentration detection limits reported to date for a microchip having an integrated electrospray emitter.

  8. A Study on the Diagnostic Detection Ability of the Artificial Proximal Caries by Digora

    Energy Technology Data Exchange (ETDEWEB)

    Oh, Kyung Ran; Choi, Eui Hwan; Kim, Jae Duck [Dept. of Oral and Maxillofacial Radiology, College of Dentistry, Chosun University, Kwangju (Korea, Republic of)

    1998-08-15

    Digora system is an intraoral indirect digital radiography system utilizing storage phosphor image plate. It has wide dynamic range which allows it to decrease the patient's exposure time and may increase diagnostic ability through image processing (such as edge enhancement, grey scale conversion, brightness change, and contrast enhancement). And also, it can transmit and storage image information. The purpose of this study was to evaluate the diagnostic ability of artificial proximal caries between Conventional radiograph and Digora images (unenhanced image, brightness and contrast controlled image, and edge enhanced image). ROC (Receiver Operating Characteristic) analysis, paired t-tests, and F-tests were done for the statistical evaluation of detectability. The following results were acquired: 1. In Grade I lesions, the mean ROC areas of Conventional radiograph, Digora unenhanced image, Digora controlled image, and Digora edge enhanced image were 0.953, 0.933, 0.965, 0.978 (p>0.05). 2. In Grade II lesions, the mean ROC areas of Conventional radiograph, Digora unenhanced image, Digora controlled image, and Digora edge enhanced image were 0.969, 0.964, 0.988, 0.994. Among theses areas, there was just statistical significance between Diagnostic abilities of Digora edge enhanced image and Conventional radiograph (p<0.05). 3. In the Interobserver variability, the ROC curve areas of Digora edge enhanced image was lowerest in these areas, regardless of the Carious lesion depths. In conclusion, intraoral indirect digital system, Digora system, has the potential possibility as an alternative of Conventional radiograph in the diagnosis of proximal caries.

  9. [The value of diagnostic ultrasound for detecting occult inguinal hernia in patients with groin pain].

    Science.gov (United States)

    Lorenzini, Cesare; Sofia, Lorenzo; Pergolizzi, Francesca Pia; Trovato, Massimo

    2008-01-01

    The value of diagnostic ultrasound for detecting occult inguinal hernia in The aim of this study was to demonstrate the accuracy of ultrasound in diagnosing occult hernia in patients with recurrent inguinal pain and a negative clinical examination. Over a period of three years, a total of 51 patients were referred for ultrasound examination with clinically suspected occult inguinal hernia. Patients with positive US scans were offered surgery, while those with negative US findings were offered further imaging or other diagnostic tests. The ultrasound examination of the inguinal region was conducted using a 5-10 MHz linear probe. The scan was performed with the patient in the supine and erect positions, in a relaxed state, as well as during coughing and during a Valsalva manoeuvre. Overall, ultrasound diagnosed 20 inguinal hernias and all 20 patients with positive scans underwent surgery. Surgery confirmed the ultrasound diagnosis in 19 patients, only 1/20 having no hernia at operation (100% ultrasound sensitivity and 96.9% specificity). Patients undergoing surgery showed complete symptom resolution at a three-month follow-up. This study confirms that ultrasound is capable of accurately diagnosing groin hernia and this may justify its use in patients with chronic groin pain due to a suspected occult hernia.

  10. Establishing a diagnostic system for detecting Ralstonia solanacearum and genetic differentiation using RAPD molecular markers

    Directory of Open Access Journals (Sweden)

    Edisson Chavarro Mesa

    2007-02-01

    Full Text Available A polymerase chain reaction-based diagnostic test (PCR has been developed for amplifying a región and obtaining a 292 bp product by using specific 16S rDNA primers for the rapid and precise identification of the causative agent (Ralstonia solanacearum of bacterial withering of potato in asymptomatic tubers. The bacteria was isolated from potato tubers and banana fruit using culturing techniques and immunological and molecular ELISA-NCM and PCR tests, respectively. PCR detected the presence of R. solanacearum on asymptomatic tubers by contrast with ELISA-NCM which did not detect this pathogen. Analysing random amplified polymorphic DNA (RAPD led to differentiating and grouping R. solanacearum by geographical región and bacterial strain, suggesting that differences exist amongst existing collections according to their place of origin, presenting high genetic variability. The results showed that PCR is a sensitive and specific test for detecting R. solanacearum and can therefore be implemented as a method for controlling this pathogen in seed production and certification programmes in áreas free of the disease. The pathogen has been shown to be genetically heterogeneous according to the samples' geographical área thereby hampering control in áreas of Colombia experiencing phytosanitary problems with R. solanacearum in potato crops Key words: bacterial withered, moko, PCR-16S rADN, ELISA-NCM, PCR-RAPD.

  11. Performance of newer and conventional diagnostic methods in detection of drug sensitive and resistant tuberculous meningitis

    Directory of Open Access Journals (Sweden)

    Sangamithra Neelakantan

    2014-09-01

    Full Text Available Objective: To compare the performance of traditional and newer diagnostic methods for diagnosis of tuberculous meningitis (TBM along with detection of drug resistant TBM. Methods: Single cerebrospinal fluids sample from 281 suspected TBM patients was processed during August 2011 to July 2012 for acid fast bacilli (AFB examination by Ziehl-Neelsen and auramine staining methods, AFB culture on Lowenstein-Jensen culture media, by microscope observation of drug susceptibility testing assay, and nucleic acid amplification tests by inhouse conventional PCR and in-house real time PCR targeting IS6110 insertion sequence. All the isolates were subjected to drug susceptibility testing for isoniazid, rifampicin, ethambutol and streptomycin by 1% proportion method. Results: The percentage positivity by AFB smear examination and culture on Lowenstein-Jensen media was 21% (59/281, 11.4% (32/281 respectively. The detection rate by conventional PCR was 33.1% (93/281. The real time PCR showed positivity rate of 44.8% (126/281. The extra detection by real time PCR was 11.7%. Only one isolate was multiple drug resistant and 22 (68.8% were pan-susceptible. Remaining eight isolates showed either mono/poly drug resistant to first line antitubercular drugs. Conclusions: Real time PCR is a more sensitive and rapid method if it is appropriately adopted in clinical practice for diagnosis of TBM. Prevalence of multiple drug resistance Mycobacterium tuberculosis causing TBM is low.

  12. Laser diagnostic technology for early detection of pathogen infestation in orange fruits

    Energy Technology Data Exchange (ETDEWEB)

    Giubileo, Gianfranco, E-mail: gianfranco.giubileo@frascati.enea.i [ENEA Frascati, Via E. Fermi 45, 00044 (Italy); Lai, Antonella; Piccinelli, Delinda [ENEA Frascati, Via E. Fermi 45, 00044 (Italy); Puiu, Adriana [Tor Vergata University of Rome, Faculty of Engineering, Via del Politecnico 1, 00133 Rome (Italy)

    2010-11-11

    Due to an increased expectation of food products that respect high quality and safety standards, there is a need for the growth of accurate, fast, objective and non-destructive technologies for quality determination of food and agricultural products. For this purpose, a diagnostic system based on laser photoacoustic spectroscopy (LPAS) was developed at ENEA Frascati Molecular Spectroscopy Laboratory (Italy). In the design of the photoacoustic detector, particular emphasis was placed in attaining a high sensitivity in detecting ethylene (ET) down to sub-parts per billion level (minimum detectable concentration 0.2 ppb). This was required due to the necessity to monitor and follow up ET production at a single fruit scale. ET is normally synthesised in very low amounts by healthy citrus fruits; however stress conditions such as pathogen attack may induce a substantial increase in the synthesised ET. In the present paper, the comparison between the ET emitted by healthy oranges (Citrus sinensis L. Osbeck) cv Navel and by Phytophthora citrophthora infested Navel orange fruits are reported. The obtained results show a well evident increase in ET emission from the infested fruit with respect to the healthy one, even 24 h after the inoculation with the pathogen; at that time the tissue necrosis was not yet visible, and the fruit was also not yet damaged. The possibility to perform a real time non-destructive detection of ET traces makes the LPAS a powerful tool for monitoring the healthy state of the citrus fruits.

  13. Molecular method for the detection of Andes hantavirus infection: validation for clinical diagnostics.

    Science.gov (United States)

    Vial, Cecilia; Martinez-Valdebenito, Constanza; Rios, Susana; Martinez, Jessica; Vial, Pablo A; Ferres, Marcela; Rivera, Juan C; Perez, Ruth; Valdivieso, Francisca

    2016-01-01

    Hantavirus cardiopulmonary syndrome is a severe disease caused by exposure to New World hantaviruses. Early diagnosis is difficult due to the lack of specific initial symptoms. Antihantavirus antibodies are usually negative until late in the febrile prodrome or the beginning of cardiopulmonary phase, while Andes hantavirus (ANDV) RNA genome can be detected before symptoms onset. We analyzed the effectiveness of quantitative reverse transcription polymerase chain reaction (RT-qPCR) as a diagnostic tool detecting ANDV-Sout genome in peripheral blood cells from 78 confirmed hantavirus patients and 166 negative controls. Our results indicate that RT-qPCR had a low detection limit (~10 copies), with a specificity of 100% and a sensitivity of 94.9%. This suggests the potential for establishing RT-qPCR as the assay of choice for early diagnosis, promoting early effective care of patients, and improving other important aspects of ANDV infection management, such as compliance of biosafety recommendations for health personnel in order to avoid nosocomial transmission.

  14. [Plasma cholesterol determination in birds--a diagnostic tool for detection of organophosphate and carbamate intoxication].

    Science.gov (United States)

    Kiesau, B; Kummerfeld, N

    1998-07-01

    An investigation was done on the clinical usefulness of the dry chemistry analyzer Vitros DT 60 II for determination of avian plasma cholinesterase. The analytical reliability of the method, evaluated by precision and accuracy, proved to be high for plasma of numerous pet and wild birds. Values of normal plasma-cholinesterase activity were established for different psittacine and European wild birds. Significant differences in physiologic plasma-cholinesterase activity were noted between closely related species as well as between juvenile and adult birds. These findings emphasize the necessity to use control values of the same species and age group for comparison. Dry chemistry plasma-cholinesterase determination can be used as a diagnostic tool for detection of organophosphate and carbamate poisonings in the majority of investigated birds.

  15. Multi-wavelength diagnostic properties of Galactic Planetary Nebulae detected by GLIMPSE-I

    CERN Document Server

    Cohen, Martin; Green, Anne J; Miszalski, Brent; Frew, David J; Murphy, Tara

    2010-01-01

    We uniformly analyze 136 optically detected PNe and candidates from the GLIMPSE-I survey in order to to develop robust, multi-wavelength, classification criteria to augment existing diagnostics and provide pure PN samples. PNe represent powerful astrophysical probes. They are important dynamical tracers, key sources of ISM chemical enrichment, windows into late stellar evolution, and potent cosmological yardsticks. But their utility depends on separating them unequivocally from the many nebular mimics which can strongly resemble bona fide PNe in traditional optical images and spectra. We merge new PNe from the carefully evaluated, homogeneous MASH-I and MASH-II surveys, which offer a wider evolutionary range of PNe than hitherto available, with previously known PNe classified by SIMBAD. Mid-infrared (MIR) measurements vitally complement optical data because they reveal other physical processes and morphologies via fine-structure lines, molecular bands and dust. MIR colour-colour planes, optical emission line ...

  16. New detection system and signal processing for the tokamak ISTTOK heavy ion beam diagnostic

    Energy Technology Data Exchange (ETDEWEB)

    Henriques, R. B.; Nedzelskiy, I. S.; Malaquias, A.; Fernandes, H. [Associacao Euratom/IST, Instituto de Plasmas e Fusao Nuclear, Instituto Superior Tecnico, Universidade Tecnica de Lisboa, 1049-001 Lisboa (Portugal)

    2012-10-15

    The tokamak ISTTOK heavy ion beam diagnostic (HIBD) operates with a multiple cell array detector (MCAD) that allows for the plasma density and the plasma density fluctuations measurements simultaneously at different sampling volumes across the plasma. To improve the capability of the plasma density fluctuations investigations, a new detection system and new signal conditioning amplifier have been designed and tested. The improvements in MCAD design are presented which allow for nearly complete suppression of the spurious plasma background signal by applying a biasing potential onto special electrodes incorporated into MCAD. The new low cost and small size transimpedance amplifiers are described with the parameters of 400 kHz, 10{sup 7} V/A, 0.4 nA of RMS noise, adequate for the plasma density fluctuations measurements.

  17. A diagnostic PCR assay for the detection of an Australian epidemic strain of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Murphy Anna

    2010-07-01

    Full Text Available Abstract Background Chronic lung infection with the bacterium Pseudomonas aeruginosa is one of the hallmarks of cystic fibrosis (CF and is associated with worsening lung function, increased hospitalisation and reduced life expectancy. A virulent clonal strain of P. aeruginosa (Australian epidemic strain I; AES-I has been found to be widespread in CF patients in eastern Australia. Methods Suppression subtractive hybridization (SSH was employed to identify genetic sequences that are present in the AES-I strain but absent from the sequenced reference strain PAO1. We used PCR to evaluate the distribution of several of the AES-I loci amongst a collection of 188 P. aeruginosa isolates which was comprised of 35 AES-I isolates (as determined by PFGE, 78 non-AES-I CF isolates including other epidemic CF strains as well as 69 P. aeruginosa isolates from other clinical and environmental sources. Results We have identified a unique AES-I genetic locus that is present in all 35 AES-I isolates tested and not present in any of the other 153 P. aeruginosa strains examined. We have used this unique AES-I locus to develop a diagnostic PCR and a real-time PCR assay to detect the presence of P. aeruginosa and AES-I in patient sputum samples. Conclusions We have developed diagnostic PCR assays that are 100% sensitive and 100% specific for the P. aeruginosa strain AES-I. We have also shown that Whatman FTA® Elute cards may be used with PCR-based assays to rapidly detect the presence of P. aeruginosa strains in CF sputum.

  18. A new diagnostic score to detect osteoporosis in patients undergoing lumbar spine MRI

    Energy Technology Data Exchange (ETDEWEB)

    Bandirali, Michele; Messina, Carmelo [Universita degli Studi di Milano, Scuola di Specializzazione in Radiodiagnostica, Milano (Italy); Di Leo, Giovanni; Papini, Giacomo Davide Edoardo [IRCCS Policlinico San Donato, Radiology Unit, San Donato Milanese (Italy); Sconfienza, Luca Maria; Sardanelli, Francesco [IRCCS Policlinico San Donato, Radiology Unit, San Donato Milanese (Italy); Universita degli Studi di Milano, Dipartimento di Scienze Biomediche per la Salute, San Donato Milanese (Italy); Ulivieri, Fabio Massimo [IRCCS Fondazione Ca' Granda Ospedale Maggiore Policlinico, Mineralometria Ossea Computerizzata e Ambulatorio Malattie Metabolismo Minerale e Osseo, Servizio di Medicina Nucleare, Milano (Italy)

    2015-10-15

    Signal intensity of lumbar-spine at magnetic resonance imaging (MRI) correlates to bone mineral density (BMD). Our aim was to define a quantitative MRI-based score to detect osteoporosis on lumbar-spine MRI. After Ethics Committee approval, we selected female patients who underwent both lumbar-spine MRI and dual-energy X-ray absorptiometry (DXA) and a reference group of 131 healthy females (20-29 years) who underwent lumbar-spine MRI. We measured the intra-vertebral signal-to-noise ratio in L1-L4. We introduced an MRI-based score (M-score), on the model of T-score. M-score diagnostic performance in diagnosing osteoporosis was estimated against DXA using receiver operator characteristic (ROC) analysis. We included 226 patients (median age 65 years), 70 (31 %) being osteoporotic at DXA. MRI signal-to-noise ratio correlated to BMD (r = -0.677, P < 0.001). M-score negatively correlated to T-score (r = -0.682, P < 0.001). Setting a 90 %-specificity, an M-score threshold of 5.5 was found, distinguishing osteoporosis from non-osteoporosis (sensitivity 54 %; ROC AUC 0.844). Thirty-one (14 %) patients had a fragility fracture, with osteoporosis detected in 15 (48 %) according to M-score and eight (26 %) according to T-score (P = 0.016). M-score obtained on lumbar spine MRI is a quantitative method correlating with osteoporosis. Its diagnostic value remains to be demonstrated on a large prospective cohort of patients. (orig.)

  19. Development and evaluation of virtual refrigerant mass flow sensors for fault detection and diagnostics

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Woohyun; Braun, J.

    2016-03-05

    Refrigerant mass flow rate is an important measurement for monitoring equipment performance and enabling fault detection and diagnostics. However, a traditional mass flow meter is expensive to purchase and install. A virtual refrigerant mass flow sensor (VRMF) uses a mathematical model to estimate flow rate using low-cost measurements and can potentially be implemented at low cost. This study evaluates three VRMFs for estimating refrigerant mass flow rate. The first model uses a compressor map that relates refrigerant flow rate to measurements of inlet and outlet pressure, and inlet temperature measurements. The second model uses an energy-balance method on the compressor that uses a compressor map for power consumption, which is relatively independent of compressor faults that influence mass flow rate. The third model is developed using an empirical correlation for an electronic expansion valve (EEV) based on an orifice equation. The three VRMFs are shown to work well in estimating refrigerant mass flow rate for various systems under fault-free conditions with less than 5% RMS error. Each of the three mass flow rate estimates can be utilized to diagnose and track the following faults: 1) loss of compressor performance, 2) fouled condenser or evaporator filter, 3) faulty expansion device, respectively. For example, a compressor refrigerant flow map model only provides an accurate estimation when the compressor operates normally. When a compressor is not delivering the expected flow due to a leaky suction or discharge valve or other internal fault, the energy-balance or EEV model can provide accurate flow estimates. In this paper, the flow differences provide an indication of loss of compressor performance and can be used for fault detection and diagnostics.

  20. Serial dilution microchip for cytotoxicity test

    Science.gov (United States)

    Bang, Hyunwoo; Lim, Sun Hee; Lee, Young Kyung; Chung, Seok; Chung, Chanil; Han, Dong-Chul; Chang, Jun Keun

    2004-08-01

    Today's pharmaceutical industry is facing challenges resulting from the vast increases in sample numbers produced by high-throughput screening (HTS). In addition, the bottlenecks created by increased demand for cytotoxicity testing (required to assess compound safety) are becoming a serious problem. We have developed a polymer PDMS (polydimethylsiloxane) based microfluidic device that can perform a cytotoxicity test in a rapid and reproducible manner. The concept that the device includes is well adjustable to automated robots in huge HTS systems, so we can think of it as a potential dilution and delivery module. Cytotoxicity testing is all about the dilution and dispensing of a drug sample. Previously, we made a PDMS based microfluidic device which automatically and precisely diluted drugs with a buffer solution with serially increasing concentrations. This time, the serially diluted drug solution was directly delivered to 96 well plates for cytotoxicity testing. Cytotoxic paclitaxel solution with 2% RPMI 1640 has been used while carrying out cancerous cell based cytotoxicity tests. We believe that this rapid and robust use of the PDMS microchip will overcome the growing problem in cytotoxicity testing for HTS.

  1. Rapid bonding of Pyrex glass microchips.

    Science.gov (United States)

    Akiyama, Yoshitake; Morishima, Keisuke; Kogi, Atsuna; Kikutani, Yoshikuni; Tokeshi, Manabu; Kitamori, Takehiko

    2007-03-01

    A newly developed vacuum hot press system has been specially designed for the thermal bonding of glass substrates in the fabrication process of Pyrex glass microchemical chips. This system includes a vacuum chamber equipped with a high-pressure piston cylinder and carbon plate heaters. A temperature of up to 900 degrees C and a force of as much as 9800 N could be applied to the substrates in a vacuum atmosphere. The Pyrex substrates bonded with this system under different temperatures, pressures, and heating times were evaluated by tensile strength tests, by measurements of thickness, and by observations of the cross-sectional shapes of the microchannels. The optimal bonding conditions of the Pyrex glass substrates were 570 degrees C for 10 min under 4.7 N/mm(2) of applied pressure. Whereas more than 16 h is required for thermal bonding with a conventional furnace, the new system could complete the whole bonding processes within just 79 min, including heating and cooling periods. Such improvements should considerably enhance the production rate of Pyrex glass microchemical chips. Whereas flat and dust-free surfaces are required for conventional thermal bonding, especially without long and repeated heating periods, our hot press system could press a fine dust into glass substrates so that even the areas around the dust were bonded. Using this capability, we were able to successfully integrate Pt/Ti thin film electrodes into a Pyrex glass microchip.

  2. A microchip platform for structural oncology applications

    Science.gov (United States)

    Winton, Carly E; Gilmore, Brian L; Demmert, Andrew C; Karageorge, Vasilea; Sheng, Zhi; Kelly, Deborah F

    2016-01-01

    Recent advances in the development of functional materials offer new tools to dissect human health and disease mechanisms. The use of tunable surfaces is especially appealing as substrates can be tailored to fit applications involving specific cell types or tissues. Here we use tunable materials to facilitate the three-dimensional (3D) analysis of BRCA1 gene regulatory complexes derived from human cancer cells. We employed a recently developed microchip platform to isolate BRCA1 protein assemblies natively formed in breast cancer cells with and without BRCA1 mutations. The captured assemblies proved amenable to cryo-electron microscopy (EM) imaging and downstream computational analysis. Resulting 3D structures reveal the manner in which wild-type BRCA1 engages the RNA polymerase II (RNAP II) core complex that contained K63-linked ubiquitin moieties—a putative signal for DNA repair. Importantly, we also determined that molecular assemblies harboring the BRCA15382insC mutation exhibited altered protein interactions and ubiquitination patterns compared to wild-type complexes. Overall, our analyses proved optimal for developing new structural oncology applications involving patient-derived cancer cells, while expanding our knowledge of BRCA1’s role in gene regulatory events. PMID:27583302

  3. Cryogenic Tm:YAP microchip laser

    Science.gov (United States)

    Hubka, Zbyněk.; Å ulc, Jan; Jelínková, Helena; Nejezchleb, Karel; Å koda, Václav

    2016-04-01

    The spectral characteristics of laser active media, and thus those of the laser output, are temperature dependent. Specifically, in almost every crystal host, cooling to low temperatures leads to better heat removal, a higher efficiency and output power, and a reduced lasing threshold. Tm-ion doped lasers have an emission wavelength around 2 μm and are important in medicine for soft tissue cutting and hemostasis, as well as in LIDAR or atmosphere sensing technology. This paper presents the performance-temperature dependency of a 4 at. % doped Tm:YAP microchip. During the experiment the Tm:YAP crystal was placed inside an evacuated liquid nitrogen cryostat on a cooling finger. As its temperature was varied from 80 K to 340 K, changes were observed in the absorption spectrum, ranging from 750 nm to 2000 nm and in the fluorescence spectrum from 1600 nm to 2050 nm. Fluorescence lifetime was seen to rise and fall with decreasing temperature. The laser was pumped by a 792 nm laser diode and at 80 K the maximum output peak power of the laser was 4.6 W with 23 % slope efficiency and 0.6 W threshold, compared to 2.4 W output peak power, 13 % slope efficiency and 3.3 W threshold when at 340 K. The laser emission wavelength changed from 1883 nm to 1993 nm for 80 K and 300 K, respectively.

  4. Organically modified sols as pseudostationary phases for microchip electrophoresis.

    Science.gov (United States)

    Pumera, Martin; Wang, Joseph; Grushka, Eli; Lev, Ovadia

    2007-04-30

    We demonstrate that the selectivity of microchip electrophoresis separations is greatly improved by the presence of organically modified silica (Ormosil) sols in the run buffer. A negatively-charged N-(trimethoxysilylpropyl)ethylenediamine triacetic-acid (TETT)-based sol is used for improving the selectivity between nitroaromatic explosives and a methyltrimethoxysilane (MTMOS)-based sol is employed for enhancing the microchip separation of environmental pollutants, aminophenols. These sols are added to the run buffer and act as pseudostationary phases. Their presence in the run buffer changes the apparent mobility of studied solutes, and leads to a higher resolution. The observed mobilities changes reflect the interactions between the Ormosil sols and the solutes. Relevant experimental variables have been characterized and optimized. The diverse chemistry of Ormosil sols should be extremely useful for tailoring the selectivity of a wide range of electrophoresis microchip separations.

  5. A disposable laser print-cut-laminate polyester microchip for multiplexed PCR via infra-red-mediated thermal control.

    Science.gov (United States)

    Ouyang, Yiwen; Duarte, Gabriela R M; Poe, Brian L; Riehl, Paul S; dos Santos, Fernando M; Martin-Didonet, Claudia C G; Carrilho, Emanuel; Landers, James P

    2015-12-11

    Infrared (IR)-mediated thermal cycling system, a method proven to be a effective for sub-μL scale polymerase chain reaction (PCR) on microchips, has been integrated with DNA extraction and separation on a glass microchip in a fully integrated micro Total Analysis System by Easley et al., in 2006. IR-PCR has been demonstrated on both glass and PMMA microdevices where the fabrication (bonding) is not trivial. Polyester-toner (PeT) microfluidic devices have significant potential as cost-effective, disposable microdevices as a result of the ease of fabrication (∼$0.25 USD and <10 min per device) and availability of commercial substrates. For the first time, we demonstrate here the thermal cycling in PeT microchips on the IR-PCR system. Undesirable IR absorption by the black-toner bonding layer was eliminated with a spatial filter in the form of an aluminum foil mask. The solution heating rate for a black PeT microchip using a tungsten lamp was 10.1 ± 0.7 °C s(-1) with a cooling rate of roughly -12 ± 0.9 °C s(-1) assisted by forced air cooling. Dynamic surface passivation strategies allowed the successful amplification of a 520 bp fragment of the λ-phage genome (in 11 min) and a 1500 bp region of Azospirillum brasilense. Using a centrosymmetric chamber configuration in a multichamber PeT microchip, homogenous temperature distribution over all chambers was achieved with inter-chamber temperature differences at annealing, extension and denaturing steps of less than ±2 °C. The effectiveness of the multichamber system was demonstrated with the simultaneous amplification of a 390 bp amplicon of human β-globin gene in five PeT PCR microchambers. The relative PCR amplification efficiency with a human β-globin DNA fragment ranged from 70% to 90%, in comparison to conventional thermal cyclers, with an inter-chamber standard deviation of ∼10%. Development of PeT microchips for IR-PCR has the potential to provide rapid, low-volume amplification while

  6. Diagnostic performance of MRI for detection of intestinal fistulas in patients with complicated inflammatory bowel conditions

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt, S.; Meuwly, J.Y.; Meuli, R.; Schnyder, P.; Denys, A. [Universitaire Vaudois - CHUV, Service de radiodiagnostic et radiologie interventionnelle, Centre Hospitalier, Lausanne (Switzerland); Chevallier, P. [Hopital Archet II, Imagerie Medicale, Nice (France); Bessoud, B. [Hopital Kremlin-Bicetre, Radiologie Generale, Kremlin-Bicetre (France); Felley, C. [University Hospital, CHUV, Service de Gastroenterologie, Lausanne (Switzerland)

    2007-11-15

    The diagnostic performance of magnetic resonance imaging (MRI) for detection of intestinal fistulas, other than perianal, in patients with known complicated inflammatory bowel conditions (CIBC) was investigated. Our study group consisted of 20 patients (12 women, mean age 43 years) with CIBC, including Crohn's disease (n=13), colonic diverticulitis (n=3), colitis after radiotherapy (n=3) and of postoperative origin (n=1). Eleven surgically proven enteral fistulas were known in ten (50%) of these patients, being of enterovesical (n=3), enterocolic (n=2), enteroenteral (n=2), rectovaginal (n=2), rectovaginovesical (n=1) and of entercutaneous (n=1) localisation. The other ten patients (50%), used as the control group, showed MR features of CIBC, although without any fistulous tract. Multiplanar T1- and T2-weighted sequences had been performed, including gadolinium-enhanced acquisition with fat saturation (1.5 T). MR findings were independently blindly and retrospectively reviewed by three radiologists for the presence and etiology of any fistula, as well as visualization and characterization of the fistulous tract. Results were compared with surgical findings (n=16) and clinical evolution (n=4). Interobserver agreement was calculated. Interobserver agreement kappa for fistula detection was 0.71. Overall sensitivity, specificity and accuracy for fistula detection were 78.6%, 75% and 77.2%, respectively. Sensitivity for fistula characterization was 80.6%, with visualization of the fistulous tract in all cases, whereby T1-weighted gadolinium-enhanced fat-saturated images were considered the most useful sequences. Gadolinium-enhanced MRI is a reliable and reproducible tool for detection of enteral fistulas secondary to inflammatory conditions. (orig.)

  7. Detection of ingested cocaine-filled packets-Diagnostic value of unenhanced CT

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt, Sabine [Department of Radiology, University Hospital-CHUV, Rue du Bugnon, 1011 Lausanne (Switzerland)], E-mail: sabine.schmidt@chuv.ch; Hugli, Olivier [Medical Emergency Department, University Hospital-CHUV, Rue du Bugnon, 1011 Lausanne (Switzerland); Rizzo, Elena; Lepori, Domenico; Gudinchet, F. [Department of Radiology, University Hospital-CHUV, Rue du Bugnon, 1011 Lausanne (Switzerland); Yersin, Bertrand [Medical Emergency Department, University Hospital-CHUV, Rue du Bugnon, 1011 Lausanne (Switzerland); Schnyder, Pierre; Meuwly, Jean-Yves [Department of Radiology, University Hospital-CHUV, Rue du Bugnon, 1011 Lausanne (Switzerland)

    2008-07-15

    Purpose: Emergency departments are facing nowadays an increasing number of illegal drug-related health problems, associated with medicolegal and/or social consequences. Body stuffers are street cocaine dealers, who either store wrapped packets of drugs in their rectum or hastily swallow them, prompted by fear of police's arrest. These packets can be life threatening in case of leakage. We evaluate the diagnostic value of unenhanced multidetector CT (MDCT) for detection of cocaine-filled packets (CFP) ingested by body stuffers in a phantom model. Materials and methods: Our phantom simulated normal bowel contents in which a varying number of true and false CFP were randomly mixed. Both only differ in radiological density. During 18 different reading sessions, four radiologists independently evaluated the presence and number of true and false CFP. Interobserver agreement, sensitivity, specificity, positive and negative predictive value were calculated. Results: Interobserver agreement for detection of any packets, for visualization of true, and false CFP was good ({kappa} = 0.63, 0.74 and 0.58, respectively). Sensitivity, specificity, positive and negative predictive value for detection of any packets was 95.6%, 100%, 100% and 62.5%, respectively; for visualization of the true CFP 86.5%, 100%, 100% and 77.6%, respectively; and for the false packets 98.1%, 65%, 88.6% and 87.5%, respectively. Conclusion: Unenhanced MDCT without bowel preparation is a fast, reliable and easily reproducible imaging modality for the immediate detection of ingested CFP, thus facilitating medicolegal management of body stuffers.

  8. High repetition rate passively Q-switched fiber and microchip lasers for optical resolution photoacoustic imaging

    Science.gov (United States)

    Shi, Wei; Utkin, Ilya; Ranasinghesagara, Janaka; Pan, Lei; Godwal, Yogesh; Kerr, Shaun; Zemp, Roger J.; Fedosejevs, Robert

    2010-02-01

    Optical-resolution photoacoustic microscopy is a novel imaging technology for visualizing optically-absorbing superficial structures in vivo with lateral spatial resolution determined by optical focusing rather than acoustic detection. Since scanning of the illumination spot is required, the imaging speed is limited by the scanning speed and the laser pulse repetition rate. Unfortunately, lasers with high-repetition rate and suitable pulse durations and energies are difficult to find. We are developing compact laser sources for this application. Passively Q-switched fiber and microchip lasers with pulse repetition rates up to 300 kHz are demonstrated. Using a diode-pumped microchip laser fiber-coupled to a large mode-area Yb-doped fiber amplifier we obtained 60μJ 1-ns pulses at the frequency-doubled 532-nm wavelength. The pulse-repetition rate was determined by the power of the microchip laser pump source at 808nm and may exceed 10 kHz. Additionally, a passively Q-switched fiber laser utilizing a Yb-doped double-cladding fiber and an external saturable absorber has shown to produce 250ns pulses at repetition rates of 100-300 KHz. A photoacoustic probe enabling flexible scanning of the focused output of these lasers consisted of a 45-degree glass prism in an optical index-matching fluid. Photoacoustic signals exiting the sample are deflected by the prism to an ultrasound transducer. Phantom studies with a 7.5-micron carbon fiber demonstrate the ability to image with optical rather than acoustic resolution. We believe that the high pulse-repetition rates and the potentially compact and fiber-coupled nature of these lasers will prove important for clinical imaging applications where realtime imaging performance is essential.

  9. Diagnostic accuracy of 16-slice multidetector-row CT for detection of in-stent restenosis vs detection of stenosis in nonstented coronary arteries.

    Science.gov (United States)

    Kefer, Joelle M; Coche, Emmanuel; Vanoverschelde, Jean-Louis J; Gerber, Bernhard L

    2007-01-01

    The purpose of this study was to assess the diagnostic accuracy of 16-slice multidetector-row computed tomography (MDCT) for detecting in-stent restenosis. Fifty patients with 69 previously implanted coronary stents underwent 16-slice MDCT before quantitative coronary angiography (QCA). Diagnostic accuracy of MDCT for detection of in-stent restenosis defined as >50% lumen diameter stenosis (DS) in stented and nonstented coronary segments >1.5-mm diameter was computed using QCA as reference. According to QCA, 18/69 (25%) stented segments had restenosis. In addition, 33/518 (6.4%) nonstented segments had >50% DS. In-stent restenosis was correctly identified on MDCT images in 12/18 stents, and absence of restenosis was correctly identified in 50/51 stents. Stenosis in native coronary arteries was correctly identified in 22/33 segments and correctly excluded in 482/485 segments. Thus, sensitivity (67% vs 67% p=1.0), specificity (98% vs 99%, p=0.96) and overall diagnostic accuracy (90% vs 97%, p=0.68) was similarly high for detecting in-stent restenosis as for detecting stenosis in nonstented coronary segments. MDCT has similarly high diagnostic accuracy for detecting in-stent restenosis as for detecting coronary artery disease in nonstented segments. This suggests that MDCT could be clinically useful for identification of restenosis in patients after coronary stenting.

  10. Diagnostic accuracy of 16-slice multidetector-row CT for detection of in-stent restenosis vs detection of stenosis in nonstented coronary arteries

    Energy Technology Data Exchange (ETDEWEB)

    Kefer, Joelle M.; Vanoverschelde, Jean-Louis J.; Gerber, Bernhard L. [Cliniques Universitaires St. Luc UCL, Department of Cardiology, Brussels (Belgium); Coche, Emmanuel [Cliniques Universitaires St. Luc UCL, Department of Radiology, Brussels (Belgium)

    2007-01-15

    The purpose of this study was to assess the diagnostic accuracy of 16-slice multidetector-row computed tomography (MDCT) for detecting in-stent restenosis. Fifty patients with 69 previously implanted coronary stents underwent 16-slice MDCT before quantitative coronary angiography (QCA). Diagnostic accuracy of MDCT for detection of in-stent restenosis defined as >50% lumen diameter stenosis (DS) in stented and nonstented coronary segments >1.5-mm diameter was computed using QCA as reference. According to QCA, 18/69 (25%) stented segments had restenosis. In addition, 33/518 (6.4%) nonstented segments had >50% DS. In-stent restenosis was correctly identified on MDCT images in 12/18 stents, and absence of restenosis was correctly identified in 50/51 stents. Stenosis in native coronary arteries was correctly identified in 22/33 segments and correctly excluded in 482/485 segments. Thus, sensitivity (67% vs 67% p=1.0), specificity (98% vs 99%, p=0.96) and overall diagnostic accuracy (90% vs 97%, p=0.68) was similarly high for detecting in-stent restenosis as for detecting stenosis in nonstented coronary segments. MDCT has similarly high diagnostic accuracy for detecting in-stent restenosis as for detecting coronary artery disease in nonstented segments. This suggests that MDCT could be clinically useful for identification of restenosis in patients after coronary stenting. (orig.)

  11. Diagnostic performance of computed tomography and magnetic resonance imaging for detecting peritoneal metastases: systematic review and meta-analysis.

    Science.gov (United States)

    Laghi, Andrea; Bellini, Davide; Rengo, Marco; Accarpio, Fabio; Caruso, Damiano; Biacchi, Daniele; Di Giorgio, Angelo; Sammartino, Paolo

    2017-01-01

    Primary end point was to assess diagnostic accuracy of computed tomography (CT) and magnetic resonance imaging (MRI) in detecting peritoneal metastases (PM). Secondary end points were determining the diagnostic sensitivity and specificity of CT in detecting PM according to the peritoneal cancer index (PCI), investigating correlations between radiological and surgical PCI, and comparing diagnostic yield of CT versus positron emission tomography (PET)/CT. We searched MEDLINE, Cochrane Library, Embase and Web of Science databases. Analytic methods were based on PRISMA. Pooled estimates for sensitivity, specificity, positive and negative likelihood ratios were calculated using fixed and random effect models. I (2) was used to evaluate heterogeneity. Of the 529 articles initially identified, 22 were selected for inclusion (934 patients). Cumulative data for per patient CT diagnostic accuracy were sensitivity 83 % (95 % CI 79-86 %), specificity 86 % (95 % CI 82-89 %), pooled positive LR 4.37 (2.58-7.41), and pooled negative LR 0.20 (0.11-0.35). On a per region basis CT performed best in epigastrium and pelvis. Correlation analysis showed a high correlation between CT-PCI and surgical-PCI scores, ranging from 0.49 to 0.96. MRI and PET/CT achieved similar per patient diagnostic accuracy. CT should be the preferred diagnostic imaging modality for detecting peritoneal metastases because of the robustness of the data. MRI and PET/CT should be considered second choices, until more consistent information on their diagnostic yield in detecting PM are obtained.

  12. New technology for ultrasensitive detection and isolation of rare cells for clinical diagnostics and therapeutics

    Science.gov (United States)

    Leary, James F.; McLaughlin, Scott R.

    1995-04-01

    A high-speed, 11-parameter, 6-color fluorescence, laser flow cytometer/cell sorter with a number of special and unique features has been built for ultrasensitive detection and isolation of rare cells for clinical diagnostics and therapeutics. The software for real-time data acquisition and sort control, written as C++ programming language modules with a WindowsTM graphical user interface, runs on a 66-MHz 80486 computer joined by an extended bus to 23 sophisticated multi-layered boards of special data acquisition and sorting electronics. Special features include: high-speed (> 100,000 cells/sec) real-time data classification module (U.S. Patent 5,204,884 (1993)); real-time principal component cell sorting; multi-queue signal-processing system with multiple hardware and software event buffers to reduce instrument dead time, LUT charge-pulse definition, high-resolution `flexible' sorting for optimal yield/purity sort strategies (U.S. Patent 5,199,576); pre-focusing optical wavelength correction for a second laser beam; and two trains of three fluorescence detectors-- each adjustable for spatial separation to interrogate only one of two laser beams, syringe- driven or pressure-driven fluidics, and time-windowed parameters. The system has been built to be both expandable and versatile through the use of LUT's and a modular hardware and software design. The instrument is especially useful at detection and isolation of rare cell subpopulations for which our laboratory is well-known. Cell subpopulations at frequencies as small as 10-7 have been successfully studied with this system. Current applications in clinical diagnostics and therapeutics include detection and isolation of (1) fetal cells from material blood for prenatal diagnosis of birth defects, (2) hematopoietic stem and precursor cells for autologous bone marrow transplantation, (3) metastatic breast cancer cells for molecular characterization, and (4) HIV-infected maternal cells in newborn blood to study mother

  13. Comparative efficacy of conventional primer sets in detection of Cryptosporidium parvum for diagnostic use.

    Science.gov (United States)

    Kar, Sirri; Daugschies, Arwid; Bangoura, Berit

    2010-02-01

    In this study, the sensitivity and specificity of different previously described primer sets for Cryptosporidium parvum detection by polymerase chain reaction (PCR) was evaluated. For this purpose, the primer sets defined by Cacciò et al. (FEMS Microbiol Lett 170(1):173-179, 1999) (tub), Widmer et al. (Appl Environ Microbiol 64(11):4477-4481, 1998) (btub) and Rochelle et al. (Appl Environ Microbiol 63:2029-2037, 1997) (cphsp), respectively, were used. Deoxyribonucleic acid (DNA) was isolated from three different sample materials: (1) from the faeces of an experimentally C. parvum-infected calf, (2) from purified C. parvum oocysts, and (3) from C. parvum-infected HCT-8 cell cultures. The DNA samples were subjected to PCR reactions with each of the three given primer sets to investigate sensitivity and suitability for routine use. The primers described by Cacciò et al. (FEMS Microbiol Lett 170(1):173-179, 1999) (TUB) were superior regarding sensitivity and specificity in terms of detection of C. parvum in faeces, in purified oocysts and also in cell culture, and may thus be applied for routine diagnostic use in common sample materials.

  14. Automated detection of diagnostically relevant regions in H&E stained digital pathology slides

    Science.gov (United States)

    Bahlmann, Claus; Patel, Amar; Johnson, Jeffrey; Ni, Jie; Chekkoury, Andrei; Khurd, Parmeshwar; Kamen, Ali; Grady, Leo; Krupinski, Elizabeth; Graham, Anna; Weinstein, Ronald

    2012-03-01

    We present a computationally efficient method for analyzing H&E stained digital pathology slides with the objective of discriminating diagnostically relevant vs. irrelevant regions. Such technology is useful for several applications: (1) It can speed up computer aided diagnosis (CAD) for histopathology based cancer detection and grading by an order of magnitude through a triage-like preprocessing and pruning. (2) It can improve the response time for an interactive digital pathology workstation (which is usually dealing with several GByte digital pathology slides), e.g., through controlling adaptive compression or prioritization algorithms. (3) It can support the detection and grading workflow for expert pathologists in a semi-automated diagnosis, hereby increasing throughput and accuracy. At the core of the presented method is the statistical characterization of tissue components that are indicative for the pathologist's decision about malignancy vs. benignity, such as, nuclei, tubules, cytoplasm, etc. In order to allow for effective yet computationally efficient processing, we propose visual descriptors that capture the distribution of color intensities observed for nuclei and cytoplasm. Discrimination between statistics of relevant vs. irrelevant regions is learned from annotated data, and inference is performed via linear classification. We validate the proposed method both qualitatively and quantitatively. Experiments show a cross validation error rate of 1.4%. We further show that the proposed method can prune ~90% of the area of pathological slides while maintaining 100% of all relevant information, which allows for a speedup of a factor of 10 for CAD systems.

  15. A novel diagnostic aid for intra-abdominal adhesion detection in cine-MR imaging: Pilot study and initial diagnostic impressions.

    Science.gov (United States)

    Randall, David; Joosten, Frank; ten Broek, Richard; Gillott, Richard; Bardhan, Karna Dev; Strik, Chema; Prins, Wiesje; van Goor, Harry; Fenner, John

    2017-07-14

    A non-invasive diagnostic technique for abdominal adhesions is not currently available. Capture of abdominal motion due to respiration in cine-MRI has shown promise, but is difficult to interpret. This article explores the value of a complimentary diagnostic aid to facilitate the non-invasive detection of abdominal adhesions using cine-MRI. An image processing technique was developed to quantify the amount of sliding that occurs between the organs of the abdomen and the abdominal wall in sagittal cine-MRI slices. The technique produces a 'sheargram' which depicts the amount of sliding which has occurred over 1-3 respiratory cycles. A retrospective cohort of 52 patients, scanned for suspected adhesions, made 281 cine-MRI sagittal slices available for processing. The resulting sheargrams were reported by two operators and compared to expert clinical judgement of the cine-MRI scans. The sheargram matched clinical judgement in 84% of all sagittal slices and 93-96% of positive adhesions were identified on the sheargram. The sheargram displayed a slight skew towards sensitivity over specificity, with a high positive adhesion detection rate but at the expense of false positives. Good correlation between sheargram and absence/presence of inferred adhesions indicates quantification of sliding motion has potential to aid adhesion detection in cine-MRI. Advances in Knowledge: This is the first attempt to clinically evaluate a novel image processing technique quantifying the sliding motion of the abdominal contents against the abdominal wall. The results of this pilot study reveal its potential as a diagnostic aid for detection of abdominal adhesions.

  16. Kinetics of hybridization on surface oligonucleotide microchips: theory, experiment, and comparison with hybridization on gel-based microchips.

    Science.gov (United States)

    Sorokin, N V; Chechetkin, V R; Pan'kov, S V; Somova, O G; Livshits, M A; Donnikov, M Y; Turygin, A Y; Barsky, V E; Zasedatelev, A S

    2006-08-01

    The optimal design of oligonucleotide microchips and efficient discrimination between perfect and mismatch duplexes strongly depend on the external transport of target DNA to the cells with immobilized probes as well as on respective association and dissociation rates at the duplex formation. In this paper we present the relevant theory for hybridization of DNA fragments with oligonucleotide probes immobilized in the cells on flat substrate. With minor modifications, our theory also is applicable to reaction-diffusion hybridization kinetics for the probes immobilized on the surface of microbeads immersed in hybridization solution. The main theoretical predictions are verified with control experiments. Besides that, we compared the characteristics of the surface and gel-based oligonucleotide microchips. The comparison was performed for the chips printed with the same pin robot, for the signals measured with the same devices and processed by the same technique, and for the same hybridization conditions. The sets of probe oligonucleotides and the concentrations of probes in respective solutions used for immobilization on each platform were identical as well. We found that, despite the slower hybridization kinetics, the fluorescence signals and mutation discrimination efficiency appeared to be higher for the gel-based microchips with respect to their surface counterparts even for the relatively short hybridization time about 0.5-1 hour. Both the divergence between signals for perfects and the difference in mutation discrimination efficiency for the counterpart platforms rapidly grow with incubation time. In particular, for hybridization during 3 h the signals for gel-based microchips surpassed their surface counterparts in 5-20 times, while the ratios of signals for perfect-mismatch pairs for gel microchips exceeded the corresponding ratios for surface microchips in 2-4 times. These effects may be attributed to the better immobilization efficiency and to the higher

  17. A novel fully automated molecular diagnostic system (AMDS for colorectal cancer mutation detection.

    Directory of Open Access Journals (Sweden)

    Shiro Kitano

    Full Text Available BACKGROUND: KRAS, BRAF and PIK3CA mutations are frequently observed in colorectal cancer (CRC. In particular, KRAS mutations are strong predictors for clinical outcomes of EGFR-targeted treatments such as cetuximab and panitumumab in metastatic colorectal cancer (mCRC. For mutation analysis, the current methods are time-consuming, and not readily available to all oncologists and pathologists. We have developed a novel, simple, sensitive and fully automated molecular diagnostic system (AMDS for point of care testing (POCT. Here we report the results of a comparison study between AMDS and direct sequencing (DS in the detection of KRAS, BRAF and PI3KCA somatic mutations. METHODOLOGY/PRINCIPAL FINDING: DNA was extracted from a slice of either frozen (n = 89 or formalin-fixed and paraffin-embedded (FFPE CRC tissue (n = 70, and then used for mutation analysis by AMDS and DS. All mutations (n = 41 among frozen and 27 among FFPE samples detected by DS were also successfully (100% detected by the AMDS. However, 8 frozen and 6 FFPE samples detected as wild-type in the DS analysis were shown as mutants in the AMDS analysis. By cloning-sequencing assays, these discordant samples were confirmed as true mutants. One sample had simultaneous "hot spot" mutations of KRAS and PIK3CA, and cloning assay comfirmed that E542K and E545K were not on the same allele. Genotyping call rates for DS were 100.0% (89/89 and 74.3% (52/70 in frozen and FFPE samples, respectively, for the first attempt; whereas that of AMDS was 100.0% for both sample sets. For automated DNA extraction and mutation detection by AMDS, frozen tissues (n = 41 were successfully detected all mutations within 70 minutes. CONCLUSIONS/SIGNIFICANCE: AMDS has superior sensitivity and accuracy over DS, and is much easier to execute than conventional labor intensive manual mutation analysis. AMDS has great potential for POCT equipment for mutation analysis.

  18. A preface on advances in diagnostics for infectious and parasitic diseases: detecting parasites of medical and veterinary importance.

    Science.gov (United States)

    Stothard, J Russell; Adams, Emily

    2014-12-01

    There are many reasons why detection of parasites of medical and veterinary importance is vital and where novel diagnostic and surveillance tools are required. From a medical perspective alone, these originate from a desire for better clinical management and rational use of medications. Diagnosis can be at the individual-level, at close to patient settings in testing a clinical suspicion or at the community-level, perhaps in front of a computer screen, in classification of endemic areas and devising appropriate control interventions. Thus diagnostics for parasitic diseases has a broad remit as parasites are not only tied with their definitive hosts but also in some cases with their vectors/intermediate hosts. Application of current diagnostic tools and decision algorithms in sustaining control programmes, or in elimination settings, can be problematic and even ill-fitting. For example in resource-limited settings, are current diagnostic tools sufficiently robust for operational use at scale or are they confounded by on-the-ground realities; are the diagnostic algorithms underlying public health interventions always understood and well-received within communities which are targeted for control? Within this Special Issue (SI) covering a variety of diseases and diagnostic settings some answers are forthcoming. An important theme, however, throughout the SI is to acknowledge that cross-talk and continuous feedback between development and application of diagnostic tests is crucial if they are to be used effectively and appropriately.

  19. Imaging strategies for detection of urgent conditions in patients with acute abdominal pain: diagnostic accuracy study

    Science.gov (United States)

    Laméris, Wytze; van Randen, Adrienne; van Es, H Wouter; van Heesewijk, Johannes P M; van Ramshorst, Bert; Bouma, Wim H; ten Hove, Wim; van Leeuwen, Maarten S; van Keulen, Esteban M; Dijkgraaf, Marcel G W; Bossuyt, Patrick M M; Boermeester, Marja A

    2009-01-01

    Objective To identify an optimal imaging strategy for the accurate detection of urgent conditions in patients with acute abdominal pain. Design Fully paired multicentre diagnostic accuracy study with prospective data collection. Setting Emergency departments of two university hospitals and four large teaching hospitals in the Netherlands. Participants 1021 patients with non-traumatic abdominal pain of >2 hours’ and <5 days’ duration. Exclusion criteria were discharge from the emergency department with no imaging considered warranted by the treating physician, pregnancy, and haemorrhagic shock. Intervention All patients had plain radiographs (upright chest and supine abdominal), ultrasonography, and computed tomography (CT) after clinical and laboratory examination. A panel of experienced physicians assigned a final diagnosis after six months and classified the condition as urgent or non-urgent. Main outcome measures Sensitivity and specificity for urgent conditions, percentage of missed cases and false positives, and exposure to radiation for single imaging strategies, conditional imaging strategies (CT after initial ultrasonography), and strategies driven by body mass index and age or by location of pain. Results 661 (65%) patients had a final diagnosis classified as urgent. The initial clinical diagnosis resulted in many false positive urgent diagnoses, which were significantly reduced after ultrasonography or CT. CT detected more urgent diagnoses than did ultrasonography: sensitivity was 89% (95% confidence interval 87% to 92%) for CT and 70% (67% to 74%) for ultrasonography (P<0.001). A conditional strategy with CT only after negative or inconclusive ultrasonography yielded the highest sensitivity, missing only 6% of urgent cases. With this strategy, only 49% (46% to 52%) of patients would have CT. Alternative strategies guided by body mass index, age, or location of the pain would all result in a loss of sensitivity. Conclusion Although CT is the most

  20. Class imbalance in unsupervised change detection - A diagnostic analysis from urban remote sensing

    Science.gov (United States)

    Leichtle, Tobias; Geiß, Christian; Lakes, Tobia; Taubenböck, Hannes

    2017-08-01

    Automatic monitoring of changes on the Earth's surface is an intrinsic capability and simultaneously a persistent methodological challenge in remote sensing, especially regarding imagery with very-high spatial resolution (VHR) and complex urban environments. In order to enable a high level of automatization, the change detection problem is solved in an unsupervised way to alleviate efforts associated with collection of properly encoded prior knowledge. In this context, this paper systematically investigates the nature and effects of class distribution and class imbalance in an unsupervised binary change detection application based on VHR imagery over urban areas. For this purpose, a diagnostic framework for sensitivity analysis of a large range of possible degrees of class imbalance is presented, which is of particular importance with respect to unsupervised approaches where the content of images and thus the occurrence and the distribution of classes are generally unknown a priori. Furthermore, this framework can serve as a general technique to evaluate model transferability in any two-class classification problem. The applied change detection approach is based on object-based difference features calculated from VHR imagery and subsequent unsupervised two-class clustering using k-means, genetic k-means and self-organizing map (SOM) clustering. The results from two test sites with different structural characteristics of the built environment demonstrated that classification performance is generally worse in imbalanced class distribution settings while best results were reached in balanced or close to balanced situations. Regarding suitable accuracy measures for evaluating model performance in imbalanced settings, this study revealed that the Kappa statistics show significant response to class distribution while the true skill statistic was widely insensitive to imbalanced classes. In general, the genetic k-means clustering algorithm achieved the most robust results

  1. An In Vitro Comparison of Different Diagnostic Methods in Detection of Residual Dentinal Caries

    Directory of Open Access Journals (Sweden)

    Nimet Unlu

    2010-01-01

    Full Text Available The aim of this study was to investigate the efficiency of different diagnostic methods in detection of residual dentinal caries in excavated cavities. Fifty extracted molar with deep dentinal carious lesions were excavated using a slow-speed handpiece. All cavities were assessed by laser fluorescence(LF device, electronic caries monitor(ECM, and caries detector dye(CDD by three independent observers blindly. The measurements were repeated after two weeks. Specimens containing dentin slices 150 μm in thickness were prepared for histological analyses. The existence and absence of carious dentin was determined using a lightmicroscope. The average intraobserver accuracy was 1.00 (perfect agreement for CDD, 0.86 (excellent agreement for ECM, and 0.50 (good agreement for LF. The average interobserver accuracy values were 0.92 (excellent agreement, (0.36 marginal agreement and 0.48 (good agreement, for CDD, ECM, and LF, respectively. The average specificity was 0.60 for CDD, 73% for ECM, and 0.50 for LF. The average sensitivity was 0.55 for CDD, 0.85 for LF, and 0.47 for ECM. The average accuracy values were 0.53, 0.51, and 0.81 for CDD, ECM, and LF, respectively. LF had the greatest sensitivity and accuracy values of any of the methods tested. As a conclusion, LF device is appeared to most reliable method in detection of remain caries in cavity. However, because of its technical sensitivity it may susceptible to variations in measurements. To pay attention to the rule of usage and repeated measurements can minimize such variations in clinical practice. It was concluded that LF is an improvement on the currently available aids for residual caries detection.

  2. A Multiplexed Diagnostic Platform for Point-of-Care Pathogen Detection

    Energy Technology Data Exchange (ETDEWEB)

    Regan, J F; Letant, S E; Adams, K L; Mahnke, R C; Nguyen, N T; Dzenitis, J M; Hindson, B J; Hadley, D R; Makarewicz, T J; Henderer, B D; Breneman, J W; Tammero, L F; Ortiz, J I; Derlet, R W; Cohen, S; Colston, W W; McBride, M T; Birch, J M

    2008-02-04

    We developed an automated point-of-care diagnostic instrument that is capable of analyzing nasal swab samples for the presence of respiratory diseases. This robust instrument, called FluIDx, performs autonomous multiplexed RT-PCR reactions that are analyzed by microsphere xMAP technology. We evaluated the performance of FluIDx, in comparison rapid tests specific for influenza and respiratory syncytial virus, in a clinical study performed at the UC Davis Medical Center. The clinical study included samples positive for RSV (n = 71), influenza A (n = 16), influenza B (n = 4), adenovirus (n = 5), parainfluenza virus (n = 2), and 44 negative samples, according to a composite reference method. FluIDx and the rapid tests detected 85.9% and 62.0% of the RSV positive samples, respectively. Similar sensitivities were recorded for the influenza B samples; whereas the influenza A samples were poorly detected, likely due to the utilization of an influenza A signature that did not accurately match currently circulating influenza A strains. Data for all pathogens were compiled and indicate that FluIDx is more sensitive than the rapid tests, detecting 74.2% (95% C.I. of 64.7-81.9%) of the positive samples in comparison to 53.6% (95% C.I. of 43.7-63.2%) for the rapid tests. The higher sensitivity of FluIDx was partially offset by a lower specificity, 77.3% versus 100.0%. Overall, these data suggest automated flow-through PCR-based instruments that perform multiplexed assays can successfully screen clinical samples for infectious diseases.

  3. Detection of Hyperechoic Inflammatory Fatty Tissue during Transabdominal Ultrasonography: Diagnostic Role in Acute Abdomen

    Energy Technology Data Exchange (ETDEWEB)

    Park, Seong Jin; Lee, Hae Kyung; Yi, Bum Ha [Soonchunhyang University Bucheon Hospital, Bucheon (Korea, Republic of); Kim, Hyun Cheol [Soonchunhyang University Cheonan Hospital, Cheonan (Korea, Republic of)

    2005-12-15

    To assess the incidence and diagnostic role of hyperechoic inflammatory fatty tissue (HIFT) in transabdominal ultrasonography (TAUS) for acute abdomen. With TAUS, we examined 98 consecutive patients (68 women, 30 men: mean age, 32 years: age range, 4-84 years) having acute abdominal pain. We examined the abdomen and pelvis by TAUS to determine the cause of acute abdomen, to check for the presence of HIFT, and to investigate whether it was easier and earlier to find the main cause and HIFT presence. We also prospectively evaluated the shape, distribution, and diagnostic role of HIFT. Final diagnoses consisted of 47 cases of acute appendicitis, 14 of enterocolitis, 13 of PID, 7 of gynecological hemoperitoneum, 5 of colonic diverticulitis, 3 of ovarian torsion, 2 of colon perforation, 2 of only presence of non-specific HIFT, 1 of mesenteric lymphadenitis, and 4 of normal. HIFT were seen in 67 patients (68.4%), including 44/47(93.6%) of acute appendicitis, 2/14(14.3%) of enterocolitis, 11/13(84.6%) of PID, 0/7 of hemoperitoneum, 5/5 of colonic diverticulitis, 0/3 of ovarian torsion, 2/2 of colon perforation, and 1/1 mesenteric lymphadenitis. HIFT were detected earlier than the main cause in 17/44 of acute appendicitis, 6/11 of PID, and 4/5 of colonic diverticulitis. In acute appendicitis, the shape of HIFT appeared as fat thickening along the mesoappendix in 12/44, fat thickening along the mesoappendix and the opposite side in 13/44, fat encircled appendix in 6/44, fatty mass wrapping abscess in 10/44, and diffuse intraperitoneal fat thickening in 3/44. In PID, HIFT appeared as a single fatty mass in the pelvis and lower abdomen in 6/11, wrapping pelvic abscess in 2/11, and multiple fatty masses scattered in abdomen and pelvis in 3/11. In colonic diverticulitis, all 5 cases appeared as hyperechoic hemispheric mass covering the inflamed diverticulum. HIFT are a usual US finding in patients with acute abdomen, particularly on abdominal and pelvic inflammatory conditions

  4. Diagnostic accuracy of referral criteria for head circumference to detect hydrocephalus in the first year of life

    NARCIS (Netherlands)

    Dommelen, P. van; Deurloo, J.A.; Gooskens, R.H.; Verkerk, P.H.

    2015-01-01

    Background Increased head circumference is often the first and main sign leading to the diagnosis of hydrocephalus. Our aim is to investigate the diagnostic accuracy of referral criteria for head circumference to detect hydrocephalus in the first year of life. Methods A reference group with longitud

  5. Diagnostic accuracy of referral criteria for head circumference to detect hydrocephalus in the first year of life

    NARCIS (Netherlands)

    Dommelen, P. van; Deurloo, J.A.; Gooskens, R.H.; Verkerk, P.H.

    2015-01-01

    Background Increased head circumference is often the first and main sign leading to the diagnosis of hydrocephalus. Our aim is to investigate the diagnostic accuracy of referral criteria for head circumference to detect hydrocephalus in the first year of life. Methods A reference group with

  6. A comparative study of the diagnostic accuracy on Waters view with CT scan in detecting midface fractures

    OpenAIRE

    Panjnoush M.; Shirani Gh.; Jozghanbari P.

    2006-01-01

    Background and Aim: In recent years, CT scan has become available as an alternative to conventional radiography. To date, the utility of Waters view in detecting midface fractures has been rarely evaluated. The aim of this study was to compare the diagnostic accuracy and reliability of Waters radiography with CT scan in detecting midface fractures. Materials and Methods: In this tests evaluation study, waters view and CT scan were performed for 42 patients with midface fracture admitted to ma...

  7. Longevity of radiofrequency identification device microchips in citrus trees

    Science.gov (United States)

    Long-term identification of individual plants in the field is an important part of many types of botanical and horticultural research. In a previous report, we described methods for using implanted radiofrequency (RFID) microchips to tag citrus trees for field research. This report provides an upd...

  8. Apparatus and method for performing electrodynamic focusing on a microchip

    Science.gov (United States)

    Ramsey, John Michael; Jacobson, Stephen C.

    1999-01-01

    A microchip device includes a focusing channel, in which an electric field strength established in the focusing channel is controlled relative to an electric field strength established in a material transport channel segment to spatially focus the material traversing the material transport channel segment.

  9. Transverse mode selection in a monolithic microchip laser

    CSIR Research Space (South Africa)

    Naidoo, Darryl

    2011-11-01

    Full Text Available The article outlines an approach to mode selection in a microchip laser through judicious shaping of the pump light to create a high modal overlap with the desired mode. The authors demonstrate the principle by creating a donut-shaped pump profile...

  10. Dynamics of chaotic oscillations in mutually coupled microchip lasers

    CERN Document Server

    Uchida, A; Kinugawa, S; Yoshimori, S

    2003-01-01

    We have numerically and experimentally investigated the dynamics of mutually coupled microchip lasers. Chaotic oscillations are observed in the vicinity of the boundary of the injection-locking range when the coupling strength and the difference of the optical frequencies are varied. Synchronization of chaos is always achieved under the condition to generate chaos.

  11. Carbon Nanotube-Based Separation Columns for Microchip Electrochromatography

    DEFF Research Database (Denmark)

    Mogensen, Klaus Bo; Delacourt, B.; Kutter, Jörg Peter

    2015-01-01

    Fabrication of the stationary phase for microchip chromatography is most often done by packing of the individual separation channel after fabrication of the microfluidic chip, which is a very time-consuming and costly process (Kutter. J Chromatogr A 1221:72–82, 2012). Here, we describe in detail...

  12. Problems Associated with the Microchip Data of Stray Dogs and Cats Entering RSPCA Queensland Shelters

    Directory of Open Access Journals (Sweden)

    Emily Lancaster

    2015-05-01

    Full Text Available A lack of published information documenting problems with the microchip data for the reclaiming of stray animals entering Australian shelters limits improvement of the current microchipping system. A retrospective study analysing admission data for stray, adult dogs (n = 7258 and cats (n = 6950 entering the Royal Society for the Prevention of Cruelty to Animals (RSPCA Queensland between January 2012 and December 2013 was undertaken to determine the character and frequency of microchip data problems and their impact on outcome for the animal. Only 28% of dogs and 9% of cats were microchipped, and a substantial proportion (37% had problems with their data, including being registered to a previous owner or organisation (47%, all phone numbers incorrect/disconnected (29%, and the microchip not registered (14%. A higher proportion of owners could be contacted when the microchip had no problems, compared to those with problems (dogs, 93% vs. 70%; cats, 75% vs. 41%. The proportion of animals reclaimed declined significantly between microchipped animals with no data problems, microchipped animals with data problems and non-microchipped animals—87%, 69%, and 37%, respectively, for dogs and 61%, 33%, and 5%, respectively, for cats. Strategies are needed to increase the accuracy of microchip data to facilitate the reclaiming of stray dogs and cats.

  13. Comparing polyelectrolyte multilayer-coated PMMA microfluidic devices and glass microchips for electrophoretic separations.

    Science.gov (United States)

    Currie, Christa A; Shim, Joon Sub; Lee, Se Hwan; Ahn, Chong; Limbach, Patrick A; Halsall, H Brian; Heineman, William R

    2009-12-01

    There is a continuing drive in microfluidics to transfer microchip systems from the more expensive glass microchips to cheaper polymer microchips. Here, we investigate using polyelectrolyte multilayers (PEM) as a coating system for PMMA microchips to improve their functionality. The multilayer system was prepared by layer-to-layer deposition of poly(diallyldimethylammonium) chloride and polystyrene sulfonate. Practical aspects of coating PMMA microchips were explored. The multilayer buildup process was monitored using EOF measurements, and the stability of the PEM was investigated. The performance of the PEM-PMMA microchip was compared with those of a standard glass microchip and a PEM-glass microchip in terms of EOF and separating two fluorescent dyes. Several key findings in the development of the multilayer coating procedure for PMMA chips are also presented. It was found that, with careful preparation, a PEM-PMMA microchip can be prepared that has properties comparable--and in some cases superior--to those of a standard glass microchip.

  14. The diagnostic value of sacroiliac CT for detecting early changes of ankylosing spondylitis

    Energy Technology Data Exchange (ETDEWEB)

    Park, Ju Hyun; Park, Ji Seon; Ryu, Kyung Nam [Kyunghee University Medical Center, Seoul (Korea, Republic of); Jin, Wook [East-West Neo Medical Center, Seoul (Korea, Republic of)

    2007-02-15

    We wanted to evaluate the diagnostic value of the various findings on sacroiliac CT for detecting the early changes of ankylosing spondylitis (AS). Between April 2005 and March 2006, 51 sacroiliac CT images with the clinical suspicion of AS, but no definite evidence of AS on the plain radiograph only, were retrospectively reviewed. Finally, 36 patients (mean age: 28.6 years; 34 males and 2 females), who were clinically diagnosed as AS (AS group, n = 26) or they had no evidence of AS (non-AS group, n = 10), were evaluated. Two musculoskeletal radiologists analyzed the presence of marginal irregularity, bony erosion and subchondral sclerosis. A statistical analysis was performed to evaluate the incidence, sensitivity and specificity of each finding. Marginal irregularity was seen in 21 of 26 AS subjects, and in 8 of 10 non-AS subjects. Bony erosion was only seen in 13 of 26 AS subjects. Subchondral sclerosis was observed in 19 of 26 AS subjects and in 8 of 10 non-AS subjects. The sensitivity/specificity for each findings were 72.4%/28.6%, 100%/43.5% and 70.4%/22.2%, respectively. Except for bony erosions, these results showed no statistical significance ({rho} = .006). Bony erosion on CT is a very sensitive finding for the early changes of AS, whereas marginal irregularity or subchondral sclerosis is not so helpful in differentiating AS from non-AS. Attention to these results may further enhance the accurate diagnosis of the early changes in AS.

  15. Field evaluation of a rapid diagnostic test to detect antibodies in human toxocariasis.

    Science.gov (United States)

    Lim, P K C; Yamasaki, H; Mak, J W; Wong, S F; Chong, C W; Yap, I K S; Ambu, S; Kumarasamy, V

    2015-08-01

    Human toxocariasis which is caused mainly by the larvae of Toxocara canis and Toxocara cati, is a worldwide zoonotic disease that can be a potentially serious human infection. The enzyme-linked immunosorbent assay (ELISA) using T. canis excretory-secretory (TES) antigens harvested from T. canis larvae is currently the serological test for confirming toxocariasis. An alternative to producing large amounts of Toxocara TES and improved diagnosis for toxocariasis is through the development of highly specific recombinant antigens such as the T. canis second stage larva excretory-secretory 30 kDa protein (recTES-30). The aim of this study was to evaluate the sensitivity and specificity of a rapid diagnostic kit (RDT, named as iToxocara kit) in comparison to recTES-30 ELISA in Serendah Orang Asli village in Selangor, Malaysia. A total of 133 subjects were included in the study. The overall prevalence rates by ELISA and RDT were 29.3% and 33.1%, respectively, with more positive cases detected in males than females. However, no association was found between toxocariasis and gender or age. The percentage sensitivity, specificity, positive predictive value and negative predictive value of RDT were 85.7%, 90.1%, 80% and 93.2%, respectively. The prevalence for toxocariasis in this population using both ELISA and RDT was 27.1% (36/133) and the K-concordance test suggested good agreement of the two tests with a Cohen's kappa of 0.722, Ptoxocariasis.

  16. Wavelet diagnostics for detection of coherent structures in instantaneous turbulent flow imagery: A review

    Indian Academy of Sciences (India)

    Roddam Narasimha

    2007-02-01

    A review of work over the last decade shows that 2D wavelet techniques applied on flow imagery can provide powerful insights into the nature and lifecycle of coherent structures (the latter through wavelet movies) in turbulent shear flows. The advantage of wavelet techniques in often being able to infer the nature of coherent motion from a single image is emphasized. The techniques are first calibrated by using them on well-known results in the turbulent mixing layer. They are then applied to jets and plumes, and it is shown how off-source heating in such flows can disrupt the coherent structures in the unheated flow. A suitably reduced version of the present method, using discrete wavelet transforms on signals from a finite array of sensors, could be a useful diagnostic tool in near-real-time detection of coherent structures or patterns for the purpose of selecting appropriate control signals to the actuators in a flow-control system.

  17. Point-of-care Diagnostic Tools to Detect Circulating MicroRNAS as Biomarkers of Disease

    Directory of Open Access Journals (Sweden)

    Luis Vaca

    2014-05-01

    Full Text Available MicroRNAs or miRNAs are a form of small non-coding RNAs (ncRNAs of 19–22 nucleotides in length in their mature form. miRNAs are transcribed in the nucleus of all cells from large precursors, many of which have several kilobases in length. Originally identified as intracellular modulators of protein synthesis via posttranscriptional gene silencing, more recently it has been found that miRNAs can travel in extracellular human fluids inside specialized vesicles known as exosomes. We will be referring to this miRNAs as circulating microRNAs. More interestingly, the miRNA content inside exosomes changes during pathological events. In the present review we analyze the literature about circulating miRNAs and their possible use as biomarkers. Furthermore, we explore their future in point-of-care (POC diagnostics and provide an example of a portable POC apparatus useful in the detection of circulating miRNAs.

  18. Raman spectroscopy: a diagnostic tool for detection of early malignant changes in the larynx

    Science.gov (United States)

    Stone, Nicholas; Stavroulaki, Pelagia; Kendall, Catherine A.; Birchall, Martin; Barr, Hugh

    2000-05-01

    The incidence of laryngeal cancer has risen progressively over the last 25 years. Early diagnosis and treatment of premalignant lesions of the larynx is vital to prevent progression to invasive squamous cell carcinoma. In the larynx, it has long been recognized that histological evidence of maturation abnormality is associated with a higher risk of transformation to malignancy. Currently, it is extremely difficult if not impossible for the clinician to ascertain the level of abnormality present without removing a biopsy sample and sending it for histopathological analysis. Inherent risks with this technique include damage to vocal chords and loss of speech quality as well as possible selection of unrepresentative biopsy samples. Raman spectroscopy, incorporated into an endoscopic system, has the potential to provide a real-time, non-invasive diagnostic technique able to detect biochemical changes that accompany abnormal pathology. Likely outcomes would be improved biopsy targeting and patient management by providing immediate result of tissue pathology. This paper demonstrates the capacity of near IR Raman spectroscopy combine with statistical data analysis techniques to discriminate between normal, dysplastic and cancerous laryngeal tissue.

  19. MDA-9 and GRP78 as potential diagnostic biomarkers for early detection of melanoma metastasis.

    Science.gov (United States)

    Guan, Ming; Chen, Xiaofan; Ma, Yingyu; Tang, Lihua; Guan, Lei; Ren, Xuefeng; Yu, Bo; Zhang, Wei; Su, Bing

    2015-04-01

    Metastatic melanoma, the primary cause of skin cancer-related death, warrants new diagnostic and therapeutic approaches that target the regulatory machinery at molecular level. The heterogeneity and complexity of melanoma result in the difficulty to find biomarkers and targets for early detection and treatment. Here, we investigated metastasis-associated proteins by comparing the proteomic profiles of primary cutaneous melanomas to their matched lymph node metastases, which minimizes heterogeneity among samples from different patients. Results of two-dimensional gel electrophoresis (2-DE) followed by proteomic analysis revealed eight differentially expressed proteins. Among them, seven proteins (α-enolase, cofilin-1, LDH, m-β-actin, Nm23, GRP78, and MDA-9) showed increased and one (annexin A2) showed decreased expression in metastatic lymph node tissues than in primary melanomas. MDA-9 and GRP78 were the most highly expressed proteins in lymph node metastases, which was validated by immunohistochemical staining. Moreover, exosomes from serum samples of metastatic melanoma patients contained higher levels of MDA-9 and GRP78 than those of patients without metastases, indicating the potential of MDA-9 and GRP78 to be biomarkers for early detection of metastasis. Further, small interfering RNA (siRNA)-mediated knockdown confirmed a functional role for MDA-9 and GRP78 to promote cell invasion in the A375 cells. Finally, we showed that GRP78 co-localized with MDA-9 in 293T cells. Taken together, our findings support MDA-9, co-expressed with GRP78, as a melanoma protein associated with lymph node metastasis. Investigating how MDA-9 and GRP78 interact to contribute to melanoma metastasis and disease progression could reveal new potential avenues of targeted therapy and/or useful biomarkers for diagnosis and prognosis.

  20. Rapid Real-Time Antimicrobial Susceptibility Testing with Electrical Sensing on Plastic Microchips with Printed Electrodes.

    Science.gov (United States)

    Safavieh, Mohammadali; Pandya, Hardik J; Venkataraman, Maanasa; Thirumalaraju, Prudhvi; Kanakasabapathy, Manoj Kumar; Singh, Anupriya; Prabhakar, Devbalaji; Chug, Manjyot Kaur; Shafiee, Hadi

    2017-03-30

    Rapid antimicrobial susceptibility testing is important for efficient and timely therapeutic decision making. Due to globally spread bacterial resistance, the efficacy of antibiotics is increasingly being impeded. Conventional antibiotic tests rely on bacterial culture, which is time-consuming and can lead to potentially inappropriate antibiotic prescription and up-front broad range of antibiotic use. There is an urgent need to develop point-of-care platform technologies to rapidly detect pathogens, identify the right antibiotics, and monitor mutations to help adjust therapy. Here, we report a biosensor for rapid (microchips with printed electrodes using antibodies (30 min), and its electrical response is monitored in the presence and absence of antibiotics over an hour of incubation time. We evaluated the microchip with Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA) as clinical models with ampicillin, ciprofloxacin, erythromycin, daptomycin, gentamicin, and methicillin antibiotics. The results are compared with the current standard methods, i.e. bacteria viability and conventional antibiogram assays. The technology presented here has the potential to provide precise and rapid bacteria screening and guidance in clinical therapies by identifying the correct antibiotics for pathogens.

  1. Diagnostic value of the anterior chamber depth of a globe on CT for detecting open-globe injury

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Seong Yun; Lee, Jeong Hyun; Choi, Byung Se; Choi, Jin Woo; In, Hyun Sin; Kim, Sun Mi; Baek, Jung Hwan [University of Ulsan College of Medicine, Asan Medical Centre, Department of Radiology and Research Institute of Radiology, Seoul (Korea); Lee, Yong Jae [University of Ulsan College of Medicine, Asan Medical Centre, Department of Ophthalmology, Seoul (Korea)

    2010-05-15

    To determine the diagnostic value of the anterior chamber depth (ACD) of a globe as seen on CT for detecting open-globe injury. We enrolled 28 patients with unilateral open-globe injury confirmed by surgery and 28 controls with other types of orbital trauma. The diagnostic value of the difference of the ACDs between globes was evaluated by ROC analysis. The diagnostic performance of the following CT findings suggesting open-globe injury was also determined from independent analysis by two readers, i.e. change in the ACD and the globe contour, obvious volume loss of the globe, dislocated/deformed lens, intraocular foreign body/air and intraocular haemorrhage. The ACD measurements were possible only in 15 patients with open-globe injuries. A difference in the ACDs{>=}0.4 mm resulted in 0.85 of the area under the ROC curve with a sensitivity of 73% and specificity of 100% (p=0.0001). The presence of one or more of the CT findings had a sensitivity of 92%, specificity of 85% and diagnostic accuracy of 89%. Change in the ACD is a helpful CT finding for detecting open-globe injury. A difference in the ACDs {>=}0.4 mm is a helpful diagnostic criterion with excellent specificity. (orig.)

  2. One-step preparation and application of mussel-inspired poly(norepinephrine)-coated polydimethylsiloxane microchip for separation of chiral compounds.

    Science.gov (United States)

    Chen, Juan; Liang, Ru-Ping; Wu, Lu-Lu; Qiu, Jian-Ding

    2016-07-01

    In this paper, using the self-polymerization of norepinephrine (NE) and its favorable film-forming property, a simple and green preparation approach was developed to modify a PDMS channel for enantioseparation of chiral compounds. After the PDMS microchip was filled with NE solution, poly(norepinephrine) (PNE) film was gradually formed and deposited on the inner wall of microchannel as permanent coating via the oxidation of NE by the oxygen dissolved in the solution. Due to possessing plentiful catechol and amine functional groups, the PNE-coated PDMS microchip exhibited much better wettability, more stable and suppressed EOF, and less nonspecific adsorption. The water contact angle and EOF of PNE-coated PDMS substrate were measured to be 13° and 1.68 × 10(-4) cm(2) V(-1) s(-1) , compared to those of 108° and 2.24 × 10(-4) cm(2) V(-1) s(-1) from the untreated one, respectively. Different kinds of chiral compounds, such as amino acid enantiomer, drug enantiomer, and peptide enantiomer were efficiently separated utilizing a separation length of 37 mm coupled with in-column amperometric detection on the PNE-coated PDMS microchips. This facile mussel-inspired PNE-based microchip system exhibited strong recognition ability, high-performance, admirable reproducibility, and stability, which may have potential use in the complex biological analysis. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Diagnostic Value of Senior Dental Students in Yazd About of Detection the Proximal Caries on Panoramic Radiographs Compared to Detection of Experts in 1394

    Directory of Open Access Journals (Sweden)

    E Romoozi

    2016-08-01

    Full Text Available Introduction: Tooth decay is the most common chronic disease of man in the world and dentists should receive the capability to accurately diagnose of tooth decay during the training courses. In addition to clinical examination, the panoramic view and intraoral radiography is usually used for the caries detection. Therefore, the detection of caries on X-ray images can have a role in treatment planning. Methods: In this analytical study, 10 panoramic radiographies that randomly selected, separately given to 30 senior dental students and 2 professors (in order to determine the gold standard. Data were analyzed using SPSS 17 software, diagnostic tables and indexes were prepared and the results were analyzed by Kappa test. Moreover, in order to determine the agreement between the professors and students about the depth of the decay the weighted kappa coefficient was used. Results: The kappa value about detection of presence or absence of proximal caries between professors and students's diagnosis was 0.428 (P value=0.001. Diagnostic sensitivity, specificity, positive predictive value and negative predictive value obtained by students in caries detection were %47, %91.9, %63 and %85.3, respectively. Coefficient of agreement in detection of depth diagnosis obtained by professors and students was 0.361(p value=0.000. Conclusion: The diagnostic capability of senior dental students about caries detection was fair and depth diagnosis was slight.

  4. Diagnostic Efficacy of Vessel Specific Coronary Calcium Score in Detection of Coronary Artery Stenosis

    Science.gov (United States)

    Motevalli, Marzieh; Ghanaati, Hossein; Firouznia, Kavous; Kargar, Jalal; Aliyari Ghasabeh, Mounes; Shahriari, Mona; Jalali, Amir Hosein; Shakiba, Madjid

    2014-01-01

    Background: Coronary artery calcification which is determined quantitatively by coronary calcium scoring has been known as a sign of coronary stenosis and thus future cardiac events; hence it has been noticed on spotlight of researchers in recent years. Developing different method for early and optimal detection of coronary artery disease (CAD) is really essential as CAD are the first cause of death in population. Objectives: To evaluate predictive value of vessel specific coronary artery calcium (CAC) score in predicting obstructive coronary artery disease. Patients and Methods: In this diagnostic test study we evaluated patients with coronary computed tomography angiography (CCTA) and CAC score which had been referred to two referral radiology center in Tehran, Iran and finally we selected 2525 patients in a single and sequential pattern to create a diagnostic study. The whole-heart CAC scores and vessel specific CAC scores were calculated individually for the 4 major epicardial coronary arteries in 2 distinct group; group A ( patients with previous history of CABG) and group B (patients without history of CABG). For evaluation of obstruction tree cut off points were described: 0 > ; at least 1 segment with any kind of stenosis, ≥ 50; at least 1 segment with stenosis ≥ 50, ≥ 70; at least 1 segment with stenosis ≥ 70. Results: Mean of coronary calcium scores in terms of each coronary artery vessel increase by increasing coronary stenosis grade in group B; LAD, RCA, LCX respectively have mean CAC score 6.06, 6.21 and 5.04 in normal patients and 221.6, 226.7 and 106.6 in patients with complete stenosis. As expected these findings don't work for group A. Also By increasing calcium score cutoff in all four vessels sensitivity decreased and specificity increased but steal LAD had higher sensitivity than other vessels and LM had higher specificity. Thus using calcium score method is useful for ruling out stenosis in LAD while calcium score of LM can predict

  5. The diagnostic value of IADL evaluation in the detection of dementia in general practice.

    Science.gov (United States)

    De Lepeleire, J; Aertgeerts, B; Umbach, I; Pattyn, P; Tamsin, F; Nestor, L; Krekelbergh, F

    2004-01-01

    It is assumed that general practitioners can make an important contribution to the diagnosis of dementia. One of the used strategies comprises an evaluation of the Instrumental Activities of Daily Living (IADL). There are contradictory data on the value of this strategy. During one month, 21 Flemish general practitioners evaluated the IADL capacities of all subjects older than 65 years with whom they had contact. Subjects with dementia and/or living in a residential home for the elderly were excluded. Housing and living conditions, medication use and IADL were registered. The general practitioner formulated a clinical evaluation. All subjects with an IADL score > or = 1 and a random sample from the group IADL = 0 underwent a Mini Mental State Examination. Subjects with an IADL score = 4 were referred for neuropsychological and specialist examination. The average age of the 1003 registered subjects was 75.1 years (SD = 6.8). A large majority of them (85%) were totally independent. There was a large discrepancy between the family's and the patient's judgment on the presence of memory problems. There was an inverse correlation between the IADL and MMSE: when the IADL score increased, the MMSE score fell. The diagnostic value of the IADL for the diagnosis of dementia with Camdex-N as a reference standard could not be evaluated because the number of tested subjects was too small. Against the MMSE, sensitivity was 0.81 (SE = 0.03), and specificity was 0.48 (SE = 0.05). The evaluation of the IADL activities had some drawbacks as a detection method for dementia but the use of IADL data may still be clinically valuable in general practice. The correlation between the general practitioner's judgment and that of the specialist was very good. This study showed that the use of the IADL score might change the general practitioner's diagnostic judgment. Furthermore this study confirms the existence of a major threshold for the referral to a specialist of patients with suspected

  6. GryphSens: A Smartphone-Based Portable Diagnostic Reader for the Rapid Detection of Progesterone in Milk.

    Science.gov (United States)

    Jang, Hyunwook; Ahmed, Syed Rahin; Neethirajan, Suresh

    2017-05-10

    Enzyme-linked immunosorbent assay (ELISA) is a popular assay technique for the detection and quantification of various biological substances due its high sensitivity and specificity. More often, it requires large and expensive laboratory instruments, which makes it difficult to conduct when the tests must be performed quickly at the point-of-care (POC). To increase portability and ease of use, we propose a portable diagnostic system based on a Raspberry Pi imaging sensor for the rapid detection of progesterone in milk samples. We designed, assembled, and tested a standalone portable diagnostic reader and validated it for progesterone detection against a standard ELISA assay using a commercial plate reader. The portable POC device yielded consistent results, regardless of differences in the cameras and flashlights between various smartphone devices. An Android application was built to provide front-end access to users, control the diagnostic reader, and display and store the progesterone measurement on the smartphone. The diagnostic reader takes images of the samples, reads the pixel values, processes the results, and presents the results on the handheld device. The proposed POC reader can perform to superior levels of performance as a plate reader, while adding the desirable qualities of portability and ease of use.

  7. Temporal diagnostic analysis of the SWAT model to detect dominant periods of poor model performance

    Science.gov (United States)

    Guse, Björn; Reusser, Dominik E.; Fohrer, Nicola

    2013-04-01

    Hydrological models generally include thresholds and non-linearities, such as snow-rain-temperature thresholds, non-linear reservoirs, infiltration thresholds and the like. When relating observed variables to modelling results, formal methods often calculate performance metrics over long periods, reporting model performance with only few numbers. Such approaches are not well suited to compare dominating processes between reality and model and to better understand when thresholds and non-linearities are driving model results. We present a combination of two temporally resolved model diagnostic tools to answer when a model is performing (not so) well and what the dominant processes are during these periods. We look at the temporal dynamics of parameter sensitivities and model performance to answer this question. For this, the eco-hydrological SWAT model is applied in the Treene lowland catchment in Northern Germany. As a first step, temporal dynamics of parameter sensitivities are analyzed using the Fourier Amplitude Sensitivity test (FAST). The sensitivities of the eight model parameters investigated show strong temporal variations. High sensitivities were detected for two groundwater (GW_DELAY, ALPHA_BF) and one evaporation parameters (ESCO) most of the time. The periods of high parameter sensitivity can be related to different phases of the hydrograph with dominances of the groundwater parameters in the recession phases and of ESCO in baseflow and resaturation periods. Surface runoff parameters show high parameter sensitivities in phases of a precipitation event in combination with high soil water contents. The dominant parameters give indication for the controlling processes during a given period for the hydrological catchment. The second step included the temporal analysis of model performance. For each time step, model performance was characterized with a "finger print" consisting of a large set of performance measures. These finger prints were clustered into

  8. Rapid and sensitive detection of Yersinia pestis using amplification of plague diagnostic bacteriophages monitored by real-time PCR.

    Directory of Open Access Journals (Sweden)

    Kirill V Sergueev

    Full Text Available BACKGROUND: Yersinia pestis, the agent of plague, has caused many millions of human deaths and still poses a serious threat to global public health. Timely and reliable detection of such a dangerous pathogen is of critical importance. Lysis by specific bacteriophages remains an essential method of Y. pestis detection and plague diagnostics. METHODOLOGY/PRINCIPAL FINDINGS: The objective of this work was to develop an alternative to conventional phage lysis tests--a rapid and highly sensitive method of indirect detection of live Y. pestis cells based on quantitative real-time PCR (qPCR monitoring of amplification of reporter Y. pestis-specific bacteriophages. Plague diagnostic phages phiA1122 and L-413C were shown to be highly effective diagnostic tools for the detection and identification of Y. pestis by using qPCR with primers specific for phage DNA. The template DNA extraction step that usually precedes qPCR was omitted. phiA1122-specific qPCR enabled the detection of an initial bacterial concentration of 10(3 CFU/ml (equivalent to as few as one Y. pestis cell per 1-microl sample in four hours. L-413C-mediated detection of Y. pestis was less sensitive (up to 100 bacteria per sample but more specific, and thus we propose parallel qPCR for the two phages as a rapid and reliable method of Y. pestis identification. Importantly, phiA1122 propagated in simulated clinical blood specimens containing EDTA and its titer rise was detected by both a standard plating test and qPCR. CONCLUSIONS/SIGNIFICANCE: Thus, we developed a novel assay for detection and identification of Y. pestis using amplification of specific phages monitored by qPCR. The method is simple, rapid, highly sensitive, and specific and allows the detection of only live bacteria.

  9. Microchip Capillary Electrophoresis with an End-Channel Amperometric Detector and Its Preliminary Application

    Institute of Scientific and Technical Information of China (English)

    吴友谊; 屈锋; 林金明

    2005-01-01

    An end-channel amperometric detector with a guide tube for working electrode was designed and integrated on a home-made glass microchip. The guide tube was directly patterned and fabricated at the end of the detection reservoir, which made the fixation and alignment of working electrode relatively easy. The fabrication was carried out in a two-step etching process. A 30 μm carbon fiber microdisk electrode and Pt cathode were also integrated onto the amperometric detector. The baseline separation of dopamine (DA), catechol (CA) and epinephrine (EP) was achieved within 80 s. Relative standard deviations of not more than 5.2% were obtained for both peak currents and migration times of DA and CA (n=5). Using standard adding method, DA in tLrine and plasma samples was detected. The recoveries were in the range of 83%—103%.

  10. A Compact Microchip-Based Atomic Clock Based on Ultracold Trapped Rb Atoms

    CERN Document Server

    Farkas, Daniel M; Anderson, Dana Z

    2009-01-01

    We propose a compact atomic clock based on ultracold Rb atoms that are magnetically trapped near the surface of an atom microchip. An interrogation scheme that combines electromagnetically-induced transparency (EIT) with Ramsey's method of separated oscillatory fields can achieve atomic shot-noise level performance of 10^{-13}/sqrt(tau) for 10^6 atoms. The EIT signal can be detected with a heterodyne technique that provides noiseless gain; with this technique the optical phase shift of a 100 pW probe beam can be detected at the photon shot-noise level. Numerical calculations of the density matrix equations are used to identify realistic operating parameters at which AC Stark shifts are eliminated. By considering fluctuations in these parameters, we estimate that AC Stark shifts can be canceled to a level better than 2*10^{-14}. An overview of the apparatus is presented with estimates of duty cycle and power consumption.

  11. Experimental comparison of autodyne and heterodyne laser interferometry using a Nd:YVO4 microchip laser

    CERN Document Server

    Jacquin, Olivier; Glastre, Wilfried; Hugon, Olivier; De Chatellus, Hugues Guillet

    2012-01-01

    Using a Nd:YVO4 microchip laser with a relaxation frequency in the megahertz range, we have experimentally compared a heterodyne interferometer based on a Michelson configuration with an autodyne interferometer based on the laser optical feedback imaging (LOFI) method regarding their signal to noise ratios. In the heterodyne configuration, the beating between the reference beam and the signal beam is realized outside the laser cavity while in the autodyne configuration, the wave beating takes place inside the laser cavity and the relaxation oscillations of the laser intensity then play an important part. For a given laser output power, object under investigation and detection noise level, we have determined the amplification gain of the LOFI interferometer compared to the heterodyne interferometer. LOFI interferometry is demonstrated to show higher performances than heterodyne interferometry for a wide range of laser power and detection level of noise. The experimental results are in good agreement with the t...

  12. Diagnostic accuracy of a novel software technology for detecting pneumothorax in a porcine model.

    Science.gov (United States)

    Summers, Shane M; Chin, Eric J; April, Michael D; Grisell, Ronald D; Lospinoso, Joshua A; Kheirabadi, Bijan S; Salinas, Jose; Blackbourne, Lorne H

    2017-09-01

    Our objective was to measure the diagnostic accuracy of a novel software technology to detect pneumothorax on Brightness (B) mode and Motion (M) mode ultrasonography. Ultrasonography fellowship-trained emergency physicians performed thoracic ultrasonography at baseline and after surgically creating a pneumothorax in eight intubated, spontaneously breathing porcine subjects. Prior to pneumothorax induction, we captured sagittal M-mode still images and B-mode videos of each intercostal space with a linear array transducer at 4cm of depth. After collection of baseline images, we placed a chest tube, injected air into the pleural space in 250mL increments, and repeated the ultrasonography for pneumothorax volumes of 250mL, 500mL, 750mL, and 1000mL. We confirmed pneumothorax with intrapleural digital manometry and ultrasound by expert sonographers. We exported collected images for interpretation by the software. We treated each individual scan as a single test for interpretation by the software. Excluding indeterminate results, we collected 338M-mode images for which the software demonstrated a sensitivity of 98% (95% confidence interval [CI] 92-99%), specificity of 95% (95% CI 86-99), positive likelihood ratio (LR+) of 21.6 (95% CI 7.1-65), and negative likelihood ratio (LR-) of 0.02 (95% CI 0.008-0.046). Among 364 B-mode videos, the software demonstrated a sensitivity of 86% (95% CI 81-90%), specificity of 85% (81-91%), LR+ of 5.7 (95% CI 3.2-10.2), and LR- of 0.17 (95% CI 0.12-0.22). This novel technology has potential as a useful adjunct to diagnose pneumothorax on thoracic ultrasonography. Published by Elsevier Inc.

  13. Paradigm Diagnostics Salmonella Indicator Broth (PDX-SIB) for detection of Salmonella on selected environmental surfaces.

    Science.gov (United States)

    Olstein, Alan; Griffith, Leena; Feirtag, Joellen; Pearson, Nicole

    2013-01-01

    The Paradigm Diagnostics Salmonella Indicator Broth (PDX-SIB) is intended as a single-step selective enrichment indicator broth to be used as a simple screening test for the presence of Salmonella spp. in environmental samples. This method permits the end user to avoid multistep sample processing to identify presumptively positive samples, as exemplified by standard U.S. reference methods. PDX-SIB permits the outgrowth of Salmonella while inhibiting the growth of competitive Gram-negative and -positive microflora. Growth of Salmonella-positive cultures results in a visual color change of the medium from purple to yellow when the sample is grown at 37 +/- 1 degree C. Performance of PDX-SIB has been evaluated in five different categories: inclusivity-exclusivity, methods comparison, ruggedness, lot-to-lot variability, and shelf stability. The inclusivity panel included 100 different Salmonella serovars, 98 of which were SIB-positive during the 30 to 48 h incubation period. The exclusivity panel included 33 different non-Salmonella microorganisms, 31 of which were SIB-negative during the incubation period. Methods comparison studies included four different surfaces: S. Newport on plastic, S. Anatum on sealed concrete, S. Abaetetuba on ceramic tile, and S. Typhimurium in the presence of 1 log excess of Citrobacter freundii. Results of the methods comparison studies demonstrated no statistical difference between the SIB method and the U.S. Food and Drug Administration-Bacteriological Analytical Manual reference method, as measured by the Mantel-Haenszel Chi-square test. Ruggedness studies demonstrated little variation in test results when SIB incubation temperatures were varied over a 34-40 degrees C range. Lot-to-lot consistency results suggest no detectable differences in manufactured goods using two reference Salmonella serovars and one non-Salmonella microorganism.

  14. Development and validation of a novel molecular biomarker diagnostic test for the early detection of sepsis.

    Science.gov (United States)

    Sutherland, Allison; Thomas, Mervyn; Brandon, Roslyn A; Brandon, Richard B; Lipman, Jeffrey; Tang, Benjamin; McLean, Anthony; Pascoe, Ranald; Price, Gareth; Nguyen, Thu; Stone, Glenn; Venter, Deon

    2011-06-20

    Sepsis is a complex immunological response to infection characterized by early hyper-inflammation followed by severe and protracted immunosuppression, suggesting that a multi-marker approach has the greatest clinical utility for early detection, within a clinical environment focused on Systemic Inflammatory Response Syndrome (SIRS) differentiation. Pre-clinical research using an equine sepsis model identified a panel of gene expression biomarkers that define the early aberrant immune activation. Thus, the primary objective was to apply these gene expression biomarkers to distinguish patients with sepsis from those who had undergone major open surgery and had clinical outcomes consistent with systemic inflammation due to physical trauma and wound healing. This was a multi-centre, prospective clinical trial conducted across four tertiary critical care settings in Australia. Sepsis patients were recruited if they met the 1992 Consensus Statement criteria and had clinical evidence of systemic infection based on microbiology diagnoses (n = 27). Participants in the post-surgical (PS) group were recruited pre-operatively and blood samples collected within 24 hours following surgery (n = 38). Healthy controls (HC) included hospital staff with no known concurrent illnesses (n = 20). Each participant had minimally 5 ml of PAXgene blood collected for leucocyte RNA isolation and gene expression analyses. Affymetrix array and multiplex tandem (MT)-PCR studies were conducted to evaluate transcriptional profiles in circulating white blood cells applying a set of 42 molecular markers that had been identified a priori. A LogitBoost algorithm was used to create a machine learning diagnostic rule to predict sepsis outcomes. Based on preliminary microarray analyses comparing HC and sepsis groups, a panel of 42-gene expression markers were identified that represented key innate and adaptive immune function, cell cycling, WBC differentiation, extracellular remodelling and immune

  15. Comparison between Amnisure Placental Alpha Microglobulin-1 Rapid Immunoassay and Standard Diagnostic Methods for Detection of Rupture of Membranes

    Directory of Open Access Journals (Sweden)

    Beng Kwang Ng

    2013-01-01

    Full Text Available Objective. To determine the diagnostic accuracy of placental alpha microglobulin-1 assay and standard diagnostic methods for detecting rupture of membrane. Study Design. Prospective diagnostic study, between June 2011 to November 2011 at a tertiary centre. Initial evaluation included both the standard diagnostic methods for rupture of membranes and placental alpha microglobulin-1 immunoassay. The actual rupture of membranes was diagnosed on review of the medical records after delivery (absence of membrane or a positive pad chart. Main Outcome Measures. Placental alpha microglobulin-1 immunoassay and standard diagnostic methods for diagnosis of rupture of membrane. Results. A total of 211 patients were recruited. At initial presentation, 187 patients (88.6% had ruptured membranes, while 24 patients (11.4% had intact membranes. Placental alpha microglobulin-1 immunoassay confirmed rupture of membranes at initial presentation with a sensitivity of 95.7% (179 of 187, specificity of 100% (24 of 24, positive predictive value of 100% (179 of 179, and negative predictive value of 75.0% (24 of 32. By comparison, the conventional standard diagnostic methods had a sensitivity of 78.1% (146 of 187, specificity of 100% (24 of 24, positive predictive value of 100% (146 of 146, and negative predictive value of 36.9% (24 of 65 in diagnosing rupture of membrane. Conclusion. Placental alpha-microglobulin-1 immunoassay is a rapid and accurate method for confirming the diagnosis of rupture of membrane. It was superior to conventional standard diagnostic methods (pooling, nitrazine, and ferning, the nitrazine test alone or fern test alone.

  16. Diagnostic performance of CT versus MR in detecting aldosterone-producing adenoma in primary hyperaldosteronism (Conn's syndrome)

    Energy Technology Data Exchange (ETDEWEB)

    Lingam, R.K.; Sohaib, S.A.; Rockall, A.G. [St Bartholomew' s Hospital, Department of Diagnostic Imaging, London (United Kingdom); Isidori, A.M.; Chew, S.; Monson, J.P.; Grossman, A.; Besser, G.M. [St Bartholomew' s Hospital, Department of Endocrinology, London (United Kingdom); Reznek, R.H. [St Bartholomew' s Hospital, Department of Diagnostic Imaging, London (United Kingdom); St Bartholomew' s Hospital, Academic Department of Radiology, Dominion House, London (United Kingdom)

    2004-10-01

    The aim of the present study is to compare the diagnostic performance of CT and MR imaging in detecting aldosterone-producing adenoma and to compare the interobserver variability in the detection of an aldosterone-producing adenoma on CT and MR. A retrospective study of 34 patients with primary hyperaldosteronism was performed. A total of 17 cases of aldosterone-producing adenoma and 17 cases of bilateral adrenal hyperplasia were included. The final diagnosis of an adenoma was made by surgery with histological confirmation, whereas that of bilateral adrenal hyperplasia was made on adrenal venous sampling or a good biochemical and clinical response following medical treatment alone and in the absence of a unilateral radiological abnormality. The CT (n=30) and MR (n=24) scans were reviewed independently by two radiologists experienced in adrenal imaging, who were unaware of the cause of the primary hyperaldosteronism. The diagnostic performances of both observers in detecting an aldosterone-producing adenoma on CT and MR imaging were compared. The 16 adenomatous nodules that were detected on imaging ranged from 1 to 4.75 cm in diameter. The calculated sensitivity and specificity for detecting aldosterone-producing adenoma were 87 and 93% for one observer and 85 and 82% for the other observer on CT, and 83 and 83% for one observer and 92 and 92% for the other observer on MR, respectively. Receptor operating characteristics curve analysis showed similar performances of both observers in detecting an aldosterone-producing adenoma on CT and MR imaging. There was good interobserver agreement on CT (k=0.71) and on MR (k=0.67). We have demonstrated comparable diagnostic performance and good interobserver agreement on CT and MR imaging for the detection of aldosterone-producing adenoma. (orig.)

  17. Strengthening the diagnostic capacity to detect Bio Safety Level 3 organisms in unusual respiratory viral outbreaks.

    NARCIS (Netherlands)

    Asten, L. van; Lubben, M. van der; Wijngaard, C. van den; Pelt, W. van; Verheij, R.; Jacobi, A.; Overduin, P.; Meijer, A.; Luijt, D.; Claas, E.; Hermans, M.; Melchers, W.J.G.; Rossen, J.; Schuurman, R.; Wolffs, P.; Bouchier, C.; Schirm, J.; Kroes, L.; Leenders, S.; Galama, J.M.D.; Peeters, M.; Loon, A. van; Stobberingh, E.E.; Schutten, M.; Koopmans, M.

    2009-01-01

    BACKGROUND: Experience with a highly pathogenic avian influenza outbreak in the Netherlands (2003) illustrated that the diagnostic demand for respiratory viruses at different biosafety levels (including BSL3), can increase unexpectedly and dramatically. OBJECTIVES: We describe the measures taken sin

  18. Strengthening the diagnostic capacity to detect Bio Safety Level 3 organisms in unusual respiratory viral outbreaks

    NARCIS (Netherlands)

    van Asten, Liselotte; van der Lubben, Mariken; van den Wijngaard, Cees; van Pelt, Wilfrid; Verheij, Robert; Jacobi, Andre; Overduin, Pieter; Meijer, Adam; Luijt, Dirk; Claas, Eric; Hermans, Mirjam; Melchers, Willem; Rossen, John; Schuurman, Rob; Wolffs, Petra; Bouchier, Charles; Schirm, Jurjen; Kroes, Louis; Leenders, Sander; Galama, Joep; Peeters, Marcel; van Loon, Anton; Stobberingh, Ellen; Schutten, Martin; Koopmans, Marion D. V. M.

    2009-01-01

    Background: Experience with a highly pathogenic avian influenza outbreak in the Netherlands (2003) illustrated that the diagnostic demand for respiratory viruses at different biosafety levels (including BSL3), can increase unexpectedly and dramatically. Objectives: We describe the measures taken sin

  19. Diagnostic methods to cutaneous leishmaniasis detection in domestic dogs and cats

    National Research Council Canada - National Science Library

    Trevisan, Daliah Alves Coelho; Lonardoni, Maria Valdrinez Campana; Demarchi, Izabel Galhardo

    2015-01-01

    Cutaneous leishmaniasis is caused by different species of Leishmania. In domestic animals such as dogs and cats, the diagnostic consists of clinical, epidemiological and serological tests, which changes among countries all around the world...

  20. Microchip electrophoresis-copper nanowires for fast and reliable determination of monossacharides in honey samples.

    Science.gov (United States)

    García, Miguel; Escarpa, Alberto

    2014-02-01

    Microchip electrophoresis (ME) with electrochemical detection has been demonstrated to be a powerful tool in food analysis. However, the coupling of ME with electrochemical detection and nanotechnologies is still in its infancy, knowing that nanomaterials can significantly improve the ME analytical performance. This work reports the coupling between ME and copper nanowires (CuNWs) for the selective analysis of monosaccharides in honey samples. Also, in terms of real applicability, the study of analytical reliability of ME is an issue of paramount importance. To this end, a representative group of nine honey samples were analyzed and the results were compared with those previously obtained by HPLC-refractive index. ME-CuNWs approach allowed the separation of glucose and fructose in <250 s under optimized separation (20 mM NaOH + 10 mM H3 BO3 , pH 12; separation voltage + 1000 V) and detection (E = +0.70 V in 20 mM NaOH + 10 mM H3 BO3 , pH 12) conditions. An excellent stability of EOF during sample analysis was achieved with RSDs for migration times <2% and for amperometric currents <9%. The quantitative contents for individual glucose and fructose obtained using ME-CuNWs in comparison with those obtained by HPLC-refractive index were highly in agreement with errors <10% indicating the reliability of the approach. The excellent analytical performance obtained confirms the analytical potency of ME-CuNWs approach, enhancing the maturity of the microchip technology and opening new avenues for future implementation of applications in the field of food analysis.

  1. Selective manipulation of superparamagnetic beads by a magnetic microchip

    KAUST Repository

    Gooneratne, Chinthaka Pasan

    2013-07-01

    In this paper, a magnetic microchip (MMC) is presented, to first trap and then selectively manipulate individual, superparamagnetic beads (SPBs) to another trapping site. Trapping sites are realized through soft magnetic micro disks made of Ni80Fe20, and SPB motion is controlled by current-carrying, tapered, conducting lines made of Au. The MMC was realized using standard microfabrication techniques and provides a cheap and versatile platform for microfluidic systems for cell manipulation. © 2013 IEEE.

  2. Simultaneous immunoassay analysis of plasma IL-6 and TNF-α on a microchip.

    Directory of Open Access Journals (Sweden)

    Kaori Abe

    Full Text Available Sandwich enzyme-linked immunosorbant assay (ELISA using a 96-well plate is frequently employed for clinical diagnosis, but is time-and sample-consuming. To overcome these drawbacks, we performed a sandwich ELISA on a microchip. The microchip was made of cyclic olefin copolymer with 4 straight microchannels. For the construction of the sandwich ELISA for interleukin-6 (IL-6 or tumor necrosis factor-α (TNF-α, we used a piezoelectric inkjet printing system for the deposition and fixation of the 1st anti-IL-6 antibody or 1st anti-TNF-α antibody on the surface of the each microchannel. After the infusion of 2 µl of sample to the microchannel and a 20 min incubation, 2 µl of biotinylated 2nd antibody for either antigen was infused and a 10 min incubation. Then 2 µl of avidin-horseradish peroxidase was infused; and after a 5 min incubation, the substrate for peroxidase was infused, and the luminescence intensity was measured. Calibration curves were obtained between the concentration and luminescence intensity over the range of 0 to 32 pg/ml (IL-6: R(2 = 0.9994, TNF-α: R(2 = 0.9977, and the detection limit for each protein was 0.28 pg/ml and 0.46 pg/ml, respectively. Blood IL-6 and TNF-α concentrations of 5 subjects estimated from the microchip data were compared with results obtained by the conventional method, good correlations were observed between the methods according to linear regression analysis (IL-6: R(2 = 0.9954, TNF-α: R(2 = 0.9928. The reproducibility of the presented assay for the determination of the blood IL-6 and TNF-α concentration was comparable to that obtained with the 96-well plate. Simultaneous detection of blood IL-6 and TNF-α was possible by the deposition and fixation of each 1st antibody on the surface of a separate microchannel. This assay enabled us to determine simultaneously blood IL-6 and TNF-α with accuracy, satisfactory sensitivity, time saving ability, and low consumption of sample and

  3. Atmospheric pressure thermospray ionization using a heated microchip nebulizer.

    Science.gov (United States)

    Keski-Rahkonen, Pekka; Haapala, Markus; Saarela, Ville; Franssila, Sami; Kotiaho, Tapio; Kostiainen, Risto; Auriola, Seppo

    2009-10-30

    When a standard atmospheric pressure chemical ionization (APCI) or atmospheric pressure photoionization (APPI) ion source is used without applying the corona discharge or photoirradiation, atmospheric pressure thermospray ionization (APTSI) of various compounds can be achieved. Although largely ignored, this phenomenon has recently gained interest as an alternative ionization technique. In this study, this technique is performed for the first time on a miniaturized scale using a microchip nebulizer. Sample ionization with the presented microchip-APTSI (microAPTSI) is achieved by applying only heat and gas flow to a nebulizer chip, without any other methods to promote gas-phase ionization. To evaluate the performance of the described microAPTSI setup, ionization efficiency for a set of test compounds was monitored as the microchip positioning, temperature, nebulizer gas flow rate, sample solution composition, and solvent flow rate were varied. The microAPTSI mass spectra of the test compounds were also compared to those obtained with ESI and APCI. The microAPTSI produces ESI-like spectra with low background noise, favoring the formation of protonated or deprotonated molecules of compounds that are ionizable in solution. Multiple charging of peptides without in-source fragmentation was also observed. Unlike ESI, however, the microAPTSI source can tolerate the presence of mobile phase additives like trifluoroacetic acid (TFA) without significant ion suppression. The microAPTSI source can be used with standard mass spectrometer ion source hardware, being a unique alternative to the present interfacing techniques.

  4. Disposable polyester-toner electrophoresis microchips for DNA analysis.

    Science.gov (United States)

    Duarte, Gabriela R M; Coltro, Wendell K T; Borba, Juliane C; Price, Carol W; Landers, James P; Carrilho, Emanuel

    2012-06-07

    Microchip electrophoresis has become a powerful tool for DNA separation, offering all of the advantages typically associated with miniaturized techniques: high speed, high resolution, ease of automation, and great versatility for both routine and research applications. Various substrate materials have been used to produce microchips for DNA separations, including conventional (glass, silicon, and quartz) and alternative (polymers) platforms. In this study, we perform DNA separation in a simple and low-cost polyester-toner (PeT)-based electrophoresis microchip. PeT devices were fabricated by a direct-printing process using a 600 dpi-resolution laser printer. DNA separations were performed on PeT chip with channels filled with polymer solutions (0.5% m/v hydroxyethylcellulose or hydroxypropylcellulose) at electric fields ranging from 100 to 300 V cm(-1). Separation of DNA fragments between 100 and 1000 bp, with good correlation of the size of DNA fragments and mobility, was achieved in this system. Although the mobility increased with increasing electric field, separations showed the same profile regardless of the electric field. The system provided good separation efficiency (215,000 plates per m for the 500 bp fragment) and the separation was completed in 4 min for 1000 bp fragment ladder. The cost of a given chip is approximately $0.15 and it takes less than 10 minutes to prepare a single device.

  5. Hydrodynamic injection on electrophoresis microchips using an electronic micropipette.

    Science.gov (United States)

    Gabriel, Ellen F M; Dos Santos, Rodrigo A; Lobo-Júnior, Eulício O; Rezende, Kariolanda C A; Coltro, Wendell K T

    2017-01-01

    Here we report for the first time the use of an electronic micropipette as hydrodynamic (HD) injector for microchip electrophoresis (ME) devices. The micropipette was directly coupled to a PDMS device, which had been fabricated in a simple cross format with two auxiliary channels for sample volume splitting. Sample flow during the injection procedure was controlled in automatic dispenser mode using a volume of 0.6µL. Channel width and device configuration were optimized and the best results were achieved using a simple cross layout containing two auxiliary channels with 300µm width for sample splitting. The performance of the HD injector was evaluated using a model mixture of high-mobility cationic species. The results obtained were compared to the data obtained via electrokinetic (EK) injection. Overall, the HD provided better analytical performance in terms of resolution and injection-to-injection repeatability. The relative standard deviation (RSD) values for peak intensities were lower than 5% (n=10) when the micropipette was employed. In comparison with EK injection, the use of the proposed HD injector revealed an unbiased profile for a mixture containing K(+) and Li(+)(300 µmol L(-1) each) over various buffer concentrations. For EK injection, the peak areas decreased from 2.92 ± 0.20-0.72 ± 0.14Vs for K(+) and from 1.30 ± 0.10-0.38 ± 0.10Vs for Li(+) when the running buffer increased from 20 to 50mmolL(-1). For HD injection, the peak areas for K(+) and Li(+) exhibited average values of 2.48±0.07 and 2.10±0.06Vs, respectively. The limits of detection (LDs) for K(+), Na(+) and Li(+) ranged from 18 to 23µmolL(-1). HD injection through an electronic micropipette allows to automatically dispense a bias-free amount of sample inside microchannels with acceptable repeatability. The proposed approach also exhibited instrumental simplicity, portability and minimal microfabrication requirements.

  6. A fully integrated paperfluidic molecular diagnostic chip for the extraction, amplification, and detection of nucleic acids from clinical samples.

    Science.gov (United States)

    Rodriguez, Natalia M; Wong, Winnie S; Liu, Lena; Dewar, Rajan; Klapperich, Catherine M

    2016-02-21

    Paper diagnostics have successfully been employed to detect the presence of antigens or small molecules in clinical samples through immunoassays; however, the detection of many disease targets relies on the much higher sensitivity and specificity achieved via nucleic acid amplification tests (NAAT). The steps involved in NAAT have recently begun to be explored in paper matrices, and our group, among others, has reported on paper-based extraction, amplification, and detection of DNA and RNA targets. Here, we integrate these paper-based NAAT steps into a single paperfluidic chip in a modular, foldable system that allows for fully integrated fluidic handling from sample to result. We showcase the functionality of the chip by combining nucleic acid isolation, isothermal amplification, and lateral flow detection of human papillomavirus (HPV) 16 DNA directly from crude cervical specimens in less than 1 hour for rapid, early detection of cervical cancer. The chip is made entirely of paper and adhesive sheets, making it low-cost, portable, and disposable, and offering the potential for a point-of-care molecular diagnostic platform even in remote and resource-limited settings.

  7. Silica-Based Solid Phase Extraction of DNA on a Microchip

    Institute of Scientific and Technical Information of China (English)

    陈晓芳; 沈科跃; 刘鹏; 郭旻; 程京; 周玉祥

    2004-01-01

    Micro total analysis systems for chemical and biological analysis have attracted much attention.However,microchips for sample preparation and especially DNA purification are still underdeveloped.This work describes a solid phase extraction chip for purifying DNA from biological samples based on the adsorption of DNA on bare silica beads prepacked in a microchannel.The chip was fabricated with poly-dimethylsiloxane.The silica beads were packed in the channel on the chip with a tapered microchannel to form the packed bed.Fluorescence detection was used to evaluate the DNA adsorbing efficiency of the solid phase.The polymerase chain reaction was used to evaluate the quality of the purified DNA for further use.The extraction efficiency for the DNA extraction chip is approximately 50% with a 150-nL extraction volume.Successful amplification of DNA extracted from human whole blood indicates that this method is compatible with the polymerase chain reaction.

  8. Dual-focus Magnification, High-Definition Endoscopy Improves Pathology Detection in Direct-to-Test Diagnostic Upper Gastrointestinal Endoscopy.

    Science.gov (United States)

    Bond, Ashley; Burkitt, Michael D; Cox, Trevor; Smart, Howard L; Probert, Chris; Haslam, Neil; Sarkar, Sanchoy

    2017-03-01

    In the UK, the majority of diagnostic upper gastrointestinal (UGI) endoscopies are a result of direct-to-test referral from the primary care physician. The diagnostic yield of these tests is relatively low, and the burden high on endoscopy services. Dual-focus magnification, high-definition endoscopy is expected to improve detection and classification of UGI mucosal lesions and also help minimize biopsies by allowing better targeting. This is a retrospective study of patients attending for direct-to-test UGI endoscopy from January 2015 to June 2015. The primary outcome of interest was the identification of significant pathology. Detection of significant pathology was modelled using logistic regression. 500 procedures were included. The mean age of patients was 61.5 (±15.6) years; 60.8% of patients were female. Ninety-four gastroscopies were performed using dual-focus magnification high-definition endoscopy. Increasing age, male gender, type of endoscope, and type of operator were all identified as significant factors influencing the odds of detecting significant mucosal pathology. Use of dual-focus magnification, high-definition endoscopy was associated with an odds ratio of 1.87 (95%CI 1.11-3.12) favouring the detection of significant pathology. Subsequent analysis suggested that the increased detection of pathology during dual-focus magnification, high-definition endoscopy also influenced patient follow-up and led to a 3.0 fold (p=0.04) increase in the proportion of patients entered into an UGI endoscopic surveillance program. Dual-focus magnification, high-definition endoscopy improved the diagnostic yield for significant mucosal pathology in patients referred for direct-to-test endoscopy. If this finding is recapitulated elsewhere it will have substantial impact on the provision of UGI endoscopic services.

  9. Diagnostic use of PCR for detection of Pneumocystis carinii in oral wash samples

    DEFF Research Database (Denmark)

    Helweg-Larsen, J; Jensen, Jens Ulrik Stæhr; Benfield, T;

    1998-01-01

    and was compared to a previously described PCR protocol (mitochondrial RNA) run in a research laboratory. Both PCR methods amplified a sequence of the mitochondrial rRNA gene of P. carinii. Paired bronchoalveolar lavage (BAL) and oral wash specimens from 76 consecutive human immunodeficiency virus type 1-infected...... specimens and 100, 91, 90, and 100%, respectively, for BAL specimens. Our results suggest that oral wash specimens are a potential noninvasive method to obtain a diagnostic specimen during P. carinii pneumonia infection and that it can be applied in a routine diagnostic laboratory....

  10. A new rapid diagnostic test for detection of anti-Schistosoma mansoni and anti-Schistosoma haematobium antibodies

    Directory of Open Access Journals (Sweden)

    Coulibaly Jean T

    2013-01-01

    Full Text Available Abstract Background Parasitological methods are widely used for the diagnosis of schistosomiasis. However, they are insensitive, particularly in areas of low endemicity, and labour-intensive. Immunoassays based on detection of anti-schistosome antibodies have the merit of high sensitivity and recently a rapid diagnostic test (RDT, incorporating Schistosoma mansoni cercarial transformation fluid (SmCTF for detection of anti-schistosome antibodies in blood has been developed. Here, we assessed the diagnostic performance of the SmCTF-RDT for S. mansoni and S. haematobium infections by comparing it with microscopy for egg detection. Methods A cross-sectional survey was carried out in Azaguié, south Côte d’Ivoire. 118 pre-school-aged children submitted two stool and two urine samples, which were subjected to the Kato-Katz and urine filtration methods for the detection of S. mansoni and S. haematobium eggs, respectively. Urine was also subjected to a commercially available cassette test for S. mansoni, which detects circulating cathodic antigen. A finger-prick blood sample was used for the SmCTF-RDT for detection of anti-S. mansoni and anti-S. haematobium antibodies. Results The prevalence of both anti-S. mansoni and anti-S. haematobium antibodies was more than three times higher than the prevalence of infection estimated by egg detection under a microscope. Using quadruplicate Kato-Katz as the reference standard for the diagnosis of S. mansoni infection, the sensitivity, negative predictive value (NPV, and positive predictive value (PPV of the SmCTF-RDT was 75.0%, 84.2% and 22.5%, respectively. When two urine filtrations were considered as the reference standard for the diagnosis of S. haematobium infection, the sensitivity, NPV and PPV of SmCTF-RDT was 66.7%, 94.9% and 5.1%, respectively. The specificity of SmCTF-RDT, when using egg-detection as the reference standard, was estimated to be 34.4%. This low specificity may be a reflection of the

  11. Methylene Blue as a Diagnostic Aid in the Early Detection of Potentially Malignant and Malignant Lesions of Oral Mucosa.

    Science.gov (United States)

    Lejoy, Abraham; Arpita, Rai; Krishna, Burde; Venkatesh, Naikmasur

    2016-05-01

    In vivo stains are the prompt resources, which have emerged in recent years to aid as clinical diagnostic tools in detecting early potentially malignant and malignant lesions. Toluidine blue, by its property of retaining in the increased DNA and RNA cellular activity areas, aids in delineating the suspicious areas. However, it is hazardous if swallowed, and has been shown to have toxicity to fibroblasts. Methylene blue has a similar chemical structure and exhibits similar physicochemical properties as toluidine blue. It is less toxic to the human body and has recently been proposed for screening some gastrointestinal or prostate tumors. The application of this material in detecting oral lesions has so far not been addressed. The objective of this study was to evaluate the sensitivity and reliability of in vivo staining with methylene blue as a diagnostic adjunct in screening for oral malignant or potentially malignant lesions. The present study involved the examination of 75 patients suspected of having oral malignant or potentially malignant lesions by methylene blue staining. The results of methylene blue uptake were compared with a simultaneous biopsy of these lesions. The overall sensitivity was 95% (100% for malignancy and 92% for potentially malignant lesions) and specificity was 70%. The positive predictive value was 91% and negative predictive value of 80% was observed in the study. We consider that methylene blue staining is a useful diagnostic adjunct in a large, community-based oral cancer screening program for high-risk individuals.

  12. Ischemic Stroke Detection System with a Computer-Aided Diagnostic Ability Using an Unsupervised Feature Perception Enhancement Method

    Directory of Open Access Journals (Sweden)

    Yeu-Sheng Tyan

    2014-01-01

    Full Text Available We propose an ischemic stroke detection system with a computer-aided diagnostic ability using a four-step unsupervised feature perception enhancement method. In the first step, known as preprocessing, we use a cubic curve contrast enhancement method to enhance image contrast. In the second step, we use a series of methods to extract the brain tissue image area identified during preprocessing. To detect abnormal regions in the brain images, we propose using an unsupervised region growing algorithm to segment the brain tissue area. The brain is centered on a horizontal line and the white matter of the brain’s inner ring is split into eight regions. In the third step, we use a coinciding regional location method to find the hybrid area of locations where a stroke may have occurred in each cerebral hemisphere. Finally, we make corrections and mark the stroke area with red color. In the experiment, we tested the system on 90 computed tomography (CT images from 26 patients, and, with the assistance of two radiologists, we proved that our proposed system has computer-aided diagnostic capabilities. Our results show an increased stroke diagnosis sensitivity of 83% in comparison to 31% when radiologists use conventional diagnostic images.

  13. Ischemic stroke detection system with a computer-aided diagnostic ability using an unsupervised feature perception enhancement method.

    Science.gov (United States)

    Tyan, Yeu-Sheng; Wu, Ming-Chi; Chin, Chiun-Li; Kuo, Yu-Liang; Lee, Ming-Sian; Chang, Hao-Yan

    2014-01-01

    We propose an ischemic stroke detection system with a computer-aided diagnostic ability using a four-step unsupervised feature perception enhancement method. In the first step, known as preprocessing, we use a cubic curve contrast enhancement method to enhance image contrast. In the second step, we use a series of methods to extract the brain tissue image area identified during preprocessing. To detect abnormal regions in the brain images, we propose using an unsupervised region growing algorithm to segment the brain tissue area. The brain is centered on a horizontal line and the white matter of the brain's inner ring is split into eight regions. In the third step, we use a coinciding regional location method to find the hybrid area of locations where a stroke may have occurred in each cerebral hemisphere. Finally, we make corrections and mark the stroke area with red color. In the experiment, we tested the system on 90 computed tomography (CT) images from 26 patients, and, with the assistance of two radiologists, we proved that our proposed system has computer-aided diagnostic capabilities. Our results show an increased stroke diagnosis sensitivity of 83% in comparison to 31% when radiologists use conventional diagnostic images.

  14. Diagnostic performance of magnetic resonance venography in the detection of recanalization in patients with chronic cerebral venous sinus thrombus

    Institute of Scientific and Technical Information of China (English)

    SUN Ying; ZHENG Dong-you; JI Xun-ming; Peter WEALE; WU Hao; JIANG Li-dan; YANG Li-zhuang

    2009-01-01

    Background In the chronic stage of cerebral venous sinus thrombosis (CVST), recanalization can result in disparate MR appearances. We aimed to prospectively investigate the diagnostic accuracy of magnetic resonance venography (MRV) in the evaluation of the recanalization of CVST.Methods This study prospectively evaluated the diagnostic performance of 2-dimensional time-of-flight (2D-TOF) MRV in thirty-two consecutive patients during a three- to six-month follow-up for CVST. Both 2D-TOF MRV and digital substraction angiography (DSA) were undertaken. Diagnostic accuracy of 2D-TOF MRV in the detection of recanalized thrombus was evaluated using DSA as the reference standard.Results MRV and DSA were completed without complications in all 32 patients. The sensitivity, specificity, positive predictive value, and negative predictive value of 2D-TOF MRV for the detection of recanalization on a segmental basis were 91% (62/68), 93% (37/40), 95% (62/65), and 86% (37/43) respectively.Conclusion 2D-TOF MRV provides high sensitivity and specificity for the diagnosis of recanalized CVST segments.

  15. Urine steroid metabolomics as a novel diagnostic tool for early detection of recurrence in adrenocortical carcinoma

    NARCIS (Netherlands)

    Chortis, Vasileios; Bancos, Irina; Lang, Katharina; Hughes, Beverly A.; O'Neil, Donna M.; Taylor, Angela E.; Fassnacht, Martin; Bertherat, Jerome; Beuschlein, Felix; Quinkler, Marcus; Vassiliadi, Dimitri; Conall Dennedy, M; Mannelli, Massimo; Biehl, Michael; Arlt, Wiebke

    2015-01-01

    Introduction: Adrenocortical carcinoma (ACC) is an aggressive malignancy with a high rate of recurrence. Regular post-operative follow-up imaging is necessary, but associated with high radiation exposure and frequent diagnostic ambiguity. Urine steroid metabolomics has recently been introduced as a

  16. Diagnostic accuracy of nocturnal oximetry for detection of sleep apnea syndrome in stroke rehabilitation

    NARCIS (Netherlands)

    J.A. Aaronson; T. van Bezeij; J.G. van den Aardweg; C.A.M. van Bennekom; W.F. Hofman

    2012-01-01

    Background and Purpose—Sleep apnea syndrome (SAS) is a common sleep disorder in stroke patients and is associated with decreased recovery and increased risk of recurrent stroke and mortality. The standard diagnostic test for SAS is poly(somno)graphy, but this is often not feasible in stroke rehabili

  17. Diagnostic accuracy of metronome-paced tachypnea to detect dynamic hyperinflation

    NARCIS (Netherlands)

    Lahaije, A.J.M.C.; Willems, L.M.; Hees, H.W. van; Dekhuijzen, P.N.R.; Helvoort, H.A.C. van; Heijdra, Y.F.

    2013-01-01

    INTRODUCTION: This prospective study was carried out to investigate if metronome-paced tachypnea (MPT) can serve as an accurate diagnostic tool to identify patients with chronic obstructive pulmonary disease (COPD) who are susceptible to develop dynamic hyperinflation during exercise. Commonly, this

  18. Diagnostic accuracy of static CT perfusion for the detection of myocardial ischemia

    DEFF Research Database (Denmark)

    Sørgaard, Mathias Holm; Kofoed, Klaus Fuglsang; Linde, Jesper James

    2016-01-01

    : Systematic literature review and meta-analysis of studies examining the diagnostic accuracy of static CTP imaging alone or combined with coronary CT angiography (CTA) in comparison to single photon emission computed tomography (SPECT), magnetic resonance perfusion (MRP), and/or invasive coronary angiography...

  19. Diagnostic utility of zinc protoporphyrin to detect iron deficiency in Kenyan preschool children

    NARCIS (Netherlands)

    Teshome, Emily M.; Prentice, Andrew M.; Demir, Ayşe Y.; Andang'o, Pauline E.A.; Verhoef, Hans

    2017-01-01

    Background: Zinc protoporphyrin (ZPP) has been used to screen and manage iron deficiency in individual children, but it has also been recommended to assess population iron status. The diagnostic utility of ZPP used in combination with haemoglobin concentration has not been evaluated in pre-school

  20. The relative diagnostic yields of clinical, FOTI and radiographic examinations for the detection of approximal caries in youngsters

    Directory of Open Access Journals (Sweden)

    Mialhe Fabio

    2009-01-01

    Full Text Available Background: The detection of carious lesions in the initial stages of development is very important in order to prevent the occurrence of cavitation. Aims and Objectives: The aim of the present study was to compare three methods-visual examination, FOTI, and bitewing radiographic examination-for the detection of noncavitated and cavitated approximal carious lesions. Materials and Methods: Seventy students (mean age 14 years with low caries prevalence were examined by three examiners. The tooth surfaces judged as having caries by at least one examiner or one diagnostic method were scheduled for tooth separation. Results: The results showed that the incorporation of FOTI and radiographic examination represented an additional diagnostic yield of 50% and about 110%, respectively, compared to clinical examination alone. Conclusion: We conclude that FOTI or radiographic examination, or both, used as adjuncts to clinical examination, could improve the detection of noncavitated and cavitated approximal carious lesions. Although FOTI should not replace bitewing radiographic examination, it does seem to have additional value for the detection of carious lesions on approximal surfaces of the posterior teeth in school children with low caries prevalence.

  1. Quantitative analysis of serum procollagen type I C-terminal propeptide by immunoassay on microchip.

    Directory of Open Access Journals (Sweden)

    Shouki Yatsushiro

    Full Text Available BACKGROUND: Sandwich enzyme-linked immunosorbent assay (ELISA is one of the most frequently employed assays for clinical diagnosis, since this enables the investigator to identify specific protein biomarkers. However, the conventional assay using a 96-well microtitration plate is time- and sample-consuming, and therefore is not suitable for rapid diagnosis. To overcome these drawbacks, we performed a sandwich ELISA on a microchip. METHODS AND FINDINGS: The microchip was made of cyclic olefin copolymer with straight microchannels that were 300 µm wide and 100 µm deep. For the construction of a sandwich ELISA for procollagen type I C-peptide (PICP, a biomarker for bone formation, we used a piezoelectric inkjet printing system for the deposition and fixation of the 1st anti-PICP antibody on the surface of the microchannel. After the infusion of the mixture of 2.0 µl of peroxidase-labeled 2nd anti-PICP antibody and 0.4 µl of sample to the microchannel and a 30-min incubation, the substrate for peroxidase was infused into the microchannel; and the luminescence intensity of each spot of 1st antibody was measured by CCD camera. A linear relationship was observed between PICP concentration and luminescence intensity over the range of 0 to 600 ng/ml (r(2 = 0.991, and the detection limit was 4.7 ng/ml. Blood PICP concentrations of 6 subjects estimated from microchip were compared with results obtained by the conventional method. Good correlation was observed between methods according to simple linear regression analysis (R(2 = 0.9914. The within-day and between-days reproducibilities were 3.2-7.4 and 4.4-6.8%, respectively. This assay reduced the time for the antigen-antibody reaction to 1/6, and the consumption of samples and reagents to 1/50 compared with the conventional method. CONCLUSION: This assay enabled us to determine serum PICP with accuracy, high sensitivity, time saving ability, and low consumption of sample and reagents, and thus

  2. Particle transport in a He-microchip plasma atomic emission system with an ultrasonic nebulizer for aqueous sample introduction

    Science.gov (United States)

    Oh, Joosuck; Lim, H. B.

    2008-11-01

    The transport efficiency of dried particles generated from an ultrasonic nebulizer (USN) was studied to improve the analytical performance of a lab-made, He-microchip plasma system, in which a quartz tube (~ 1 mm i.d.) was positioned inside the central channel of a poly(dimethylsiloxane) (PDMS) polymer chip. The polymer microchip plasma has the advantages of low cost, small size, easy handling and design, and self-ignition with long stabilization (> 24 h). However, direct introduction of aqueous solution into the microplasma for the detection of metals remains problematic due to plasma instability. In addition, the much smaller size of the system can cause signal suppression due to low transport efficiency. Therefore, knowledge of particle transport efficiency in this microplasma system is required to enhance the sensitivity and stability. The weight of transported particles in the range of 0.02 to 10 mg m - 3 was measured using a piezobalance with a precision of 0.4-17.8%, depending on the operating conditions. The significant effects of the USN operating conditions and the physical properties of the tubing, namely, length, inner diameter and surface characteristics, on the number of particles transported from the nebulizer to the microplasma were studied. When selected metals, such as Na, Mg and Pb, at a concentration of 5 mg L - 1 were nebulized, transported particles were obtained with a mass range of 0.5-5 mg m - 3 , depending on atomic weights. For application of the He-rf-microplasma, the atomic emission system was optimized by changing both the radio frequency (rf) power (60-200 W) and cooling temperature of the USN (- 12-9 °C). The limits of detection obtained for K, Na and Cu were 0.26, 0.22, and 0.28 mg L - 1 , respectively. These results confirmed the suitable stability and sensitivity of the He-rf-PDMS microchip plasma for application as an atomization source.

  3. [Diagnostic evaluation of endouterine manual aspiration for the detection of endometrial hyperplasia and cancer in patients with abnormal uterine bleeding].

    Science.gov (United States)

    Suarez Rincón, A E; Arévalo Lagunas, I; Cerpa Batres, M G; Díaz Rodríguez, M C

    2000-01-01

    The objective was to determinate the diagnostic value of manual vacuum aspiration with Karman cannula (MVA) for the detection of endometrial hyperplasia and cancer in patients with abnormal uterine bleeding. Fifty patients with abnormal uterine bleeding were evaluated with MVA prior to dilatation and curettage (D&C). The needing of cervical dilatation was noted. A matched analysis of the histological reports with Wilcoxon contrast test was performed. In order to calculate the diagnostic value, the histological examination of the tissue recollected by D&C was defined as gold-standard. Sensitivity, specificity, pre-test probability (prevalence), post-test probabilities (predictive values) and likelihood-ratios were calculated. No significant difference between either histological reports in matched analysis and the insufficient samples proportion was detected. Cervical dilatation was performed more frequently to D&C (p = 0.0002). The pre-test probability (prevalence) of endometrial hyperplasia/cancer was 20%. Two cases of hyperplasia were not detected by MVA (negative false 20%). The endometrial biopsy for MVA showed a sensitivity of 71% and specificity 93%. The post-test probabilities for an abnormal and normal biopsy (positive and negative predictive values) were 62.5% and 95.2%, respectively. The corresponding likelihood-ratios were 10.23 and 0.3, respectively. The endometrial biopsy for MVA has a high diagnostic value, similar to D&C, in the detection of endometrial hyperplasia/cancer in patients with abnormal uterine bleeding with the advantage to be an office procedure without either risks and costs of D&C.

  4. Elastography as a new diagnostic tool to detect breast cancer – evaluation of research and clinical applications

    Directory of Open Access Journals (Sweden)

    Paweł Rzymski

    2011-10-01

    Full Text Available Mammography and ultrasonography are currently the most sensitive methods for detecting breast cancer,but elastography is a new diagnostic tool. Stiffness of invasive carcinomas were found to be 5-25 times largerthan that of normal adipose tissue. There are basically two types of elastography examination used in initialclinical research: conventional elastography with compression and shear wave elastography. Analysis of 20 studiesin the years 1997-2010 with 1484 malignant and 2822 benign breast lesions is presented in this paper. Theoverall sensitivity was 67-100% with specificity of 62-99% for elastography. This raises the hope of introducingthis method in the BI-RADS classification and modifying diagnostics in category 3-4.

  5. Trichomoniasis: a brief review of diagnostic methods and our experience with real-time PCR for detecting infection.

    Science.gov (United States)

    Šoba, Barbara; Skvarč, Miha; Matičič, Mojca

    2015-01-01

    Trichomoniasis is the most common non-viral sexually transmitted infection, and it is caused by the protozoan flagellate Trichomonas vaginalis. Although highly prevalent in sexually active women, it has long been overlooked in other groups of potentially infected people. Recently, studies have shown that trichomoniasis increases the risk of infection with human immunodeficiency virus and can cause adverse outcomes of pregnancy, which has increased interest in T. vaginalis and increased the need for highly sensitive diagnostic tests. This article summarizes the diagnostic methods most commonly used in the diagnosis of trichomoniasis, including the most sensitive and specific nucleic acid amplification tests. It also presents the results of our study comparing the performance of wet mount microscopy and culture to real-time PCR for detecting the parasite.

  6. Evaluation of yield of currently available diagnostics by sample type to optimize detection of respiratory pathogens in patients with a community-acquired pneumonia

    NARCIS (Netherlands)

    E. Huijskens (Elisabeth); J.W. Rossen (John); J.A.J.W. Kluytmans (Jan); A.G.M. van der Zanden (Adri); M.P.G. Koopmans D.V.M. (Marion)

    2014-01-01

    textabstractBackground: For the detection of respiratory pathogens, the sampling strategy may influence the diagnostic yield. Ideally, samples from the lower respiratory tract are collected, but they are difficult to obtain. Objectives: In this study, we compared the diagnostic yield in sputum and o

  7. Evaluation of yield of currently available diagnostics by sample type to optimize detection of respiratory pathogens in patients with a community-acquired pneumonia

    NARCIS (Netherlands)

    Huijskens, Elisabeth G. W.; Rossen, John W. A.; Kluytmans, Jan A. J. W.; van der Zanden, Adri G. M.; Koopmans, Marion

    2014-01-01

    BACKGROUND: For the detection of respiratory pathogens, the sampling strategy may influence the diagnostic yield. Ideally, samples from the lower respiratory tract are collected, but they are difficult to obtain. OBJECTIVES: In this study, we compared the diagnostic yield in sputum and oropharyngeal

  8. Evaluation of yield of currently available diagnostics by sample type to optimize detection of respiratory pathogens in patients with a community-acquired pneumonia

    NARCIS (Netherlands)

    Huijskens, Elisabeth G. W.; Rossen, John W. A.; Kluytmans, Jan A. J. W.; van der Zanden, Adri G. M.; Koopmans, Marion

    2014-01-01

    BACKGROUND: For the detection of respiratory pathogens, the sampling strategy may influence the diagnostic yield. Ideally, samples from the lower respiratory tract are collected, but they are difficult to obtain. OBJECTIVES: In this study, we compared the diagnostic yield in sputum and oropharyngeal

  9. Comparison of Digital Rectal and Microchip Transponder Thermometry in Ferrets (Mustela putorius furo)

    Science.gov (United States)

    Maxwell, Branden M; Brunell, Marla K; Olsen, Cara H; Bentzel, David E

    2016-01-01

    Body temperature is a common physiologic parameter measured in both clinical and research settings, with rectal thermometry being implied as the ‘gold standard.’ However, rectal thermometry usually requires physical or chemical restraint, potentially causing falsely elevated readings due to animal stress. A less stressful method may eliminate this confounding variable. The current study compared 2 types of digital rectal thermometers—a calibrated digital thermometer and a common digital thermometer—with an implantable subcutaneous transponder microchip. Microchips were implanted subcutaneously between the shoulder blades of 16 ferrets (8 male, 8 female), and temperatures were measured twice from the microchip reader and once from each of the rectal thermometers. Results demonstrated the microchip temperature readings had very good to good correlation and agreement to those from both of the rectal thermometers. This study indicates that implantable temperature-sensing microchips are a reliable alternative to rectal thermometry for monitoring body temperature in ferrets. PMID:27177569

  10. Diagnostic utility of zinc protoporphyrin to detect iron deficiency in Kenyan preschool children: a community-based survey.

    Science.gov (United States)

    Teshome, Emily M; Prentice, Andrew M; Demir, Ayşe Y; Andang'o, Pauline E A; Verhoef, Hans

    2017-01-01

    Zinc protoporphyrin (ZPP) has been used to screen and manage iron deficiency in individual children, but it has also been recommended to assess population iron status. The diagnostic utility of ZPP used in combination with haemoglobin concentration has not been evaluated in pre-school children. We aimed to a) identify factors associated with ZPP in children aged 12-36 months; b) assess the diagnostic performance and utility of ZPP, either alone or in combination with haemoglobin, to detect iron deficiency. We used baseline data from 338 Kenyan children enrolled in a community-based randomised trial. To identify factors related to ZZP measured in whole blood or erythrocytes, we used bivariate and multiple linear regression analysis. To assess diagnostic performance, we excluded children with elevated plasma concentrations of C-reactive protein or α1-acid glycoprotein, and with Plasmodium infection, and we analysed receiver operating characteristics (ROC) curves, with iron deficiency defined as plasma ferritin concentration iron deficiency. Whole blood ZPP and erythrocyte ZPP were independently associated with haemoglobin concentration, Plasmodium infection and plasma concentrations of soluble transferrin receptor, ferritin, and C-reactive protein. In children without inflammation or Plasmodium infection, the prevalence of true iron deficiency was 32.1%, compared to prevalence of 97.5% and 95.1% when assessed by whole blood ZPP and erythrocyte ZPP with conventional cut-off points (70 μmol/mol and 40 μmol/mol haem, respectively). Addition of whole blood ZPP or erythrocyte ZPP to haemoglobin concentration increased the area-under-the-ROC-curve (84.0%, p = 0.003, and 84.2%, p = 0.001, respectively, versus 62.7%). A diagnostic rule (0.038689 [haemoglobin concentration, g/L] + 0.00694 [whole blood ZPP, μmol/mol haem] >5.93120) correctly ruled out iron deficiency in 37.4%-53.7% of children screened, depending on the true prevalence, with both specificity and

  11. Combining in Vitro Diagnostics with in Vivo Imaging for Earlier Detection of Pancreatic Ductal Adenocarcinoma: Challenges and Solutions.

    Science.gov (United States)

    Laeseke, Paul F; Chen, Ru; Jeffrey, R Brooke; Brentnall, Teresa A; Willmann, Jürgen K

    2015-12-01

    Pancreatic ductal adenocarcinoma (PDAC) is the fourth-leading cause of cancer-related death in the United States and is associated with a dismal prognosis, particularly when diagnosed at an advanced stage. Overall survival is significantly improved if PDAC is detected at an early stage prior to the onset of symptoms. At present, there is no suitable screening strategy for the general population. Available diagnostic serum markers are not sensitive or specific enough, and clinically available imaging modalities are inadequate for visualizing early-stage lesions. In this article, the role of currently available blood biomarkers and imaging tests for the early detection of PDAC will be reviewed. Also, the emerging biomarkers and molecularly targeted imaging agents being developed to improve the specificity of current imaging modalities for PDAC will be discussed. A strategy incorporating blood biomarkers and molecularly targeted imaging agents could lead to improved screening and earlier detection of PDAC in the future. (©) RSNA, 2015.

  12. Detection of Chlamydia trachomatis in blood samples as a diagnostic method for complicated and persistent forms of urogenital chlamydia infections

    Directory of Open Access Journals (Sweden)

    Sultanakhmedov E.S.

    2015-09-01

    Full Text Available Goal: the study of the effectiveness of the method for laboratory diagnostics of urogenital chlamydial infection in patients with chronic form of the disease. Material and methods. The presence of DNAof C. trachomatis was detected by PCR in either genital or extragenital (blood sites in eighth patients (four men and four women. Results. It is established that in biological material taken from extragenital (blood sites, C. trachomatis was detected in all patients examined (in 100% of cases, while in clinical samples obtained from genital sites, in seven patients only (87.5%. Conclusion. We found that specific chlamydial DNAcan be detected in extragenital (blood site, despite the negative reaction in the clinical material from the genital tract of patients with genital chlamydial infection.

  13. Diagnostic value of DIAGNOdent in detecting caries under composite restorations of primary molars

    Directory of Open Access Journals (Sweden)

    Ava Vali Sichani

    2016-01-01

    Conclusion: DIAGNOdent showed a greater accuracy in detecting secondary caries under primary molar restorations, compared to radiographs. Although DIAGNOdent is an effective method for detecting caries under composite restorations, it is better to be used as an adjunctive method alongside other detecting procedures.

  14. Bacillus 'next generation' diagnostics: Moving from detection towards sub-typing and risk related strain profiling

    Directory of Open Access Journals (Sweden)

    Monika eEhling-Schulz

    2013-02-01

    Full Text Available The highly heterogeneous genus Bacillus comprises the largest species group of endospore forming bacteria. Because of their ubiquitous nature, Bacillus spores can enter food production at several stages resulting in significant economic losses and posing a potential risk to consumers due the capacity of certain Bacillus strains for toxin production. In the past, food microbiological diagnostics was focused on the determination of species using conventional culture based methods, which are still widely used. However, due to the extreme intraspecies diversity found in the genus Bacillus, DNA based identification and typing methods are gaining increasing importance in routine diagnostics. Several studies showed that certain characteristics are rather strain dependent than species specific. Therefore, the challenge for current and future Bacillus diagnostics is not only the efficient and accurate identification on species level but also the development of rapid methods to identify strains with specific characteristics (such as stress resistance or spoilage potential, trace contamination sources, and last but not least discriminate potential hazardous strains from non-toxic strains.

  15. Detection of mullerian duct anomalies: diagnostic utility of two dimensional ultrasonography as compared to magnetic resonance imaging

    Directory of Open Access Journals (Sweden)

    Krishna Pratap Singh Senger

    2016-12-01

    Full Text Available Background: Mullerian duct anomalies (MDAs are a fascinating group of disorders that have varied clinical presentation from being asymptomatic to primary amenorrhea to inability to reproduce. Correct diagnosis of the condition plays a crucial role in management. Imaging plays a pivotal role in making correct diagnosis. This study aims to find the prevalence of MDAs amongst study population and their relation with infertility and also compares diagnostic utility of pelvic ultrasound with MRI. Methods: A randomized diagnostic test evaluation study was conducted in the Department of Radiodiagnosis and Imaging of a tertiary care teaching hospital over a period of 2 years. The patient first underwent pelvic 2D USG in multiple planes using curvilinear probe of 3MHz to 5 MHz. frequency and then MRI. Results: Most common MDA in total study sample and in primary infertility group is arcuate uterus while in recurrent abortions group it is unicornuate uterus. Out of total study sample of 75 patients 2D USG detected 18 cases of MDA while MRI detected 22 cases of MDA. So, 2D USG failed to detect 04 cases of MDA in total study population bringing overall sensitivity of 2D USG as 81.8%, specificity of 100%, PPV of 100%, NPV of 93.4% and accuracy of 94.6%. Conclusions: 2D USG has a few limitations but in view of relatively simple imaging procedure, ease of availability and cost effectiveness it should be utilized as an initial imaging modality in patients with suspicion of MDAs.

  16. Discrimination of Bacillus anthracis from closely related microorganisms by analysis of 16S and 23S rRNA with oligonucleotide microchips

    Science.gov (United States)

    Bavykin, Sergei G.; Mirzabekov, Andrei D.

    2007-10-30

    The present invention is directed to a novel method of discriminating a highly infectious bacterium Bacillus anthracis from a group of closely related microorganisms. Sequence variations in the 16S and 23S rRNA of the B. cereus subgroup including B. anthracis are utilized to construct an array that can detect these sequence variations through selective hybridizations. The identification and analysis of these sequence variations enables positive discrimination of isolates of the B. cereus group that includes B. anthracis. Discrimination of single base differences in rRNA was achieved with a microchip during analysis of B. cereus group isolates from both single and in mixed probes, as well as identification of polymorphic sites. Successful use of a microchip to determine the appropriate subgroup classification using eight reference microorganisms from the B. cereus group as a study set, was demonstrated.

  17. Stacking-cyclodextrin-microchip electrokinetic chromatographic determination of gabapentinoid drugs in pharmaceutical and biological matrices.

    Science.gov (United States)

    Zeid, Abdallah M; Kaji, Noritada; Nasr, Jenny Jeehan M; Belal, Fathalla F; Baba, Yoshinobu; Walash, Mohamed I

    2017-06-23

    A facile, rapid, and highly sensitive microchip-based electrokinetic chromatographic method was developed for the simultaneous analysis of two gabapentinoid drugs, gabapentin (GPN) and pregabalin (PGN). Both drugs were first reacted with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) via nucleophilic substitution reactions to yield highly fluorescent products with λex/em 470/540nm. Analyses of both fluorescently labeled compounds were achieved within 200s in a poly(methyl methacrylate) (PMMA) microchip with a 30mm separation channel. Optimum separation was achieved using a borate buffer (pH 9.0) solution containing methylcellulose and β-cyclodextrin (β-CD) as buffer additives. Methylcellulose acted as a dynamic coating to prevent adsorption of the studied compounds on the inner surfaces of the microchannels, while β-CD acted as a pseudo-stationary phase to improve the separation efficiency between the labeled drugs with high resolution (Rs>7). The fluorescence intensities of the labeled drugs were measured using a light emitting diode-induced fluorescence detector at 540nm after excitation at 470nm. The sensitivity of the method was enhanced 14- and 17-fold for PGN and GPN, respectively by field-amplified stacking relative to traditional pinched injection so that it could quantify 10ngmL(-1) for both analytes, with a detection limit lower than 3ngmL(-1). The developed method was efficiently applied to analyze PGN and GPN in their pharmaceutical dosage forms and in biological fluids. The extraction recoveries of the studied drugs from plasma and urine samples were more than 89% with%RSD values lower than 6.2. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Continuous Monitoring of Nitrate and Sulfate in Aerosols with Microchip Electrophoresis

    Science.gov (United States)

    Noblitt, S. D.; Henry, C. S.; Collett, J. L.; Hering, S. V.

    2007-12-01

    Routine monitoring of aerosol composition is important since aerosols can negatively affect both the environment and health. Water-soluble inorganic ions are commonly monitored using the particle-into-liquid-sampler coupled to ion chromatography (PILS-IC). However, a less-expensive, faster, and more portable analysis system is desirable. Here, we present the coupling of microchip capillary electrophoresis (MCE) to a water-based condensation particle counter (WCPC) for rapid and continuous monitoring of chloride, nitrate, and sulfate in atmospheric aerosols. To achieve a working system, several obstacles were overcome. A working interface between the electrophoresis microchip and the WCPC sampler was developed. This interface was designed to remove insoluble particles from the analysis stream and to prevent the sampling-induced pressure gradient from altering flow in the microfluidic device. The electrophoresis separation chemistry was optimized for the small chip size, to be free from potential interfering compounds, and to operate continuously for several hours. In-field performance of the integrated system was tested with ambient aerosols. Anion analyses can be performed in less than two minutes with aerosol detection limits similar to the PILS-IC, but with greater portability and reduced cost. Coupling microfluidic devices to aerosol sampling technology proves successful for inorganic anion analysis and shows potential for faster and more sensitive measurements as well as monitoring of other water- soluble aerosol components such as organic acids, cations, and carbohydrates. The reduced cost and size relative to current technology indicate that greater deployment of monitoring stations or the advent of portable analyzers may be feasible.

  19. Diagnostic accuracy of in-stent coronary restenosis detection with multislice spiral computed tomography: a meta-analysis

    Energy Technology Data Exchange (ETDEWEB)

    Hamon, Michele [University Hospital of Caen, Department of Radiology, 14033 Caen Cedex, Normandy (France); Champ-Rigot, Laure; Riddell, John W. [University Hospital of Caen, Department of Cardiology, 14033 Caen Cedex, Normandy (France); Morello, Remy [University Hospital of Caen, Department of Statistics, 14033 Caen Cedex, Normandy (France); Hamon, Martial [University Hospital of Caen, Department of Cardiology, 14033 Caen Cedex, Normandy (France); Institut Pasteur de Lille, INSERM 744, Lille (France); Centre Hospitalier Universitaire de Caen, Service des Maladies du Coeur et des Vaisseaux, 14033 Caen Cedex, Normandy (France)

    2008-02-15

    This study was designed to define the current role of multislice spiral computed tomography (MSCT) for the diagnosis of coronary in-stent restenosis using a meta-analytic process. Restenosis remains a limitation after coronary stent implantation and contributes to a substantial number of coronary re-assessments by conventional invasive coronary angiography (CA). We identified 15 studies (807 patients) evaluating in-stent restenosis by means of both MSCT ({>=}16 slices) and conventional CA until February 2007. After data extraction the analysis was performed according to a random-effects model. The analysis pooled the results from 15 studies with a total of 1,175 stents. A substantial number of unassessable stents (13%) were excluded from the analysis underscoring the shortcomings of MSCT. With this major limitation the diagnostic performance of MSCT for in-stent restenosis detection can be summarized as follows: the sensitivity and specificity were 84% [95% confidence interval (CI) 77-89%] and 91% (95% CI 89-93%), respectively, with positive and negative likelihood ratios of 12.2 (95% CI 6.6-22.6) and 0.23 (95% CI 0.17-0.31), respectively, and with a diagnostic odds ratio of 67.9 (95% CI 34.4-134.1). MSCT has shortcomings difficult to overcome in daily practice for in-stent restenosis detection and continues to have moderately high sensitivity and specificity. The diagnostic role of this emerging technology as an alternative to CA for in-stent restenosis detection remains limited. (orig.)

  20. Development and evaluation of a saliva-based chair-side diagnostic for the detection of Porphyromonas gingivalis

    Directory of Open Access Journals (Sweden)

    Neil M. O'Brien-Simpson

    2015-09-01

    Full Text Available Porphyromonas gingivalis is a key pathogen in the polymicrobial biofilm that is associated with the oral disease chronic periodontitis. A number of studies have shown that in humans the level of P. gingivalis in the polymicrobial biofilm is positively correlated with disease progression. The aim of this study was to develop a P. gingivalis diagnostic that has high specificity and sensitivity for P. gingivalis using a range of laboratory and clinical isolates and then compare the efficacy of the diagnostic with RTPCR using samples from chronic periodontitis patients and age- and sex-matched healthy controls. Key parameters for the kit were to use saliva as the biological fluid as this is a most convenient medium for chair-side sampling and to give a positive reading for the reported threshold for detection of 5×105 P. gingivalis cells/mL that indicates disease progression. We initially screened a range of monoclonal antibodies for recognition of the P. gingivalis conserved virulence factor RgpA-Kgp complex and identified two mAbs that could be used in a capture and detection ELISA system. These mAbs were used to formulate and manufacture the GC P. gingivalis saliva diagnostic kit used in the study. To validate the saliva kit, saliva (P. gingivalis free was spiked with known concentrations of viable P. gingivalis whole cells of W50, 381, A7A1-28, and ATCC 33277; P. gingivalis clinical isolates; P. gingivalis vesicles; and the secreted form of the RgpA-Kgp complex. Laboratory findings indicated that the kit was able to detect all laboratory and clinical isolate strains of P. gingivalis at 5×104/mL to 5×105/mL. It was also able to detect the RgpA-Kgp complex and vesicles at 5×104 and 5×105 cell equivalent doses, respectively. Saliva and plaque were then collected from 50 subjects with moderate–severe chronic periodontitis and 50 age- and sex-matched subjects with healthy periodontium. Real-time PCR was utilised to analyse levels of P

  1. Development and evaluation of a saliva-based chair-side diagnostic for the detection of Porphyromonas gingivalis.

    Science.gov (United States)

    O'Brien-Simpson, Neil M; Burgess, Kate; Brammar, Gail C; Darby, Ivan B; Reynolds, Eric C

    2015-01-01

    Porphyromonas gingivalis is a key pathogen in the polymicrobial biofilm that is associated with the oral disease chronic periodontitis. A number of studies have shown that in humans the level of P. gingivalis in the polymicrobial biofilm is positively correlated with disease progression. The aim of this study was to develop a P. gingivalis diagnostic that has high specificity and sensitivity for P. gingivalis using a range of laboratory and clinical isolates and then compare the efficacy of the diagnostic with RTPCR using samples from chronic periodontitis patients and age- and sex-matched healthy controls. Key parameters for the kit were to use saliva as the biological fluid as this is a most convenient medium for chair-side sampling and to give a positive reading for the reported threshold for detection of 5×10(5) P. gingivalis cells/mL that indicates disease progression. We initially screened a range of monoclonal antibodies for recognition of the P. gingivalis conserved virulence factor RgpA-Kgp complex and identified two mAbs that could be used in a capture and detection ELISA system. These mAbs were used to formulate and manufacture the GC P. gingivalis saliva diagnostic kit used in the study. To validate the saliva kit, saliva (P. gingivalis free) was spiked with known concentrations of viable P. gingivalis whole cells of W50, 381, A7A1-28, and ATCC 33277; P. gingivalis clinical isolates; P. gingivalis vesicles; and the secreted form of the RgpA-Kgp complex. Laboratory findings indicated that the kit was able to detect all laboratory and clinical isolate strains of P. gingivalis at 5×10(4)/mL to 5×10(5)/mL. It was also able to detect the RgpA-Kgp complex and vesicles at 5×10(4) and 5×10(5) cell equivalent doses, respectively. Saliva and plaque were then collected from 50 subjects with moderate-severe chronic periodontitis and 50 age- and sex-matched subjects with healthy periodontium. Real-time PCR was utilised to analyse levels of P. gingivalis in both

  2. Detection of All Species of the Genus Alphavirus by Reverse Transcription-PCR with Diagnostic Sensitivity▿

    OpenAIRE

    Grywna, K.; Kupfer, B.; Panning, M.; Drexler, J. F.; Emmerich, P.; Drosten, C.; Kummerer, B. M.

    2010-01-01

    Clinical arbovirus screening requires exclusion of a broad range of viruses with as few assays as possible. We present a reverse transcription-PCR (RT-PCR) for the detection of all species of the genus Alphavirus qualified for exclusion screening (limit of detection [LOD], 5 to 100 RNA copies per reaction across all Alphavirus species; detection of viremia down to ca. 10,000 copies per ml).

  3. Diagnostic Accuracy of Digital and Conventional Radiography in the Detection of Non-Cavitated Approximal Dental Caries

    Directory of Open Access Journals (Sweden)

    Farida Abesi

    2012-03-01

    Full Text Available Background: Radiography plays an important role in the detection of interproximal caries.Objectives: The aim of the present study was to determine diagnostic accuracy of chargecoupled devices (CCD, Photo Stimulable Phosphor (PSP and film radiography in detecting non-cavitated caries.Patients and Methods: Seventy-two non-cavitated approximal surfaces of extracted human posterior teeth were radiographed under standardized conditions using three intraoral modalities: CCD Dixi3 (Planmeca, Finland, PSP Digora PCT (Soredex, Finland, and E-speed film (Kodak, USA. Radiographs were interpreted by four observers and caries lesions were classified as sound (R0, restricted to enamel (R1, reaching the dentinoenamel junction (DEJ and the outer half of the dentin (R2 and the inner half of the dentin (R3. The teeth were subsequently sectioned for histological analysis which served as the gold standard for radiographic examination.Results: Microscopic examinations showed that the distribution of caries were 63.9% sound, 18.1% enamel, 9.7% DEJ and outer half of the dentin and 8.3% into the inner half of the dentin.The sensitivity and specificity of film, CCD and PSP for the detection of enamel caries were 38% and 98%; 15% and 96 %; and 23% and 98%, respectively. The sensitivity and specificity of film, CCD and PSP for the detection of both dentin and enamel caries were 55% and 100%; 45% and 100% ; and 55% and 100%, respectively.Conclusions: The results demonstrated that the diagnostic accuracy of digital images is similar to that of conventional film radiography in the detection of non-cavitated approximal caries.

  4. Real-Time Reverse Transcription PCR Assay for Detection of Senecavirus A in Swine Vesicular Diagnostic Specimens.

    Directory of Open Access Journals (Sweden)

    Alexa J Bracht

    Full Text Available Senecavirus A (SV-A, formerly, Seneca Valley virus (SVV, has been detected in swine with vesicular lesions and is thought to be associated with swine idiopathic vesicular disease (SIVD, a vesicular disease syndrome that lacks a defined causative agent. The clinical presentation of SIVD resembles that of other more contagious and economically devastating vesicular diseases, such as foot-and-mouth disease (FMD, swine vesicular disease (SVD, and vesicular stomatitis (VS, that typically require immediate rule out diagnostics to lift restrictions on animal quarantine, movement, and trade. This study presents the development of a sensitive, SYBR Green RT-qPCR assay suitable for detection of SV-A in diagnostic swine specimens. After testing 50 pigs with clinical signs consistent with vesicular disease, 44 (88% were found to be positive for SV-A by RT-qPCR as compared to none from a negative cohort of 35 animals without vesicular disease, indicating that the assay is able to successfully detect the virus in an endemic population. SV-A RNA was also detectable at a low level in sera from a subset of pigs that presented with (18% or without (6% vesicular signs. In 2015, there has been an increase in the occurrence of SV-A in the US, and over 200 specimens submitted to our laboratory for vesicular investigation have tested positive for the virus using this method. SV-A RNA was detectable in all common types of vesicular specimens including swabs and tissue from hoof lesions, oral and snout epithelium, oral swabs, scabs, and internal organ tissues such as liver and lymph node. Genome sequencing analysis from recent virus isolates was performed to confirm target amplicon specificity and was aligned to previous isolates.

  5. Evaluation of annealing and double ion beam irradiation by a laser-induced and laser-detected surface acoustic wave diagnostic system

    Science.gov (United States)

    Kitazawa, Sin-iti; Wakai, Eiichi; Aoto, Kazumi

    2016-10-01

    The effects of annealing and double ion irradiation on nuclear structural materials were investigated using a novel, non-destructive, non-contact diagnostic method. A laser-induced and laser-detected surface acoustic wave (SAW) was adopted as a diagnostic system. The SAWs propagation velocity and the SAWs vibration velocity along the normal direction of the surface were measured to investigate mechanical properties of the substrates. Change of the shear modulus was detected in the annealed substrates. Non-linear effect on amplitude of the excited SAW was observed on the double ion irradiated materials. The potential of the SAW diagnostic system for assessing nuclear structural materials was demonstrated.

  6. General and advanced diagnostic tools to detect Mycobacterium tuberculosis and their drug susceptibility: a review

    NARCIS (Netherlands)

    Gazi, M.A.; Islam, M.R.; Kibria, M.; Mahmud, Z.

    2015-01-01

    The global control of tuberculosis remains a great challenge from the standpoint of diagnosis, detection of drug resistance, and treatment, because treatment can only be initiated when infection is detected, and is guided by the results of antimicrobial susceptibility testing. To a large extent, non

  7. Evaluation of the currently used diagnostic procedures for the detection of Brucella melitensis in sheep

    NARCIS (Netherlands)

    Bercovich, Z.; Guler, L.; Baysal, T.; Schreuder, B.E.C.; Zijderveld, van F.G.

    1998-01-01

    A study was conducted to determine whether the use of the enzyme-linked immunosorbent assay (ELISA) improves detection of brucellosis in individual sheep. Sera from 132 sheep that aborted due to B. melitensis were used to assess the efficacy of the ELISA to detect brucellosis in sheep. ELISA results

  8. Hepcidin detects iron deficiency in Sri Lankan adolescents with a high burden of hemoglobinopathy: A diagnostic test accuracy study

    Science.gov (United States)

    Wray, Katherine; Allen, Angela; Evans, Emma; Fisher, Chris; Premawardhena, Anuja; Perera, Lakshman; Rodrigo, Rexan; Goonathilaka, Gayan; Ramees, Lebbe; Webster, Craig; Armitage, Andrew E; Prentice, Andrew M

    2017-01-01

    Abstract Anemia affects over 800 million women and children globally. Measurement of hepcidin as an index of iron status shows promise, but its diagnostic performance where hemoglobinopathies are prevalent is unclear. We evaluated the performance of hepcidin as a diagnostic test of iron deficiency in adolescents across Sri Lanka. We selected 2273 samples from a nationally representative cross‐sectional study of 7526 secondary schoolchildren across Sri Lanka and analyzed associations between hepcidin and participant characteristics, iron indices, inflammatory markers, and hemoglobinopathy states. We evaluated the diagnostic accuracy of hepcidin as a test for iron deficiency with estimation of the AUCROC, sensitivity/specificity at each hepcidin cutoff, and calculation of the Youden Index to find the optimal threshold. Hepcidin was associated with ferritin, sTfR, and hemoglobin. The AUCROC for hepcidin as a test of iron deficiency was 0.78; hepcidin outperformed Hb and sTfR. The Youden index‐predicted cutoff to detect iron deficiency (3.2 ng/mL) was similar to thresholds previously identified to predict iron utilization and identify deficiency in African populations. Neither age, sex, nor α‐ or β‐thalassemia trait affected diagnostic properties of hepcidin. Hepcidin pre‐screening would prevent most iron‐replete thalassemia carriers from receiving iron whilst still ensuring most iron deficient children were supplemented. Our data indicate that the physiological relationship between hepcidin and iron status transcends specific populations. Measurement of hepcidin in individuals or populations could establish the need for iron interventions. PMID:27883199

  9. Is diagnostic accuracy for detecting pulmonary nodules in chest CT reduced after a long day of reading?

    Science.gov (United States)

    Krupinski, Elizabeth A.; Berbaum, Kevin S.; Caldwell, Robert; Schartz, Kevin M.

    2012-02-01

    Radiologists are reading more cases with more images, especially in CT and MRI and thus working longer hours than ever before. There have been concerns raised regarding fatigue and whether it impacts diagnostic accuracy. This study measured the impact of reader visual fatigue by assessing symptoms, visual strain via dark focus of accommodation, and diagnostic accuracy. Twenty radiologists and 20 radiology residents were given two diagnostic performance tests searching CT chest sequences for a solitary pulmonary nodule before (rested) and after (tired) a day of clinical reading. 10 cases used free search and navigation, and the other 100 cases used preset scrolling speed and duration. Subjects filled out the Swedish Occupational Fatigue Inventory (SOFI) and the oculomotor strain subscale of the Simulator Sickness Questionnaire (SSQ) before each session. Accuracy was measured using ROC techniques. Using Swensson's technique yields an ROC area = 0.86 rested vs. 0.83 tired, p (one-tailed) = 0.09. Using Swensson's LROC technique yields an area = 0.73 rested vs. 0.66 tired, p (one-tailed) = 0.09. Using Swensson's Loc Accuracy technique yields an area = 0.77 rested vs. 0.72 tired, p (one-tailed) = 0.13). Subjective measures of fatigue increased significantly from early to late reading. To date, the results support our findings with static images and detection of bone fractures. Radiologists at the end of a long work day experience greater levels of measurable visual fatigue or strain, contributing to a decrease in diagnostic accuracy. The decrease in accuracy was not as great however as with static images.

  10. STUDY OF CAPILLARY ELECTROPHORESIS ON MICROCHIP BASED ON MEMS

    Institute of Scientific and Technical Information of China (English)

    Wang Ming; Li Wei; Han Jinghong; Cui Dafu

    2002-01-01

    Using a standard photolithographical procedure, chemical wet etching and thermal diffusion bonding technology, a chemical analysis device for Capillary Electrophoresis(CE) has been microfabricated on a planar glass substrate with a cross-column geometry. The channels on the microchip substrate are about 50μm deep and 150μm wide. By employing amino acids derived from 2,4-DiNitroFluoroBenzen (DNFB) on CE chip channels, the sample manipulating system is studied based on the principle of electrodynamics.

  11. STUDY OF CAPILLARY ELECTROPHORESIS ON MICROCHIP BASED ON MEMS

    Institute of Scientific and Technical Information of China (English)

    WangMing; LiWei; 等

    2002-01-01

    Using a standard photolithographical procedure,chenmical wet etching and thermal diffusion bonding technology,a chemical analysis device for Capillary Electrophoresis(CE) has been microfabricated on a planar glass substrate with a cross-column geometry.The channels on the microchip substrate are about 50um deep and 150um wide.By employing amino acids derived from 2,4-DiNitroFluoroBenzen(DNFB) on CE chip channels,the sample manipulating system is studied based on the principle of electrodynamics.

  12. A disposable laser print-cut-laminate polyester microchip for multiplexed PCR via infra-red-mediated thermal control

    Energy Technology Data Exchange (ETDEWEB)

    Ouyang, Yiwen [Department of Chemistry, University of Virginia, Charlottesville, VA 22904 (United States); Duarte, Gabriela R.M. [Department of Chemistry, University of Virginia, Charlottesville, VA 22904 (United States); Universidade Federal de Goiás, Goiânia, GO 74690-900 (Brazil); Poe, Brian L.; Riehl, Paul S. [Department of Chemistry, University of Virginia, Charlottesville, VA 22904 (United States); Santos, Fernando M. dos; Martin-Didonet, Claudia C.G. [Universidade Estadual de Goiás, Anápolis, GO 75132-400 (Brazil); Carrilho, Emanuel [Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos, SP 13566-590 (Brazil); Instituto Nacional de Ciência e Tecnologia de Bioanalítica, CP 6154, Campinas, SP 13083-970 (Brazil); Landers, James P., E-mail: landers@virginia.edu [Department of Chemistry, University of Virginia, Charlottesville, VA 22904 (United States); Department of Mechanical Engineering, University of Virginia, Charlottesville, VA 22904 (United States); Department of Pathology, University of Virginia Health Science Center, Charlottesville, VA (United States)

    2015-12-11

    Infrared (IR)-mediated thermal cycling system, a method proven to be a effective for sub-μL scale polymerase chain reaction (PCR) on microchips, has been integrated with DNA extraction and separation on a glass microchip in a fully integrated micro Total Analysis System by Easley et al., in 2006. IR-PCR has been demonstrated on both glass and PMMA microdevices where the fabrication (bonding) is not trivial. Polyester-toner (PeT) microfluidic devices have significant potential as cost-effective, disposable microdevices as a result of the ease of fabrication (∼$0.25 USD and <10 min per device) and availability of commercial substrates. For the first time, we demonstrate here the thermal cycling in PeT microchips on the IR-PCR system. Undesirable IR absorption by the black-toner bonding layer was eliminated with a spatial filter in the form of an aluminum foil mask. The solution heating rate for a black PeT microchip using a tungsten lamp was 10.1 ± 0.7 °C s{sup −1} with a cooling rate of roughly −12 ± 0.9 °C s{sup −1} assisted by forced air cooling. Dynamic surface passivation strategies allowed the successful amplification of a 520 bp fragment of the λ-phage genome (in 11 min) and a 1500 bp region of Azospirillum brasilense. Using a centrosymmetric chamber configuration in a multichamber PeT microchip, homogenous temperature distribution over all chambers was achieved with inter-chamber temperature differences at annealing, extension and denaturing steps of less than ±2 °C. The effectiveness of the multichamber system was demonstrated with the simultaneous amplification of a 390 bp amplicon of human β-globin gene in five PeT PCR microchambers. The relative PCR amplification efficiency with a human β-globin DNA fragment ranged from 70% to 90%, in comparison to conventional thermal cyclers, with an inter-chamber standard deviation of ∼10%. Development of PeT microchips for IR-PCR has the potential to provide rapid, low

  13. Impact of uncontrolled blood pressure on diagnostic accuracy of coronary flow reserve for detecting significant coronary stenosis in hypertensive patients

    Institute of Scientific and Technical Information of China (English)

    LI Wei-hong; XU Wei-xian; LI Zhao-ping; LI Cui-ping; WANG Xin-yu; HE Li-yun; ZHAO Wei

    2013-01-01

    Baciground Impaired coronary flow reserve (CFR) in patients with hypertension may be caused by epicardial coronary stenosis or microvascular dysfunction.Antihypertensive treatment has been shown to improve coronary microvascular dysfunction.The aim of this study was to evaluate the impact of uncontrolled blood pressure (BP) on diagnostic accuracy of CFR for detecting significant coronary stenosis.Methods A total of 98 hypertensive patients scheduled for coronary angiography (CAG) due to chest pain were studied.Of them,45 patients had uncontrolled BP (defined as the office BP >140/90 mmHg (1 mmHg=0.133 kPa) in general hypertensive patients,or >130/80 mmHg in hypertensive individuals with diabetes mellitus),and the remaining 53 patients had well-controlled BP.CFR was measured in the left anterior descending coronary artery (LAD) during adenosine triphosphate-induced hyperemia by non-invasive transthoracic Doppler echocardiography (TTDE) within 48 hours prior to CAG.Significant LAD stenosis was defined as >70% luminal narrowing.Diagnostic accuracy of CFR for detecting significant coronary stenosis was analyzed with a receiver operating characteristic analysis.Results CFR was significantly lower in patients with uncontrolled BP than in those with well-controlled BP (2.1±0.6 vs.2.6±0.9,P <0.01).Multivariate linear regression analysis of the study showed that the value of CFR was independently associated with the angiographically determined degree of LAD stenosis (β=-0.445,P <0.0001) and the presence of uncontrolled BP (β=-0.272,P=0.014).With a receiver operating characteristic analysis,CFR <2.2 was the optimal cut-off value for detecting LAD stenosis in all hypertensive patients (AUC 0.83,95%C/0.75-0.91) with a sensitivity of 75%,a specificity of 78%,and an accuracy of 77%.A significant reduction of diagnostic specificity was observed in patients with uncontrolled BP compared with those with well-controlled BP (67% vs.93%,P=0

  14. Development of rapid, sensitive and non-radioactive tissue-blot diagnostic method for the detection of citrus greening.

    Science.gov (United States)

    Nageswara-Rao, Madhugiri; Miyata, Shin-Ichi; Ghosh, Dilip; Irey, Mike; Garnsey, Stephen M; Gowda, Siddarame

    2013-01-01

    Citrus huanglongbing (HLB or citrus greening) is one of the most devastating diseases of citrus worldwide. The disease is caused by Gram-negative, phloem-limited α-proteobacterium, 'Candidatus Liberibacter asiaticus', vectored by the psyllid, Diaphorina citri Kuwayama. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection and non-uniform distribution within the tree makes the detection of the pathogen very difficult. Efficient management of HLB disease requires rapid and sensitive detection early in the infection followed by eradication of the source of pathogen and the vector. The polymerase chain reaction (PCR) based method is most commonly employed for screening the infected/suspected HLB plants and psyllids. This is time consuming, cumbersome and not practical for screening large number of samples in the field. To overcome this, we developed a simple, sensitive, non-radioactive, tissue-blot diagnostic method for early detection and screening of HLB disease. Digoxigenin labeled molecular probes specific to 'Ca. L. asiaticus' nucleotide sequences have been developed and used for the detection of the pathogen of the HLB disease. The copy number of the target genes was also assessed using real-time PCR experiments and the optimized real-time PCR protocol allowed positive 'Ca. L. asiaticus' detection in citrus samples infected with 'Ca. L. asiaticus' bacterium.

  15. Comparison of four diagnostic techniques for detection of Trichomonas vaginalis infection in females attending tertiary care hospital of North India

    Directory of Open Access Journals (Sweden)

    Razia Khatoon

    2015-01-01

    Full Text Available Background: Trichomonas vaginalis causes a common sexually transmitted disease trichomoniasis, which may lead to increased risk of transmission of human immunodeficiency virus infection and other pelvic inflammatory diseases. Wet mount examination is the most common test for diagnosis, but it has low sensitivity. Acridine orange staining can be used for diagnosis, but it requires special microscopic facility. Culture is considered as the gold standard, but it takes a long time for diagnosis. OSOM Trichomonas Rapid Test is a recently introduced rapid method based on immunochromatographic assay of trichomonal protein antigens. Hence, the present study was done to compare these four diagnostic techniques for detection of trichomoniasis in females with vaginal discharge. Materials and Methods: Vaginal swabs were taken from 835 female patients and wet mount examination, acridine orange staining, culture in Kupferberg medium, and OSOM Trichomonas Rapid Test, were performed. Results: Out of 835 patients included in our study, 68 (8.1% positive cases of trichomoniasis were detected by culture. OSOM Trichomonas Rapid Test detected 63 (7.5% cases, acridine orange staining detected 53 (6.3% cases, whereas, wet mount examination detected only 45 (5.4% positive cases. OSOM Trichomonas Rapid Test performed well and showed high sensitivity and specificity of 88.2% and 99.6%, respectively. Conclusion: As OSOM Trichomonas Rapid Test is a point of care test and gave better results than both wet mount examination and acridine orange staining; it can be used as a routine test in peripheral areas lacking laboratory facilities.

  16. Detection of ESAT-6 by a label free miniature immuno-electrochemical biosensor as a diagnostic tool for tuberculosis.

    Science.gov (United States)

    Diouani, Mohamed Fethi; Ouerghi, Oussama; Refai, Amira; Belgacem, Kamel; Tlili, Chaker; Laouini, Dhafer; Essafi, Makram

    2017-05-01

    Tuberculosis is a worldwide disease considered as a major health problem with high morbidity and mortality rates. Poor detection of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis remains a major obstacle to the global control of this disease. Here we report the development of a new test based on the detection of the major virulent factor of Mtb, namely the early secreted antigenic target 6-kDa protein or ESAT-6. A label free electrochemical immunosensor using an anti-ESAT-6 monoclonal antibody as a bio-receptor is described herein. Anti-ESAT-6 antibodies were first covalently immobilized on the surface of a gold screen-printed electrode functionalized via a self-assembled thiol monolayer. Interaction between the bio-receptor and ESAT-6 antigen was evaluated by square wave voltammetry method using [Fe(CN)6](3-/4-) as redox probe. The detection limit of ESAT-6 antigen was 7ng/ml. The immunosensor has also been able to detect native ESAT-6 antigen secreted in cell culture filtrates of three pathogenic strains of Mtb (CDC1551, H37RV and H8N8). Overall, this work describes an immune-electrochemical biosensor, based on ESAT-6 antigen detection, as a useful diagnostic tool for tuberculosis. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Evaluation of a commercial Erns-capture ELISA for detection of BVDV in routine diagnostic cattle serum samples

    Directory of Open Access Journals (Sweden)

    Ståhl Karl

    2007-03-01

    Full Text Available Abstract Background Bovine viral diarrhoea virus (BVDV is an important pathogen in cattle. The ability of the virus to cross the placenta during early pregnancy can result in the birth of persistently infected (PI calves. These calves shed the virus during their entire lifespan and are the key transmitters of infection. Consequently, identification (and subsequent removal of PI animals is necessary to rapidly clear infected herds from the virus. The objective of this study was to evaluate the suitability of a commercial Erns-capture ELISA, in comparison to the indirect immunoperoxidase test (IPX, for routine diagnostic detection of BVDV within a control programme. In addition, the effect of passive immunity and heat-inactivation of the samples on the performance of the ELISA was studied. Methods In the process of virus clearance within the Swedish BVDV control programme, all calves born in infected herds are tested for virus and antibodies. From such samples, sent in for routine diagnostics to SVA, we selected 220 sera collected from 32 beef herds and 29 dairy herds. All sera were tested for BVDV antigen using the Erns ELISA, and the results were compared to the results from the IPX used within the routine diagnostics. Results All 130 samples categorized as virus negative by IPX were tested negative in the ELISA, and all 90 samples categorized as virus positive were tested positive, i.e. the relative sensitivity and specificity of the ELISA was 100% in relation to IPX, and the agreement between the tests was perfect. Conclusion We can conclude that the Erns ELISA is a valid alternative that has several advantages compared to IPX. Our results clearly demonstrate that it performs well under Swedish conditions, and that its performance is comparable with the IPX test. It is highly sensitive and specific, can be used for testing of heat-inactivated samples, precolostral testing, and probably to detect PI animals at an earlier age than the IPX.

  18. Factors affecting the sensitivity and detection limits of MRI, CT, and SPECT for multimodal diagnostic and therapeutic agents.

    Science.gov (United States)

    Seevinck, Peter R; Seppenwoolde, Jan-Henry; de Wit, Tim C; Nijsen, Johannes F W; Beekman, Freek J; van Het Schip, Alfred D; Bakker, Chris J G

    2007-05-01

    Noninvasive imaging techniques like magnetic resonance imaging (MRI), computed tomography (CT) and single photon emission computed tomography (SPECT) play an increasingly important role in the diagnostic workup and treatment of cancerous disease. In this context, a distinct trend can be observed towards the development of contrast agents and radiopharmaceuticals that open up perspectives on a multimodality imaging approach, involving all three aforementioned techniques. To promote insight into the potentialities of such an approach, we prepared an overview of the strengths and limitations of the various imaging techniques, in particular with regard to their capability to quantify the spatial distribution of a multimodal diagnostic agent. To accomplish this task, we used a two-step approach. In the first step, we examined the situation for a particular therapeutic anti-cancer agent with multimodal imaging opportunities, viz. holmium-loaded microspheres (HoMS). Physical phantom experiments were performed to enable a comparative evaluation of the three modalities assuming the use of standard equipment, standard clinical scan protocols, and signal-known-exactly conditions. These phantom data were then analyzed so as to obtain first order estimates of the sensitivity and detection limits of MRI, CT and SPECT for HoMS. In the second step, the results for HoMS were taken as a starting point for a discussion of the factors affecting the sensitivity and detection limits of MRI, CT and SPECT for multimodal agents in general. In this, emphasis was put on the factors that must be taken into account when extrapolating the findings for HoMS to other diagnostic tasks, other contrast agents, other experimental conditions, and other scan protocols.

  19. Detection of subtelomere imbalance using MLPA: validation, development of an analysis protocol, and application in a diagnostic centre

    Directory of Open Access Journals (Sweden)

    Hills Alison

    2007-03-01

    Full Text Available Abstract Background Commercial MLPA kits (MRC-Holland are available for detecting imbalance at the subtelomere regions of chromosomes; each kit consists of one probe for each subtelomere. Methods For validation of the kits, 208 patients were tested, of which 128 were known to be abnormal, corresponding to 8528 genomic regions overall. Validation samples included those with trisomy 13, 18 and 21, microscopically visible terminal deletions and duplications, sex chromosome abnormalities and submicroscopic abnormalities identified by multiprobe FISH. A robust and sensitive analysis system was developed to allow accurate interpretation of single probe results, which is essential as breakpoints may occur between MLPA probes. Results The validation results showed that MLPA is a highly efficient technique for medium-throughput screening for subtelomere imbalance, with 95% confidence intervals for positive and negative predictive accuracies of 0.951-0.996 and 0.9996-1 respectively. A diagnostic testing strategy was established for subtelomere MLPA and any subsequent follow-up tests that may be required. The efficacy of this approach was demonstrated during 15 months of diagnostic testing when 455 patients were tested and 27 (5.9% abnormal cases were detected. Conclusion The development of a robust, medium-throughput analysis system for the interpretation of results from subtelomere assays will be of benefit to other Centres wishing to implement such an MLPA-based service.

  20. Ultraviolet-visible and fluorescence spectroscopy can be used as a diagnostic tool for gamma irradiation detection in vivo.

    Science.gov (United States)

    K-Abdelhalim, Mohamed Anwar; Moussa, Sherif A-Abdelmottaleb

    2016-09-01

    The spectroscopic properties can indicate important features about the nature and severity of the disease. However, no earlier studies have been used the spectroscopic properties as a diagnostic tool for radiation detection. This study was aimed to use ultraviolet-visible and fluorescence spectroscopy as a diagnostic tool for gamma irradiation detection in rats in vivo. Adult male rats were exposed to 25, 50, 75 and 100 Gray as single dose, using Cobalt-60 (Co-60) source with a dose rate of 0.883 centi Gray/sec (cGy/s). Ultraviolet and fluorescence spectroscopy of rat's blood serum were measured. After gamma irradiation of rats in vivo, the blood serum absorbance peaks for 25, 50, 75 and 100 Gray (Gy) decreased and shifted towards the ultra violet wavelength. A maximal change in fluorescence intensity of blood serum at 350 nm was obtained when exciting light at 194 nm after irradiation. The fluorescence intensity also decreased with the dose. The highest radiation gamma dose might be accompanied with the highest oxidative stress. This study suggests that at the above mentioned gamma radiation doses, the blood is highly fragmented; with low aggregation at 25 Gy and with high aggregation at 50-100 Gy.

  1. Evaluation of the efficiency of biofield diagnostic system in breast cancer detection using clinical study results and classifiers.

    Science.gov (United States)

    Subbhuraam, Vinitha Sree; Ng, E Y K; Kaw, G; Acharya U, Rajendra; Chong, B K

    2012-02-01

    The division of breast cancer cells results in regions of electrical depolarisation within the breast. These regions extend to the skin surface from where diagnostic information can be obtained through measurements of the skin surface electropotentials using sensors. This technique is used by the Biofield Diagnostic System (BDS) to detect the presence of malignancy. This paper evaluates the efficiency of BDS in breast cancer detection and also evaluates the use of classifiers for improving the accuracy of BDS. 182 women scheduled for either mammography or ultrasound or both tests participated in the BDS clinical study conducted at Tan Tock Seng hospital, Singapore. Using the BDS index obtained from the BDS examination and the level of suspicion score obtained from mammography/ultrasound results, the final BDS result was deciphered. BDS demonstrated high values for sensitivity (96.23%), specificity (93.80%), and accuracy (94.51%). Also, we have studied the performance of five supervised learning based classifiers (back propagation network, probabilistic neural network, linear discriminant analysis, support vector machines, and a fuzzy classifier), by feeding selected features from the collected dataset. The clinical study results show that BDS can help physicians to differentiate benign and malignant breast lesions, and thereby, aid in making better biopsy recommendations.

  2. Diagnostic performance of cone-beam computed tomography on detection of mechanically-created artificial secondary caries

    Energy Technology Data Exchange (ETDEWEB)

    Charuakkra, Arnon; Prapayasatok, Sangsom; Janhom, Apirum; Pongsirwet, Surawut; Verochana, Karune; Mahasantipiya, Phattaranant [Faculty of Dentistry, Chiang Mai University, Chiang Mai (Thailand)

    2011-12-15

    The aim of this study was to compare the diagnostic accuracy of cone-beam computed tomography (CBCT) images and bitewing images in detection of secondary caries. One hundred and twenty proximal slots of Class II cavities were randomly prepared on human premolar and molar teeth, and restored with amalgam (n=60) and composite resin (n=60). Then, artificial secondary caries lesions were randomly created using round steel No. 4 bur. The teeth were radiographed with a conventional bitewing technique and two CBCT systems; Pax-500ECT and Promax 3D. All images were evaluated by five observers. The area under the receiver operating characteristic (ROC) curve (Az) was used to evaluate the diagnostic accuracy. Significant difference was tested using the Friedman test (p value<0.05). The mean Az values for bitewing, Pax-500ECT, and Promax 3D imaging systems were 0.882, 0.995, and 0.978, respectively. Significant differences were found between the two CBCT systems and film (p=0.007). For CBCT systems, the axial plane showed the greatest Az value. Based on the design of this study, CBCT images were better than bitewing radiographs in detection of secondary caries.

  3. A Review on Traditional Caries Diagnostic Systems and Introduction of New International Caries Detection and Assessment System (ICDAS

    Directory of Open Access Journals (Sweden)

    Malek Mohammadi T.

    2011-04-01

    Full Text Available Dental caries is a chronic infection disease that manifests itself through demineralization of dental tissues and cavitations in the tooth structure. The diagnosis of dental caries poses challenges due to the complex interaction of multiple endogenous causal factors. Naturally, visual and visual-tactile examinations have been the most commonly used diagnostic aids in caries examination for many years. A major drawback in common dental examination was the use of varied diagnostic criteria by various authors. Some extensive reviews regarding this have emphasized the need for a unique system for the diagnosis of such caries. Thus, International Caries Detection and Assessment System (ICDAS emerged as a benchmark for detection and assessment of dental caries. The main objective of the new system is to improve the level of information to help make informed decision about diagnosis, prognosis, and clinical management of caries at both individual and public health levels.This study aimed to review traditional caries diagnosis methods and also describe the philosophy of the new ICDAS system for diagnosis and assessment of caries and also demonstrate how it can serve as a basis and benchmark for clinical and epidemiological research.

  4. Diagnostic accuracy of 64-slice multidetector CT for detection of in-stent restenosis in an unselected, consecutive patient population

    Energy Technology Data Exchange (ETDEWEB)

    Haraldsdottir, Sigurdis, E-mail: sigurdisha@gmail.com [Boston Medical Center, 72 East Concord Street (Evans 124), Boston, MA, 02118 (United States); Gudnason, Thorarinn, E-mail: thorgudn@landspitali.is [Landspitali University Hospital, Hringbraut, 101 Reykjavik (Iceland); Sigurdsson, Axel F., E-mail: axelfsig@landspitali.is [Landspitali University Hospital, Hringbraut, 101 Reykjavik (Iceland); Gudjonsdottir, Jonina, E-mail: jonina@rd.is [Rontgen Domus Medica, Egilsgata 3, 101 Reykjavik (Iceland); Lehman, Sam J., E-mail: slehman@partners.org [Massachusetts General Hospital, 165 Cambridge Street, Suite 400, Boston, MA 02114 (United States); Eyjolfsson, Kristjan, E-mail: kristey@landspitali.is [Landspitali University Hospital, Hringbraut, 101 Reykjavik (Iceland); Scheving, Sigurpall S., E-mail: sigurpal@landspitali.is [Landspitali University Hospital, Hringbraut, 101 Reykjavik (Iceland); Gibson, C. Michael, E-mail: mgibson@perfuse.org [Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, MA 02115 (United States); Hoffmann, Udo, E-mail: uhoffmann@partners.org [Massachusetts General Hospital, 165 Cambridge Street, Suite 400, Boston, MA 02114 (United States); Jonsdottir, Birna, E-mail: birna@rd.is [Rontgen Domus Medica, Egilsgata 3, 101 Reykjavik (Iceland); Andersen, Karl, E-mail: andersen@landspitali.is [Landspitali University Hospital, Hringbraut, 101 Reykjavik (Iceland)

    2010-11-15

    Objectives: To investigate the diagnostic accuracy of 64-slice multidetector computed tomography (64-CT) for detection of in-stent restenosis (ISR) in an unselected, consecutive patient population. Background: Detection of in-stent restenosis by cardiac CT would be a major advance for the evaluation of patients suspected of having ISR. However, the diagnostic accuracy of current generation 64-CT in this context is not fully established. Methods: We conducted a prospective study on patients with stable angina or acute coronary syndrome with no prior history of coronary artery disease. Six months after percutaneous coronary intervention (PCI) with stent placement they underwent a 64-CT scan (Toshiba Multi-Slice Aquilion 64) and consequently a repeat coronary angiography for comparison. Cardiac CT data sets were analyzed for the presence of in-stent restenosis by two independent expert readers blinded to the coronary angiographic data. Results: Ninety-three patients with a total of 140 stents were evaluated. Males comprised 82% of the study group and the mean age was 63 {+-} 10 years. The mean time from PCI to the repeat coronary angiography was 208 {+-} 37 days and the mean time from 64-CT to repeat coronary angiography was 3.7 {+-} 4.9 days. The restenosis rate according to coronary angiography was 26%. Stent diameter, strut thickness, heart rate and body mass index (BMI) significantly affected image quality. The sensitivity, specificity, positive and negative predictive values of 64-CT for detection of in-stent restenosis were 27%, 95%, 67% and 78%, respectively. Conclusions: Current generation, 64-slice CT, remains limited in its ability to accurately detect in-stent restenosis.

  5. An Electrochromatography Chip with Integrated Waveguides for UV Absorbance Detection

    DEFF Research Database (Denmark)

    Gustafsson, Omar; Mogensen, Klaus Bo; Ohlsson, Pelle Daniel

    2008-01-01

    A silicon-based microchip for electrochromatographic separations is presented. Apart from a microfluidic network, the microchip has integrated UV-transparent waveguides for detection and integrated couplers for optical fibers on the chip, yielding the most complete chromatography microchip to date...... in terms of the integration of optical components. The microfluidic network and the optical components are fabricated in a single etching step in silicon and subsequently thermally oxidized. The separation column consists of a regular array of microfabricated solid support structures with a monolayer...

  6. Self-transport and self-alignment of microchips using microscopic rain

    Science.gov (United States)

    Chang, Bo; Shah, Ali; Zhou, Quan; Ras, Robin H. A.; Hjort, Klas

    2015-10-01

    Alignment of microchips with receptors is an important process step in the construction of integrated micro- and nanosystems for emerging technologies, and facilitating alignment by spontaneous self-assembly processes is highly desired. Previously, capillary self-alignment of microchips driven by surface tension effects on patterned surfaces has been reported, where it was essential for microchips to have sufficient overlap with receptor sites. Here we demonstrate for the first time capillary self-transport and self-alignment of microchips, where microchips are initially placed outside the corresponding receptor sites and can be self-transported by capillary force to the receptor sites followed by self-alignment. The surface consists of hydrophilic silicon receptor sites surrounded by superhydrophobic black silicon. Rain-induced microscopic droplets are used to form the meniscus for the self-transport and self-alignment. The boundary conditions for the self-transport have been explored by modeling and confirmed experimentally. The maximum permitted gap between a microchip and a receptor site is determined by the volume of the liquid and by the wetting contrast between receptor site and substrate. Microscopic rain applied on hydrophilic-superhydrophobic patterned surfaces greatly improves the capability, reliability and error-tolerance of the process, avoiding the need for accurate initial placement of microchips, and thereby greatly simplifying the alignment process.

  7. Self-transport and self-alignment of microchips using microscopic rain.

    Science.gov (United States)

    Chang, Bo; Shah, Ali; Zhou, Quan; Ras, Robin H A; Hjort, Klas

    2015-10-09

    Alignment of microchips with receptors is an important process step in the construction of integrated micro- and nanosystems for emerging technologies, and facilitating alignment by spontaneous self-assembly processes is highly desired. Previously, capillary self-alignment of microchips driven by surface tension effects on patterned surfaces has been reported, where it was essential for microchips to have sufficient overlap with receptor sites. Here we demonstrate for the first time capillary self-transport and self-alignment of microchips, where microchips are initially placed outside the corresponding receptor sites and can be self-transported by capillary force to the receptor sites followed by self-alignment. The surface consists of hydrophilic silicon receptor sites surrounded by superhydrophobic black silicon. Rain-induced microscopic droplets are used to form the meniscus for the self-transport and self-alignment. The boundary conditions for the self-transport have been explored by modeling and confirmed experimentally. The maximum permitted gap between a microchip and a receptor site is determined by the volume of the liquid and by the wetting contrast between receptor site and substrate. Microscopic rain applied on hydrophilic-superhydrophobic patterned surfaces greatly improves the capability, reliability and error-tolerance of the process, avoiding the need for accurate initial placement of microchips, and thereby greatly simplifying the alignment process.

  8. Physiological and behavioural responses of young horses to hot iron branding and microchip implantation.

    Science.gov (United States)

    Erber, R; Wulf, M; Becker-Birck, M; Kaps, S; Aurich, J E; Möstl, E; Aurich, C

    2012-02-01

    Branding is the traditional and well-established method used to mark horses, but recently microchip transponders for implantation have become available. In this study, behaviour, physiological stress variables and skin temperature in foals were determined in response to hot-iron branding (n=7) and microchip implantation (n=7). Salivary cortisol concentrations increased in response to branding (1.8 ± 0.2 ng/mL) and microchip implantation (1.4 ± 0.1ng/mL), but cortisol release over time did not differ. In response to both manipulations there was a transient increase in heart rate (PBranding and microchip implantation induced a comparable aversive behaviour (branding, score 3.86 ± 0.85; microchip, score 4.00 ± 0.82). Both techniques thus caused similar physiological and behavioural changes indicative of stress. Acutely, implantation of a microchip was as stressful as branding in foals. Branding caused a necrotising skin burn lasting at least 7 days. Moreover branding, but not microchip implantation (P<0.001), was accompanied by a generalized increase in skin temperature which was comparable to low degree post-burn hypermetabolism in humans. Copyright © 2011 Elsevier Ltd. All rights reserved.

  9. The Scientific Method, Diagnostic Bayes, and How to Detect Epistemic Errors

    Science.gov (United States)

    Vrugt, J. A.

    2015-12-01

    In the past decades, Bayesian methods have found widespread application and use in environmental systems modeling. Bayes theorem states that the posterior probability, P(H|D) of a hypothesis, H is proportional to the product of the prior probability, P(H) of this hypothesis and the likelihood, L(H|hat{D}) of the same hypothesis given the new/incoming observations, \\hat {D}. In science and engineering, H often constitutes some numerical simulation model, D = F(x,.) which summarizes using algebraic, empirical, and differential equations, state variables and fluxes, all our theoretical and/or practical knowledge of the system of interest, and x are the d unknown parameters which are subject to inference using some data, \\hat {D} of the observed system response. The Bayesian approach is intimately related to the scientific method and uses an iterative cycle of hypothesis formulation (model), experimentation and data collection, and theory/hypothesis refinement to elucidate the rules that govern the natural world. Unfortunately, model refinement has proven to be very difficult in large part because of the poor diagnostic power of residual based likelihood functions tep{gupta2008}. This has inspired te{vrugt2013} to advocate the use of 'likelihood-free' inference using approximate Bayesian computation (ABC). This approach uses one or more summary statistics, S(\\hat {D}) of the original data, \\hat {D} designed ideally to be sensitive only to one particular process in the model. Any mismatch between the observed and simulated summary metrics is then easily linked to a specific model component. A recurrent issue with the application of ABC is self-sufficiency of the summary statistics. In theory, S(.) should contain as much information as the original data itself, yet complex systems rarely admit sufficient statistics. In this article, we propose to combine the ideas of ABC and regular Bayesian inference to guarantee that no information is lost in diagnostic model

  10. The microbial detection array for detection of emerging viruses in clinical samples--a useful panmicrobial diagnostic tool

    DEFF Research Database (Denmark)

    Rosenstierne, Maiken W; McLoughlin, Kevin S; Olesen, Majken Lindholm

    2014-01-01

    emerging viruses or common infections, making these unexpected pathogens difficult to diagnose. Broad-spectrum pathogen detection microarrays containing probes for all sequenced viruses and bacteria can provide rapid identification of viruses, guiding decisions about treatment and appropriate case......Emerging viruses are usually endemic to tropical and sub-tropical regions of the world, but increased global travel, climate change and changes in lifestyle are believed to contribute to the spread of these viruses into new regions. Many of these viruses cause similar disease symptoms as other...... management. We report a modified Whole Transcriptome Amplification (WTA) method that increases unbiased amplification, particular of RNA viruses. Using this modified WTA method, we tested the specificity and sensitivity of the Lawrence Livermore Microbial Detection Array (LLMDA) against a wide range...

  11. COMPARISON OF DIAGNOSTIC METHODS FOR DETECTING AN INFLUENTIAL OBSERVATION IN REGRESSION

    OpenAIRE

    ACARLAR, Irmak

    2011-01-01

    An influential observation and influential sets would cause noticeable differentiations on the fitted values in regression. Since these differentiations decrease explicable of model, detecting the influential observation or the influential sets in data is important for efficiency of regression analysis. In this study DFFITS, DFBETAS, COVRATIO, Cook Distance, S statistics and graphical technique used for detecting an influential observation are examined. These methods are compared with regard ...

  12. Resorbable polymer microchips releasing BCNU inhibit tumor growth in the rat 9L flank model.

    Science.gov (United States)

    Kim, Grace Y; Tyler, Betty M; Tupper, Malinda M; Karp, Jeffrey M; Langer, Robert S; Brem, Henry; Cima, Michael J

    2007-11-01

    Sustained local delivery of single agents and controlled delivery of multiple chemotherapeutic agents are sought for the treatment of brain cancer. A resorbable, multi-reservoir polymer microchip drug delivery system has been tested against a tumor model. The microchip reservoirs were loaded with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). BCNU was more stable at 37 degrees C within the microchip compared to a uniformly impregnated polymeric wafer (70% intact drug vs. 38%, at 48 h). The half-life of the intact free drug in the microchip was 11 days, which is a marked enhancement compared to its half-life in normal saline and 10% ethanol (7 and 10 min, respectively) [P. Tepe, S.J. Hassenbusch, R. Benoit, J.H. Anderson, BCNU stability as a function of ethanol concentration and temperature, J. Neurooncol. 10 (1991) 121-127; P. Kari, W.R. McConnell, J.M. Finkel, D.L. Hill, Distribution of Bratton-Marshall-positive material in mice following intravenous injections of nitrosoureas, Cancer Chemother. Pharmacol. 4 (1980) 243-248]. A syngeneic Fischer 344 9L gliosarcoma rat model was used to study the tumoricidal efficacy of BCNU delivery from the microchip or homogeneous polymer wafer. A dose-dependent decrease in tumor size was found for 0.17, 0.67, and 1.24 mg BCNU-microchips. Tumors treated with 1.24 mg BCNU-microchips showed significant tumor reduction (p=0.001) compared to empty control microchips at two weeks. The treatment showed similar efficacy to a polymer wafer with the same dosage. The microchip reservoir array may enable delivery of multiple drugs with independent release kinetics and formulations.

  13. Photon undulatory non-linear conversion diagnostic method for caries detection: a pilot study.

    Science.gov (United States)

    Kesler, G; Masychev, V; Sokolovsky, A; Alexandrov, M; Kesler, A; Koren, R

    2003-08-01

    The objective of this study was to evaluate a new optical method - photon undulatory non-linear conversion (PNC)--for use in different stages of caries detection. Caries should be considered an infectious disease managed by risk assessment, early detection, and preventive therapies, rather than simply "drilling and filling." Fluorescence emission spectroscopy was performed in vitro on 90 extracted teeth, with intact occlusal surfaces. This system differs from the basic Diagnodent unit in its ability to distinguish between different tissue components with respect to their spectrums. Histological analysis served as the gold standard for verification. The teeth sections correspond to the specific point with the highest reading of the detector. The system was compared to visual inspection, probing, and x-ray methods. The system tested (helium-neon [He-Ne], lambda = 633 nm) has a fiber optic device that delivers radiation to the tooth and a spectrophotometer device that detects bacterial porphyrins fluorescence, allowing detection of caries, fillings, and calculus by simultaneous measurement of backscattering and fluorescence intensity. The system tested provides quantitatively reproducible measurements and detection even through sound enamel of more than 1 mm in thickness. The PNC method detects different stages of caries lesions in real time, and it exceeds x-rays in sensitivity, without any ionizing radiation. Preliminary results showed a high potential of using the PNC method in clinical practice (98% accuracy) in comparison to the other methods.

  14. The diagnostic value of lymph node biopsy to detect Castleman’s disease

    Directory of Open Access Journals (Sweden)

    Prashilla Soma

    2014-09-01

    Full Text Available HIV is not indicated in the aetiology of Castleman’s disease. However, it impacts on the prevalence and natural history of this disease and significantly on the disease progression. Castleman’s disease is a uni- or multicentric disease of the lymph node with or without polyclonal proliferation of B-cells. It is a morphologically distinct form of lymph node hyperplasia and is characterised by significant architectural changes in all lymphatic compartments. Histopathologically, the disease is classified into two major subtypes: the hyaline-vascular type and the plasma-cell type. A mixed type is also identified, as there are frequent transitions between the types. The diagnosis of Castleman’s disease needs to be made histologically. Treatment modalities include surgery, which is curative for unicentric disease, and systemic therapy, which is needed for multicentric disease. This case highlights the diagnostic value of lymph node excision biopsy in HIV-infected patients. 

  15. Accuracy and standardization of diagnostic methods for the detection of antibodies to citrullinated peptides

    Directory of Open Access Journals (Sweden)

    M. Tampoia

    2011-06-01

    Full Text Available Anti-citrullinated peptide antibodies (ACPA have a very high specificity for rheumatoid arthritis, much more than that of the rheumatoid factor. In addition, ACPA can be found in sera in the pre-clinical phase, are associated with more severe joint destruction and with higher disease activity. In recent years, keeping pace with new knowledge and with progress made in the antigenic composition of tests and in the characterization of immunogenic epitopes, many immunoenzymatic (ELISA methods of second and third generation have been produced and marketed commercially, and their use has spread among clinical laboratories. Today, completely automated methods are also available, which are easy to use and with a higher throughput, rendering the diagnostic utility of testing ever faster and more effective. This review takes into consideration the more important characteristics of the new ACPA-ELISA tests now commercially available, and also considers recent progress in standardizing test results.

  16. Diagnostic use of PCR for detection of Pneumocystis carinii in oral wash samples

    DEFF Research Database (Denmark)

    Helweg-Larsen, J; Jensen, Jens Ulrik Stæhr; Benfield, T

    1998-01-01

    and was compared to a previously described PCR protocol (mitochondrial RNA) run in a research laboratory. Both PCR methods amplified a sequence of the mitochondrial rRNA gene of P. carinii. Paired bronchoalveolar lavage (BAL) and oral wash specimens from 76 consecutive human immunodeficiency virus type 1-infected...... persons undergoing a diagnostic bronchoscopy were included. The TD-PCR procedure was quicker than the mitochondrial PCR procedure (compared to microscopy, had sensitivity, specificity, and positive and negative predictive values of 89, 94, 93, and 91%, respectively, for oral wash......There is a need to develop noninvasive methods for the diagnosis of Pneumocystis carinii pneumonia in patients unable to undergo bronchoscopy or induction sputum. Oral wash specimens are easily obtained, and P. carinii nucleic acid can be amplified and demonstrated by PCR. In routine clinical use...

  17. Limited diagnostic accuracy of magnetic resonance imaging and clinical tests for detecting partial-thickness tears of the rotator cuff.

    Science.gov (United States)

    Brockmeyer, Matthias; Schmitt, Cornelia; Haupert, Alexander; Kohn, Dieter; Lorbach, Olaf

    2017-09-23

    The reliable diagnosis of partial-thickness tears of the rotator cuff is still elusive in clinical practise. Therefore, the purpose of the study was to determine the diagnostic accuracy of MR imaging and clinical tests for detecting partial-thickness tears of the rotator cuff as well as the combination of these parameters. 334 consecutive shoulder arthroscopies for rotator cuff pathologies performed during the time period between 2010 and 2012 were analyzed retrospectively for the findings of common clinical signs for rotator cuff lesions and preoperative MR imaging. These were compared with the intraoperative arthroscopic findings as "gold standard". The reports of the MR imaging were evaluated with regard to the integrity of the rotator cuff. The Ellman Classification was used to define partial-thickness tears of the rotator cuff in accordance with the arthroscopic findings. Descriptive statistics, sensitivity, specificity, positive and negative predictive value were calculated. MR imaging showed 80 partial-thickness and 70 full-thickness tears of the rotator cuff. The arthroscopic examination confirmed 64 partial-thickness tears of which 52 needed debridement or refixation of the rotator cuff. Sensitivity for MR imaging to identify partial-thickness tears was 51.6%, specificity 77.2%, positive predictive value 41.3% and negative predictive value 83.7%. For the Jobe-test, sensitivity was 64.1%, specificity 43.2%, positive predictive value 25.9% and negative predictive value 79.5%. Sensitivity for the Impingement-sign was 76.7%, specificity 46.6%, positive predictive value 30.8% and negative predictive value 86.5%. For the combination of MR imaging, Jobe-test and Impingement-sign sensitivity was 46.9%, specificity 85.4%, positive predictive value 50% and negative predictive value 83.8%. The diagnostic accuracy of MR imaging and clinical tests (Jobe-test and Impingement-sign) alone is limited for detecting partial-thickness tears of the rotator cuff. Additionally

  18. Stand-Sit Microchip for High-Throughput, Multiplexed Analysis of Single Cancer Cells.

    Science.gov (United States)

    Ramirez, Lisa; Herschkowitz, Jason I; Wang, Jun

    2016-01-01

    Cellular heterogeneity in function and response to therapeutics has been a major challenge in cancer treatment. The complex nature of tumor systems calls for the development of advanced multiplexed single-cell tools that can address the heterogeneity issue. However, to date such tools are only available in a laboratory setting and don't have the portability to meet the needs in point-of-care cancer diagnostics. Towards that application, we have developed a portable single-cell system that is comprised of a microchip and an adjustable clamp, so on-chip operation only needs pipetting and adjusting of clamping force. Up to 10 proteins can be quantitated from each cell with hundreds of single-cell assays performed in parallel from one chip operation. We validated the technology and analyzed the oncogenic signatures of cancer stem cells by quantitating both aldehyde dehydrogenase (ALDH) activities and 5 signaling proteins in single MDA-MB-231 breast cancer cells. The technology has also been used to investigate the PI3K pathway activities of brain cancer cells expressing mutant epidermal growth factor receptor (EGFR) after drug intervention targeting EGFR signaling. Our portable single-cell system will potentially have broad application in the preclinical and clinical settings for cancer diagnosis in the future.

  19. Stand-Sit Microchip for High-Throughput, Multiplexed Analysis of Single Cancer Cells

    Science.gov (United States)

    Ramirez, Lisa; Herschkowitz, Jason I.; Wang, Jun

    2016-01-01

    Cellular heterogeneity in function and response to therapeutics has been a major challenge in cancer treatment. The complex nature of tumor systems calls for the development of advanced multiplexed single-cell tools that can address the heterogeneity issue. However, to date such tools are only available in a laboratory setting and don’t have the portability to meet the needs in point-of-care cancer diagnostics. Towards that application, we have developed a portable single-cell system that is comprised of a microchip and an adjustable clamp, so on-chip operation only needs pipetting and adjusting of clamping force. Up to 10 proteins can be quantitated from each cell with hundreds of single-cell assays performed in parallel from one chip operation. We validated the technology and analyzed the oncogenic signatures of cancer stem cells by quantitating both aldehyde dehydrogenase (ALDH) activities and 5 signaling proteins in single MDA-MB-231 breast cancer cells. The technology has also been used to investigate the PI3K pathway activities of brain cancer cells expressing mutant epidermal growth factor receptor (EGFR) after drug intervention targeting EGFR signaling. Our portable single-cell system will potentially have broad application in the preclinical and clinical settings for cancer diagnosis in the future. PMID:27581736

  20. Capillary Self-Alignment of Microchips on Soft Substrates

    Directory of Open Access Journals (Sweden)

    Bo Chang

    2016-03-01

    Full Text Available Soft micro devices and stretchable electronics have attracted great interest for their potential applications in sensory skins and wearable bio-integrated devices. One of the most important steps in building printed circuits is the alignment of assembled micro objects. Previously, the capillary self-alignment of microchips driven by surface tension effects has been shown to be able to achieve high-throughput and high-precision in the integration of micro parts on rigid hydrophilic/superhydrophobic patterned surfaces. In this paper, the self-alignment of microchips on a patterned soft and stretchable substrate, which consists of hydrophilic pads surrounded by a superhydrophobic polydimethylsiloxane (PDMS background, is demonstrated for the first time. A simple process has been developed for making superhydrophobic soft surface by replicating nanostructures of black silicon onto a PDMS surface. Different kinds of PDMS have been investigated, and the parameters for fabricating superhydrophobic PDMS have been optimized. A self-alignment strategy has been proposed that can result in reliable self-alignment on a soft PDMS substrate. Our results show that capillary self-alignment has great potential for building soft printed circuits.

  1. Energy Conversion from Salinity Gradient Using Microchip with Nafion Membrane

    Science.gov (United States)

    Chang, Che-Rong; Yeh, Ching-Hua; Yeh, Hung-Chun; Yang, Ruey-Jen

    2016-06-01

    When a concentrated salt solution and a diluted salt solution are separated by an ion-selective membrane, cations and anions would diffuse at different rates depending on the ion selectivity of the membrane. The difference of positive and negative charges at both ends of the membrane would produce a potential, called the diffusion potential. Thus, electrical energy can be converted from the diffusion potential through reverse electrodialysis. This study demonstrated the fabrication of an energy conversion microchip using the standard micro-electromechanical technique, and utilizing Nafion junction as connecting membrane, which was fabricated by a surface patterned process. Through different salinity gradient of potassium chloride solutions, we experimentally investigated the diffusion potential and power generation from the microchip, and the highest value measured was 135 mV and 339 pW, respectively. Furthermore, when the electrolyte was in pH value of 3.8, 5.6, 10.3, the system exhibited best performance at pH value of 10.3; whereas, pH value of 3.8 yielded the worst.

  2. Diagnostic approach for cancer cells in urine sediments by 5-aminolevulinic acid-based photodynamic detection in bladder cancer.

    Science.gov (United States)

    Miyake, Makito; Nakai, Yasushi; Anai, Satoshi; Tatsumi, Yoshihiro; Kuwada, Masaomi; Onishi, Sayuri; Chihara, Yoshitomo; Tanaka, Nobumichi; Hirao, Yoshihiko; Fujimoto, Kiyohide

    2014-05-01

    Bladder urothelial carcinoma is diagnosed and followed up after transurethral resection using a combination of cystoscopy, urine cytology and urine biomarkers at regular intervals. However, cystoscopy can overlook flat lesions like carcinoma in situ, and the sensitivity of urinary tests is poor in low-grade tumors. There is an emergent need for an objective and easy urinary diagnostic test for the management of bladder cancer. In this study, three different modalities for 5-aminolevulinic acid (ALA)-based photodynamic diagnostic tests were used. We developed a compact-size, desktop-type device quantifying red fluorescence in cell suspensions, named "Cellular Fluorescence Analysis Unit" (CFAU). Urine samples from 58 patients with bladder cancer were centrifuged, and urine sediments were then treated with ALA. ALA-treated sediments were subjected to three fluorescence detection assays, including the CFAU assay. The overall sensitivities of conventional cytology, BTA, NMP22, fluorescence cytology, fluorescent spectrophotometric assay and CFAU assay were 48%, 33%, 40%, 86%, 86% and 87%, respectively. Three different ALA-based assays showed high sensitivity and specificity. The ALA-based assay detected low-grade and low-stage bladder urothelial cells at shigher rate (68-80% sensitivity) than conventional urine cytology, BTA and NMP22 (8-20% sensitivity). Our findings demonstrate that the ALA-based fluorescence detection assay is promising tool for the management of bladder cancer. Development of a rapid and automated device for ALA-based photodynamic assay is necessary to avoid the variability induced by troublesome steps and low stability of specimens.

  3. Time-delayed contrast-enhanced MRI improves detection of brain metastases: a prospective validation of diagnostic yield.

    Science.gov (United States)

    Cohen-Inbar, Or; Xu, Zhiyuan; Dodson, Blair; Rizvi, Tanvir; Durst, Christopher R; Mukherjee, Sugoto; Sheehan, Jason P

    2016-12-01

    The radiological detection of brain metastases (BMs) is essential for optimizing a patient's treatment. This statement is even more valid when stereotactic radiosurgery, a noninvasive image guided treatment that can target BM as small as 1-2 mm, is delivered as part of that care. The timing of image acquisition after contrast administration can influence the diagnostic sensitivity of contrast enhanced magnetic resonance imaging (MRI) for BM. Investigate the effect of time delayed acquisition after administration of intravenous Gadavist® (Gadobutrol 1 mmol/ml) on the detection of BM. This is a prospective IRB approved study of 50 patients with BM who underwent post-contrast MRI sequences after injection of 0.1 mmol/kg Gadavist® as part of clinical care (time-t0), followed by axial T1 sequences after a 10 min (time-t1) and 20 min delay (time-t2). MRI studies were blindly compared by three neuroradiologists. Single measure intraclass correlation coefficients were very high (0.914, 0.904 and 0.905 for time-t0, time-t1 and time-t2 respectively), corresponding to a reliable inter-observer correlation. The delayed MRI at time-t2 delayed sequences showed a significant and consistently higher diagnostic sensitivity for BM by every participating neuroradiologist and for the entire cohort (p = 0.016, 0.035 and 0.034 respectively). A disproportionately high representation of BM detected on the delayed studies was located within posterior circulation territories (compared to predictions based on tissue volume and blood-flow volumes). Considering the safe and potentially high yield nature of delayed MRI sequences, it should supplement the standard MRI sequences in all patients in need of precise delineation of their intracranial disease.

  4. Comparison of diagnostic value of multidetector computed tomography and X-ray in the detection of body packing

    Energy Technology Data Exchange (ETDEWEB)

    Bulakci, Mesut, E-mail: mesutbulakci@yahoo.com [Department of Radiology, Haseki Training and Research Hospital, 34096 Aksaray, Istanbul (Turkey); Kalelioglu, Tuba, E-mail: tubakarsakarya@hotmail.com [Department of Radiology, Haseki Training and Research Hospital, 34096 Aksaray, Istanbul (Turkey); Bulakci, Betul Bozkurt, E-mail: dr.betulbozkurt@gmail.com [Department of Family Medicine, Istanbul University, Istanbul Faculty of Medicine, 34390 Capa, Istanbul (Turkey); Kiris, Adem, E-mail: ademkiris@hotmail.com [Department of Radiology, Haseki Training and Research Hospital, 34096 Aksaray, Istanbul (Turkey)

    2013-08-15

    Objective: Radiologists and other clinicians are facing an increasing number of illegal drug-related medical conditions. We aimed to draw attention to this growing global problem and to highlight some of the important points related to diagnosis and follow-up of body packing. We compare the diagnostic performance of unenhanced multidetector CT (MDCT) and abdomen X-ray for the detection of drug-filled packets. Materials and methods: Sixty-seven suspects, who underwent both CT and X-ray examinations, have been included in the study. All MDCT and X-ray images were independently and retrospectively reviewed by two observers with different degrees of experience in abdomen imaging. Fifty-two of them were identified as body packers finally. Interobserver agreement, sensitivity, specificity, positive and negative predictive value were calculated. Results: Two types of packets with different characteristics were identified in all body packers. Type 1 packets (solid-state drug) were found in 41 patients and type 2 packets (liquid cocaine) in 11 patients. All statistical analyses concern the detection of any packets. That is, the whole evaluation has been performed per patient. Sensitivity/specificity values of type 1 and type 2 packets for MDCT were 100–98%/100–100% and 100–100%/100–100%, respectively. Besides, sensitivity/specificity values of type 1 and type 2 packets for X-ray were 93–90%/100–91% and 64–45%/73–71%, respectively. In addition, interobserver agreements for detection of any packets were excellent (κ = 0.96) and good (κ = 0.75) for interpretation of MDCT and X-ray, respectively. Conclusion: Unenhanced MDCT is a fast, accurate and easily used diagnostic tool with high sensitivity and specificity for the exact diagnosis of body packing.

  5. Reduced-size microchips for identification of horses: response to implantation and readability during a six-month period.

    Science.gov (United States)

    Wulf, M; Aurich, C; von Lewinski, M; Möstl, E; Aurich, J E

    2013-11-09

    In this study, readability of reduced-size microchips in horses and the response to implantation were analysed. It was hypothesised that small microchips can be implanted stress-free but are less readable than larger microchips. Adult mares (n=40) were implanted with a reduced-size microchip (10.9×1.6 mm) at the left side of the neck (size of conventional microchips 11.4×2.2 mm). Microchips were identified with three different scanners (A, B, C) immediately, and at 6, 12 and 28 weeks after implantation. Twelve out of the 40 mares were submitted to microchip implantation and control treatments and cortisol, heart rate and heart rate variability (HRV) were determined. From the chip-bearing side of the neck, microchips were identified with all scanners in all horses at all times. From the contralateral side, correct readings were always 100 per cent with scanner C and with scanners A and B ranged between 60 and 100 per cent. Heart rate and HRV variable sd of beat-to-beat interval increased slightly (Phorses. Compared with conventional microchips, the reduction in size did not impair readability. Microchip implantation is no pronounced stressor for horses.

  6. Biomarkers for Early Detection of Malignant Mesothelioma: Diagnostic and Therapeutic Application

    Directory of Open Access Journals (Sweden)

    Marco Tomasetti

    2010-04-01

    Full Text Available Malignant mesothelioma (MM is a rare and aggressive tumour of the serosal cavities linked to asbestos exposure. Improved detection methods for diagnosing this type of neoplastic disease are essential for an early and reliable diagnosis and treatment. Thus, focus has been placed on finding tumour markers for the non-invasive detection of MM. Recently, some blood biomarkers have been described as potential indicators of early and advanced MM cancers. The identification of tumour biomarkers alone or in combination could greatly facilitate the surveillance procedure for cohorts of subjects exposed to asbestos, a common phenomenon in several areas of western countries.

  7. The diagnostic potential of maternal plasma in detecting fetal diseases by DNA test

    Directory of Open Access Journals (Sweden)

    Saha Biswajit

    2004-01-01

    Full Text Available Conventionally, DNA based investigations for fetal diseases are done by chorionic villous sampling and amniocentesis. Both are invasive techniques. Recently, molecular diagnosis has also been made possible in early pregnancy from maternal blood which is noninvasive and advantageous. Most of the researches have tried to identify the Y chromosome marker(s to detect a male fetus and paternally inherited allele. This is currently helpful to detect a very few genetic disorders including Rh D status in Rh negative women in early pregnancy and preeclampsia a few weeks preceding the clinical onset. This is a potential area for prenatal diagnosis in future.

  8. Partial shadowing detection based on equivalent thermal voltage monitoring for PV module diagnostics

    DEFF Research Database (Denmark)

    Sera, Dezso; Teodorescu, Remus; Rodriguez, Pedro

    2009-01-01

    Partial shadowing of photovoltaic systems can overproportionally reduce the energy yield and lead to early ageing and failure of the shadowed cells. Large area shadows are relatively easy to detect due to the eminent power reduction and decrease of fill factor. However, small area partial shadows...... in a larger system do not have a very obvious effect on the output power or fill factor of the PV array and can remain undetected, leading to failure. In this paper a method for detecting small area partial shadows, based on equivalent thermal voltage, is presented. A simplified expression of the equivalent...

  9. Papanicolau smear chances to be diagnostic for cervical squamous intraepithelial lesions (SIL) with or without detectable HPV DNA at in situ hybridization analysis.

    Science.gov (United States)

    Sopracordevole, F; Cadorin, L; Muffato, G; De Benetti, L; Parin, A

    1993-01-01

    The Authors have correlated 39 cervical diagnostic biopsies for squamous intraepithelial lesions (SILs) with correspective Papanicolau smears (PS), with relation to the presence or the absence of HPV of oncogenic type (HPV-one) detected by in situ hybridization (ISH). Agreement between cytological and histological diagnosis was present in 14 of 16 cases with detectable HPV-one and only in 12 of 23 cases without detectable HPV-one at ISH. The importance of the HPV type in the SILs with relation to the diagnostic accuracy of Papanicolaou smears has been discussed.

  10. Diagnostic performance of fecal quantitative real-time polymerase chain reaction for detection of Lawsonia intracellularis–associated proliferative enteropathy in nursery pigs

    DEFF Research Database (Denmark)

    Pedersen, Ken Steen; Stege, Helle; Jensen, Tim Kåre;

    2013-01-01

    Quantitative polymerase chain reaction (qPCR) tests for detection and quantification of Lawsonia intracellularis in feces from pigs have been developed. The objective of the current study was to evaluate the diagnostic performance of a fecal qPCR test for detection of nursery pigs with L...

  11. A Novel Technique for Detecting Antibiotic-Resistant Typhoid from Rapid Diagnostic Tests

    OpenAIRE

    2015-01-01

    Fluoroquinolone-resistant typhoid is increasing. An antigen-detecting rapid diagnotic test (RDT) can rapidly diagnose typhoid from blood cultures. A simple, inexpensive molecular technique performed with DNA from positive RDTs accurately identified gyrA mutations consistent with phenotypic susceptibility testing results. Field diagnosis combined with centralized molecular resistance testing could improve typhoid management and surveillance in low-resource settings.

  12. Positive diagnostic values and histological detection ratios from the Rotterdam cervical cancer screening programme

    NARCIS (Netherlands)

    F.A. Kreuger; H. Beerman (Henk); H.G. Nijs (Huub); M. van Ballegooijen (Marjolein)

    1998-01-01

    textabstractBACKGROUND: In organized screening programmes for cervical cancer, pre-cancerous lesions are detected by cervical smears. However, during follow-up after a positive smear these pre-cancerous lesions are not always found. The purpose of the study is to analys

  13. Positive diagnostic values and histological detection ratios from the Rotterdam cervical cancer screening programme

    NARCIS (Netherlands)

    F.A. Kreuger; H. Beerman (Henk); H.G. Nijs (Huub); M. van Ballegooijen (Marjolein)

    1998-01-01

    textabstractBACKGROUND: In organized screening programmes for cervical cancer, pre-cancerous lesions are detected by cervical smears. However, during follow-up after a positive smear these pre-cancerous lesions are not always found. The purpose of the study is to analys

  14. Anti-cysticercus antibody detection in saliva as a potential diagnostic tool for neurocysticercosis

    Science.gov (United States)

    Saha, Rumpa; Roy, Priyamvada; Das, Shukla; Shah, Dheeraj; Agarwal, Sunil; Kaur, Iqbal Rajinder

    2016-01-01

    Objectives: This study was planned to determine the usefulness of anti-cysticercus IgG antibody detection in saliva for neurocysticercosis (NCC) diagnosis, along with serum C-reactive protein (CRP) level to serve as a surrogate marker. Materials and Methods: In this prospective study of 14 months duration, blood and saliva samples were collected from 40 patients suspected to be suffering from NCC and were subjected to anti-cysticercus IgG antibody detection by ELISA. Serum CRP levels were estimated as acute-phase reactant by high sensitivity CRP ELISA. Results: Anti-cysticercus IgG was detected in serum and saliva of 34 and 30 patients, respectively. Cases positive for salivary antibody were positive for serum antibody and their serum CRP level was higher than normal. Cases negative for salivary antibody had low serum CRP levels. Anti-cysticercus IgG detection in saliva was 88.24% sensitive, 100% specific, and had a positive predictive value of 100% and negative predictive value of 60%. Positive salivary anti-cysticercus IgG and high serum CRP level showed a significant association. Difference between CRP levels of patients positive for anti-cysticercus antibody in both serum and saliva, and patients positive for antibody in serum but not saliva was highly significant. Conclusions: Saliva, being painless and noninvasive, can be used as alternative to serum for NCC diagnosis. PMID:27570404

  15. Human Plasmodium knowlesi infection detected by rapid diagnostic tests for malaria

    NARCIS (Netherlands)

    J.J. van Hellemond (Jaap); M. Rutten (Martine); R. Koelewijn (Rob); A.M. Zeeman (Anne Marie); J. Verweij (Jaap); P.J. Wismans (Pieter); C.H. Kocken (Clemens); P.J.J. van Genderen (Perry)

    2009-01-01

    textabstractWe describe a PCR-confirmed case of Plasmodium knowlesi infection with a high parasitemia level and clinical signs of severe malaria in a migrant worker from Malaysian Borneo in the Netherlands. Investigations showed that commercially available rapid antigen tests for detection of human

  16. Population Based Screening for Prostate Cancer: assessment of diagnostic tools and cancers detected

    NARCIS (Netherlands)

    J.B.W. Rietbergen (John)

    1998-01-01

    textabstractOver the past decade, considerable debate has occurred over the question whether or not to screen asymptomatic men for prostate cancer. It is unknown whether early detection and treatment of the disease will decrease the disease specific mortality. On theoretical grounds screening may pr

  17. The Diagnostic Accuracy of Screening Tools to Detect Eating Disorders in Female Athletes.

    Science.gov (United States)

    Wagner, Alyssa J; Erickson, Casey D; Tierney, Dayna K; Houston, Megan N; Bacon, Cailee E Welch

    2016-12-01

    Clinical Scenario: Eating disorders in female athletes are a commonly underdiagnosed condition. Better screening tools for eating disorders in athletic females could help increase diagnosis and help athletes get the treatment they need. Focused Clinical Question: Should screening tools be used to detect eating disorders in female athletes? Summary of Key Findings: The literature was searched for studies that included information regarding the sensitivity and specificity of screening tools for eating disorders in female athletes. The search returned 5 possible articles related to the clinical question; 3 studies met the inclusion criteria (2 cross-sectional studies, 1 cohort study) and were included. All 3 studies reported sensitivity and specificity for the Athletic Milieu Direct Questionnaire version 2, the Brief Eating Disorder in Athletes Questionnaire version 2, and the Physiologic Screening Test to Detect Eating Disorders Among Female Athletes. All 3 studies found that the respective screening tool was able to accurately identify female athletes with eating disorders; however, the screening tools varied in sensitivity and specificity values. Clinical Bottom Line: There is strong evidence to support the use of screening tools to detect eating disorders in female athletes. Screening tools with higher sensitivity and specificity have demonstrated a successful outcome of determining athletes with eating disorders or at risk for developing an eating disorder. Strength of Recommendation: There is grade A evidence available to demonstrate that screening tools accurately detect female athletes at risk for eating disorders.

  18. Rapid, whole blood diagnostic test for detecting anti-hantavirus antibody in rats.

    Science.gov (United States)

    Amada, Takako; Yoshimatsu, Kumiko; Yasuda, Shumpei P; Shimizu, Kenta; Koma, Takaaki; Hayashimoto, Nobuhito; Gamage, Chandika D; Nishio, Sanae; Takakura, Akira; Arikawa, Jiro

    2013-10-01

    Hantavirus is a causative agent of rodent-borne viral zoonoses, hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome. Seoul virus (SEOV) is a causative agent of urban and laboratory rat-associated HFRS worldwide. Surveillance of rodents has been done mainly by serological detection of hantavirus-specific antibodies by enzyme linked immunosorbent assay (ELISA) and immunofluorescent antibody assay (IFA). An immunochromatographic (ICG) test was developed with the N-terminal 103 amino acids of nucleocapsid protein of Hantaan virus expressed by Escherichia coli as an antigen to detect IgG antibody specific to hantavirus in sera from Rattus sp. animals. Antibody-detecting sensitivity of the ICG test was the same as that of ELISA and about 100-times higher than that of IFA. Overall sensitivities and specificities of the ICG test in comparison to ELISA and IFA for sera from 192 urban rats and 123 laboratory rats were 99.3% and 100%, respectively. Diluted whole blood samples without separation could be used for the ICG test. The ICG test enabled detection of antibodies to SEOV, Hantaan, Dobrava/Belgrade, and Thailand viruses, which are causative agents of HFRS throughout Eurasia. The ICG test is a rapid, simple and safe method for diagnosis of SEOV infection in rats.

  19. Novel protease-based diagnostic devices for detection of wound infection

    NARCIS (Netherlands)

    Heinzle, A.; Papen-Botterhuis, N.E.; Schiffer, D.; Schneider, K.P.; Binder, B.; Schintler, M.; Haaksman, I.K.; Lenting, H.B.; Gübitz, G.M.; Sigl, E.

    2013-01-01

    A gelatinase-based device for fast detection of wound infection was developed. Collective gelatinolytic activity in infected wounds was 23 times higher (p ≤ 0.001) than in noninfected wounds and blisters according to the clinical and microbiological description of the wounds. Enzyme activities of cr

  20. Poly(dimethylsiloxane) Microchips with Two Sharpened Stretching Tips and Its Application to Protein Separation Using Dynamic Coating

    Institute of Scientific and Technical Information of China (English)

    LIU Chun-Ye; XU Xu; GAO Hong-Jun; CHEN Jie-Rong

    2007-01-01

    An integrated poly(dimethylsiloxane) (PDMS) microchip with two sharpened stretching tips for convenient sample injecting, running buffer refreshing and channel cleaning has been presented. The sample was directly introduced into the separation channel through the stretching inlet tip without complicated power switching supplies and injection cross channel. The operation of running buffer refreshing or channel cleaning was simplified by vacuuming one end of the tip and placing the other tip into the solution vial. Therefore, this fabrication method can be easily applied to most analytical laboratories economically without soft lithography and plasma bonding equipments.The attractive performance of the novel PDMS microchips has been demonstrated by using laser-induced fluorescence detection for separation of proteins. The addition of 0.04% Brij 35 in 0.04 mol/L phosphate buffer (pH 7.0)can reduce the adhesion of proteins in multienzyme tablet and make separation more easily. The electroosmotic flow (EOF) exhibits pH-independence in the range of 3-11 in dynamic modified microchannel.

  1. Simultaneous counting of two subsets of leukocytes using fluorescent silica nanoparticles in a sheathless microchip flow cytometer.

    Science.gov (United States)

    Yun, Hoyoung; Bang, Hyunwoo; Min, Junggi; Chung, Chanil; Chang, Jun Keun; Han, Dong-Chul

    2010-12-07

    A portable flow cytometer has been recognized as an important tool for many clinical applications such as HIV/AIDS screening in developing countries and regions with limited medical facilities and resources. Conventional flow cytometers typically require multiple detectors for simultaneous identification of multiple subsets of immune cell. To minimize the number of detectors toward portable flow cytometry or to analyze multi-parametric cellular information with minimum number of detectors in conventional flow cytometers, we propose a versatile multiplexed cell-counting method using functional silica nanoparticles (SiNPs). FITC-doped SiNPs, which are 100 times brighter than the FITC molecules itself, were used as new intensity-based fluorescent dye complexes to simultaneously measure two subsets of leukocytes using a single detector. CD45(+)CD4(+) cells tagged with these FITC-doped SiNPs were 50 times brighter than CD45(+)CD4(-) cells tagged only with FITC. To make the overall system compact, a disposable microchip flow cytometer that does not require sheath flow was developed. Combining these dye-doped SiNPs based detection schemes and the sheathless microchip flow cytometer scheme, we successfully identified and counted two subsets of leukocytes simultaneously (R(2) = 0.876). These approaches can be the building blocks for a truly portable and disposable flow cytometer for various clinical cytometry applications.

  2. Simple, rapid and, cost-effective fabrication of PDMS electrophoresis microchips using poly(vinyl acetate) as photoresist master.

    Science.gov (United States)

    Lobo-Júnior, Eulício O; Gabriel, Ellen F M; Dos Santos, Rodrigo A; de Souza, Fabrício R; Lopes, Wanderson D; Lima, Renato S; Gobbi, Angelo L; Coltro, Wendell K T

    2017-01-01

    This study describes a simple, rapid, and cost-effective fabrication of PDMS electrophoresis microchips using poly(vinyl acetate) (PVAc) emulsion as photoresist master. High-relief microfluidic structures were defined on poly(vinyl acetate) previously deposited on printed circuit boards surfaces without cleanroom facilities and sophisticated instrumentation. After a UV exposure, channels with heights ranging from 30 to 140 μm were obtained by controlling the emulsion mass deposited on the master surface. The developing stage was performed using water rather than the organic solvents that are applied for conventional masks. The surface morphology was characterized by optical imaging, profilometry, and SEM. Based on the achieved results, the proposed method offers suitable reproducibility for the prototyping of electrophoresis microchips in PDMS. The feasibility of the resulting PDMS electrophoresis chips was successfully demonstrated with the separation of major inorganic cations within 100 s using a contactless conductivity detection system. The separation efficiencies ranged from ca. 67 900 to 125 600 plates/m. Due to the satisfactory performance and simplified instrumentation, we believe this fabrication protocol presents potential to be implemented in any chemical, biochemical, or biological laboratory.

  3. Analysis of selective androgen receptor modulators by gas chromatography-microchip atmospheric pressure photoionization-mass spectrometry.

    Science.gov (United States)

    Luosujärvi, Laura; Haapala, Markus; Thevis, Mario; Saarela, Ville; Franssila, Sami; Ketola, Raimo A; Kostiainen, Risto; Kotiaho, Tapio

    2010-02-01

    A gas chromatography-microchip atmospheric pressure photoionization-mass spectrometric (GC-microAPPI-MS) method was developed and used for the analysis of three 2-quinolinone-derived selective androgen receptor modulators (SARMs). SARMs were analyzed from spiked urine samples, which were hydrolyzed and derivatized with N-methyl-N-(trimethylsilyl)trifluoroacetamide before analysis. Trimethylsilyl derivatives of SARMs formed both radical cations (M(+*)) and protonated molecules ([M + H](+)) in photoionization. Better signal-to-noise ratios (S/N) were obtained in MS/MS analysis using the M(+*) ions as precursor ions than using the [M + H](+) ions, and therefore the M(+*) ions were selected for the precursor ions in selected reaction monitoring (SRM) analysis. Limits of detection (LODs) with the method ranged from 0.01 to 1 ng/mL, which correspond to instrumental LODs of 0.2-20 pg. Limits of quantitation ranged from 0.03 to 3 ng/mL. The mass spectrometric response to the analytes was linear (R > or = 0.995) from the LOQ concentration level up to 100 ng/mL concentration, and intra-day repeatabilities were 5%-9%. In addition to the GC-microAPPI-MS study, the proof-of-principle of gas chromatography-microchip atmospheric pressure chemical ionization-Orbitrap MS (GC-microAPCI-Orbitrap MS) was demonstrated.

  4. Immunological cross-reactivity between four distant parvalbumins-Impact on allergen detection and diagnostics.

    Science.gov (United States)

    Sharp, Michael F; Stephen, Juan N; Kraft, Lukas; Weiss, Thomas; Kamath, Sandip D; Lopata, Andreas L

    2015-02-01

    Fish are the largest and most diverse group of vertebrates. Fish are also a part of the eight food groups that cause the majority of IgE mediated food reactions. Detection tools for fish allergens are however limited due to the great diversity of fish species, despite fish allergy and its major allergen parvalbumin being well documented. The most commonly studied fish are frequently consumed in North America and Europe. However, much less is known about fish allergens in the Australasian region although fish is widely consumed in this region. A comprehensive phylogenetic analysis was performed of known parvalbumin amino acid sequences to determine possible candidate antigens for new cross-reactive antibodies to be used to detect most fish parvalbumins. Polyclonal rabbit antibodies were raised against parvalbumins from frequently consumed barramundi (Lates calcarifer), basa (Pangasius bocourti), pilchard (Sardinops sagax) and Atlantic salmon (Salmo salar). These were evaluated for cross-reactivity against a panel of 45 fish extracts (raw, heated and canned fish). Anti-barramundi parvalbumin proved to be the most cross-reactive antibody, detecting 87.5% of the 40 species analyzed, followed by anti-pilchard and anti-basa antibody. In contrast the anti-salmon antibody was very specific and only reacted to salmonidae and a few other fish. All analyzed fish species, except mahi mahi, swordfish, yellowfin tuna and all 5 canned fish had parvalbumin detected in raw extracts. However antibody reactivity to many fish was heat liable or susceptible to denaturation, demonstrating that some parvalbumins have most likely conformational epitopes, which lose antibody reactivity after heat treatment. We have demonstrated the generation of highly cross-reactive anti-parvalbumin antibodies that could be used for the detection of allergenic fish parvalbumin in contaminated food products. This cross-reactivity study thus shows processing of fish, especially canning, can have on impact

  5. Diagnostic use of computational retrotransposon detection: Successful definition of pathogenetic mechanism in a ciliopathy phenotype.

    Science.gov (United States)

    Takenouchi, Toshiki; Kuchikata, Tomu; Yoshihashi, Hiroshi; Fujiwara, Mineko; Uehara, Tomoko; Miyama, Sahoko; Yamada, Shiro; Kosaki, Kenjiro

    2017-05-01

    Among more than 5,000 human monogenic disorders with known causative genes, transposable element insertion of a Long Interspersed Nuclear Element 1 (LINE1, L1) is known as the mechanistic basis in only 13 genetic conditions. Meckel-Gruber syndrome is a rare ciliopathy characterized by occipital encephalocele and cystic kidney disease. Here, we document a boy with occipital encephalocele, post-axial polydactyly, and multicystic renal disease. A medical exome analysis detected a heterozygous frameshift mutation, c.4582_4583delCG p.(Arg1528Serfs*17) in CC2D2A in the maternally derived allele. The further use of a dedicated bioinformatics algorithm for detecting retrotransposon insertions led to the detection of an L1 insertion affecting exon 7 in the paternally derived allele. The complete sequencing and sequence homology analysis of the inserted L1 element showed that the L1 element was classified as L1HS (L1 human specific) and that the element had intact open reading frames in the two L1-encoded proteins. This observation ranks Meckel-Gruber syndrome as only the 14th disorder to be caused by an L1 insertion among more than 5,000 known human genetic disorders. Although a transposable element detection algorithm is not included in the current best-practice next-generation sequencing analysis, the present observation illustrates the utility of such an algorithm, which would require modest computational time and resources. Whether the seemingly infrequent recognition of L1 insertion in the pathogenesis of human genetic diseases might simply reflect a lack of appropriate detection methods remains to be seen. © 2017 Wiley Periodicals, Inc.

  6. Reduction of the impedance of a contactless conductivity detector for microchip capillary electrophoresis: compensation of the electrode impedance by addition of a series inductance from a piezoelectric quartz crystal.

    Science.gov (United States)

    Kang, Qi; Shen, Dazhong; Li, Qingling; Hu, Qiang; Dong, Jianfeng; Du, Junguo; Tang, Bo

    2008-10-15

    A low-impedance capacitively coupled contactless conductivity detector (LIC (4)D) for microchip capillary electrophoresis was reported. The LIC (4)D was the series combination of a piezoelectric quartz crystal (PQC) resonator with a capacitively coupled contactless conductivity detector (C (4)D) outside on the microchip lid. The electrode impedance in the LIC (4)D was reduced because the capacitive impedance from the wall capacitance was compensated by the inductive impedance from the PQC. The operation frequency of the LIC (4)D was set at the resonant frequency of the series combination of a PQC with a C (4)D, wherein a minimum in the total impedance was obtained. It was shown that the sensitivity of LIC (4)D was much higher than that of C (4)D itself, especially in the microchip with a thick lid. Under the experimental conditions, the signal-to-noise ratios of the LIC (4)D were improved by approximately 20-50 times over those of the C (4)D. Reproducible separations of a mixture of inorganic cations (K (+), Na (+), Li (+)) were demonstrated. After a digital filter treatment by the fast Fourier transform algorithm, the detection limits were 0.38, 0.49, and 1.6 microM for K (+) in the LI C (4)D with the microchip lid thickness of 0.20, 0.40, and 1.0 mm, respectively.

  7. Diagnostic accuracy of APRI, FIB-4 and Forns for the detection of liver cirrhosis in HIV/HCV-coinfected patients.

    Science.gov (United States)

    Merli, Marco; Castagna, Antonella; Salpietro, Stefania; Gianotti, Nicola; Messina, Emanuela; Poli, Andrea; Morsica, Giulia; Bagaglio, Sabrina; Cernuschi, Massimo; Bigoloni, Alba; Uberti-Foppa, Caterina; Lazzarin, Adriano; Hasson, Hamid

    2016-04-01

    Non-invasive assessment of liver fibrosis represents an appealing method to monitor liver disease in HCV-infected patients. Currently, transient elastography (TE) is the most accurate non-invasive tool to measure liver stiffness (LS), with the diagnostic accuracy increasing together with the stage of fibrosis (Friedrich Rust et al., 2008; Degos et al., 2010). Stiffness measurement is widely used in the assessment of fibrosis in patients with chronic hepatitis C given its good reproducibility (Fraquelli et al., 2007) and its association with the risk of liver-related complications and death in HIV/HCV-coinfected patients (Fernandez-Montero et al., 2013) and also in patients with compensated HCV-related liver cirrhosis (with or without concomitant HIV-coinfection) (Pérez-Latorre et al., 2014). In the last decade, direct and indirect biomarkers for predicting liver fibrosis have also been developed. Direct fibrosis biomarkers (e.g. FibroTest, FibroMeter) are calculated using serum molecules produced in the presence of liver fibrosis and released in the circulatory system while indirect biomarkers (e.g. APRI, FIB-4, Forns) result from the combination of routine blood tests. Even though indirect biomarkers had a lower diagnostic performance than direct biomarkers and especially TE (Sánchez-Conde et al., 2010; Degos et al., 2010; Castéra et al., 2014), the absence of additional costs and the ready availability make indirect biomarkers a quick and easy non-invasive method to periodically assess liver fibrosis. Since the early detection of liver cirrhosis has a significant impact on both clinical management and treatment decision regarding chronic hepatitis C, we evaluated the threshold and the diagnostic accuracy of APRI, FIB-4 and Forns for the diagnosis of liver cirrhosis in HIV/HCV-coinfected patients.

  8. Diagnostic performance of computed tomography for detection of concomitant coronary disease in hypertrophic cardiomyopathy

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Lei; Ma, Xiaohai; Zhang, Chen; Wang, Zhanhong; Fan, Zhanming [Capital Medical University, Department of Radiology, Beijing Anzhen Hospital, Beijing (China); Ge, Hailong [Capital Medical University, Department of Cardiology, Beijing Anzhen Hospital, Beijing (China); Teraoka, Kunihiko [Tokyo Medical University, Department of Cardiology, Tokyo (Japan)

    2014-10-31

    To evaluate the diagnostic performance of computed tomography (CT) in patients with hypertrophic cardiomyopathy (HCM) and suspected coexistent coronary artery diseases (CADs). Sixty patients were enrolled in this study. Cardiac CT examination included CT coronary angiography (CTCA) and delayed enhancement CT. CT performance in evaluation of the coronary artery was assessed and compared with that of catheter-based coronary angiography (CA). The left ventricle (LV) wall thickness, functional indices and myocardial delayed enhancement (MDE) were measured via cardiac magnetic resonance (CMR) and CT images. Compared with catheter-based CA, CTCA produced a 100 % (24/24) sensitivity, a 94.4 % (34/36) specificity, a 92.3 % (24/26) positive predictive value and a 100 % (34/34) negative predictive value. CT-measured LV wall thickness and functional indices were correlated with those measured via CMR (P < 0.01), though the CT-measured values were smaller than the CMR-measured values. Bland-Altman analysis showed the volume of the focal MDE determined via CT was slightly smaller than that determined using CMR (mean difference: 0.3 cm{sup 3}). For patients with HCM and suspected coexistent CAD, this comprehensive cardiac CT protocol can be helpful in ruling out coronary stenosis and can provide information regarding morphology, function and tissue characterization of the LV myocardium. (orig.)

  9. Utility of Combination of Diagnostic Tests in Early Detection of Prostate Tumors in West Algerian Hospital

    Directory of Open Access Journals (Sweden)

    Abdelkrim Berroukche

    2014-09-01

    Full Text Available In West of Algeria, incidence of prostate cancer (PCa is growing and epidemiological data on benign prostatic hyperplasia (BPH are contradictory. The prostate specific antigen (PSA joins other tests, as digital rectal examination (DRE and ultrasound, for better management of patient. This study aimed to assess the association of previously tests with PSA-assay in diagnosis of PCa and BPH, in West Algeria. A retrospective study was performed on two groups of 234 BPH and 56 PCa diagnosed between 2010–2012 at the urology department of the hospital in Saida. Patients underwent various diagnostic tests; DRE, ultrasound, PSA and pathological examination. BPH is more common than PCa and the most dominant age group was from 70 to 79 years. DRE was positive in 67 % of BPH and 63.3 % PCa. Ultrasound examination revealed that the prostatic structure was heterogeneous in 25.5 % of BPH and 92 % PCa. Total PSA (TPSA was higher than the cut-off value of 4 ng / ml in 37% of BPH and 75 % PCa. Histological forms of Prostatic adenomyofibroma and prostatic adenocarcinoma were the most represented. This study shows that the combination of different tests is more efficient than using a test alone for a definitive diagnosis of PCa or BPH.

  10. Novel diagnostics of metabolic dysfunction detected in breath and plasma by selective isotope assisted labeling (SIAL)

    Science.gov (United States)

    Haviland, Julia A.; Tonelli, Marco; Haughey, Dermot T.; Porter, Warren P.; Assadi-Porter, Fariba M.

    2012-01-01

    OBJECTIVE Metabolomics is the study of a unique fingerprint of small molecules present in biological systems under healthy and disease conditions. One of the major challenges in metabolomics is validation of fingerprint molecules to identify specifically perturbed pathways in metabolic aberrations. This step is crucial to the understanding of budding metabolic pathologies and the ability to identify early indicators of common diseases such as obesity, diabetes mellitus type II, metabolic syndrome, polycystic ovary syndrome, and cancer. We present a novel approach to diagnosing aberrations in glucose utilization including metabolic pathway switching in a disease state. METHODS We used a well-defined prenatally exposed glucocorticoid mouse model that results in adult females with metabolic dysfunction. We applied the complementary technologies of nuclear magnetic resonance spectroscopy, and cavity ringdown spectroscopy to analyze serial plasma samples and real-time breath measurements following selective 13C-isotope assisted labeling (SIAL). These platforms allowed us to trace metabolic markers in whole animals and identify key metabolic pathway switching in prenatally glucocorticoid-treated animals. RESULTS Total glucose flux is significantly proportionally increased through the major oxidative pathways of glycolysis and the pentose phosphate pathway in the prenatally glucocorticoid-treated animals relative to the control animals. CONCLUSION This novel diagnostics approach is fast, non-invasive and sensitive for determining specific pathway utilization, and provides a direct translational application in the healthcare field. PMID:22304834

  11. Evaluation of a pan-serotype point-of-care rapid diagnostic assay for accurate detection of acute dengue infection.

    Science.gov (United States)

    Vivek, Rosario; Ahamed, Syed Fazil; Kotabagi, Shalini; Chandele, Anmol; Khanna, Ira; Khanna, Navin; Nayak, Kaustuv; Dias, Mary; Kaja, Murali-Krishna; Shet, Anita

    2017-03-01

    The catastrophic rise in dengue infections in India and globally has created a need for an accurate, validated low-cost rapid diagnostic test (RDT) for dengue. We prospectively evaluated the diagnostic performance of NS1/IgM RDT (dengue day 1) using 211 samples from a pediatric dengue cohort representing all 4 serotypes in southern India. The dengue-positive panel consisted of 179 dengue real-time polymerase chain reaction (RT-PCR) positive samples from symptomatic children. The dengue-negative panel consisted of 32 samples from dengue-negative febrile children and asymptomatic individuals that were negative for dengue RT-PCR/NS1 enzyme-linked immunosorbent assay/IgM/IgG. NS1/IgM RDT sensitivity was 89.4% and specificity was 93.8%. The NS1/IgM RDT showed high sensitivity throughout the acute phase of illness, in primary and secondary infections, in different severity groups, and detected all 4 dengue serotypes, including coinfections. This NS1/IgM RDT is a useful point-of-care assay for rapid and reliable diagnosis of acute dengue and an excellent surveillance tool in our battle against dengue. Copyright © 2016. Published by Elsevier Inc.

  12. Diagnostic utility of a direct immunofluorescence test to detect feline coronavirus antigen in macrophages in effusive feline infectious peritonitis.

    Science.gov (United States)

    Litster, A L; Pogranichniy, R; Lin, T-L

    2013-11-01

    The antemortem diagnosis of feline infectious peritonitis (FIP) remains challenging in clinical practice, since current testing methods have suboptimal diagnostic accuracy. Immunohistochemical testing of biopsy specimens and postmortem examination are the standard diagnostic methods, although direct immunofluorescence (DIF) testing to detect feline coronavirus in macrophages in effusion specimens has been reported to have 100% specificity and has been recommended as an antemortem confirmatory test. The aim of this study was to compare the results of DIF testing in antemortem feline effusions with postmortem results using field samples. Effusion specimens were collected antemortem from 17 cats and tested by DIF, followed by postmortem examination. Histopathological examination of specimens collected at postmortem confirmed FIP in 10/17 cases and ruled out FIP out in 7/17 cases. Antemortem DIF testing was positive in all 10 cases confirmed as FIP at postmortem examination. In the seven cats where FIP was ruled out at postmortem examination, DIF was negative in five cases and positive in the remaining two cases. The calculated sensitivity of DIF testing was 100% and the specificity was 71.4%. Duplicate effusion specimens from eight cats that were initially DIF positive were stored refrigerated (4 °C) or at room temperature (22-25 °C) and subjected to serial DIF testing to determine the duration of positive results. DIF-positive specimens stored at both temperatures retained their positive status for at least 2 days.

  13. Detection of collagen by second harmonic microscopy as a diagnostic tool for liver fibrosis

    Science.gov (United States)

    Banavar, Maruth; Kable, Eleanor P. W.; Braet, Filip; Wang, X. M.; Gorrell, M. D.; Cox, Guy

    2006-02-01

    Liver fibrosis has many causes, including hepatitis C, alcohol abuse, and non-alcoholic steatohepatitis. It is characterized by abnormal deposition of extracellular matrix proteins, mainly collagen. The deposition of these proteins results in impaired liver function caused by distortion of the hepatic architecture by fibrous scar tissue. The unique triple helix structure of collagen and high level of crystallinity make it very efficient for generating second harmonic signals. In this study we have set out to see if second harmonic imaging of collagen can be used as a non-biased quantitative tool for classification of fibrosis levels in liver biopsies and if it can detect early fibrosis formation not detected by current methods.

  14. Diagnostic efficacy of Ziehl-Neelsen method against fluorescent microscopy in detection of acid fast bacilli

    Institute of Scientific and Technical Information of China (English)

    Soham Gupta; Vishnu Prasad Shenoy; Indira Bairy; MuralidharanS

    2010-01-01

    Objective:To investigate the application of Ziehl-Neelsen (Z-N) and fluorescent microscopy in detection of acid fast bacilli (AFB).Methods: Duplicate smears were prepared from 260 sputum samples and stained with Z-N and fluorescent staining (FS) methods. The efficiency of both methods in primary diagnosis of tuberculosis were evaluated.Results:The smears were positive for AFB in 15 (5.77%) samples by Z-N staining method and in 16 (6.15%) samples by FS method. The sensitivity and specificity of Z-N staining method against FS method were 93.75% and 100% respectively.Conclusions: Though lesser cost-effective than Z-N, FS method is a more sensitive and better case finding tool in detection of AFB.

  15. Detection of Pneumocystis jirovecii in oral wash from immunosuppressed patients as a diagnostic tool

    DEFF Research Database (Denmark)

    Hviid, Cecilie Juul; Lund, Marianne; Sørensen, Allan Klitgaard

    2017-01-01

    BACKGROUND: Diagnosis of Pneumocystis jirovecii (PJ) pneumonia ordinarily requires invasive procedures that could be avoided by PCR methodologies, if these could be designed with adequate cut-off values for confounding background carriage. METHODS: We designed a novel quantitative real-time PCR......), rheumatologic patients (n = 102), patients with inflammatory bowel diseases (n = 98), and healthy blood donors (controls, n = 50). The fungal burden in patients with PJ pneumonia (PCP, n = 7) was also investigated. RESULTS: Danish HIV-infected patients (with viremia/low CD4) and recent transplant recipients...... were at most risk of being carriers (prevalence of 23% and 16.7% respectively), whereas PJ was rarely detected among rheumatologic patients, patients with inflammatory bowel diseases, and untreated West African HIV patients. PJ was not detected among healthy controls. The fungal burden in patients...

  16. Spectroscopic techniques as a diagnostic tool for early detection of osteoporosis

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Kanika; Lee, Kwang Sung; Lee, Dong Geun; Kim, Yong Ki; Kim, Kyung Chun [Pusan National University, Busan (Korea, Republic of)

    2010-08-15

    Osteoporosis (OP) a kind of bone disease, is very serious in particular for old persons, and may lead them to immobility and death. Early detection of the diseases is the first consideration for the patients to have more options to live a healthy life. The biomarkers or bonemarkers provide a promising challenge in clinical proteomics for early disease detection. In this paper, optical techniques such as Fourier Transform Infrared Spectroscopy (FTIR) and UV/Visible spectroscopy are employed to find the bone markers and emphasis has been given on noninvasive modalities for early detection of osteoporosis. Blood plasma samples procured from two groups, patients and healthy persons were tested. Both of the optical techniques revealed obvious differences in the spectra: between two groups, for example, increase in intensity for OP persons. New peaks were found at 1646, 1540, 1456 and 1077 cm-1 in FTIR spectra. Except 1588 cm-1, we showed decrease in spectral intensity of OP persons. In UV/Visible spectroscopy results, new peaks appeared in the OP patients spectra at the wavelength of 279 nm and 414 nm. These differences in the spectra of the two types samples, allow rapid and cost-effective discrimination of the potential patients with the optical techniques which were verified by the bone densitometer in the hospitals. The new and novel technique is quick, reliable and effective

  17. Hybrid Model-Based and Data-Driven Fault Detection and Diagnostics for Commercial Buildings: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Frank, Stephen; Heaney, Michael; Jin, Xin; Robertson, Joseph; Cheung, Howard; Elmore, Ryan; Henze, Gregor

    2016-08-01

    Commercial buildings often experience faults that produce undesirable behavior in building systems. Building faults waste energy, decrease occupants' comfort, and increase operating costs. Automated fault detection and diagnosis (FDD) tools for buildings help building owners discover and identify the root causes of faults in building systems, equipment, and controls. Proper implementation of FDD has the potential to simultaneously improve comfort, reduce energy use, and narrow the gap between actual and optimal building performance. However, conventional rule-based FDD requires expensive instrumentation and valuable engineering labor, which limit deployment opportunities. This paper presents a hybrid, automated FDD approach that combines building energy models and statistical learning tools to detect and diagnose faults noninvasively, using minimal sensors, with little customization. We compare and contrast the performance of several hybrid FDD algorithms for a small security building. Our results indicate that the algorithms can detect and diagnose several common faults, but more work is required to reduce false positive rates and improve diagnosis accuracy.

  18. Hybrid Model-Based and Data-Driven Fault Detection and Diagnostics for Commercial Buildings

    Energy Technology Data Exchange (ETDEWEB)

    Frank, Stephen; Heaney, Michael; Jin, Xin; Robertson, Joseph; Cheung, Howard; Elmore, Ryan; Henze, Gregor

    2016-08-26

    Commercial buildings often experience faults that produce undesirable behavior in building systems. Building faults waste energy, decrease occupants' comfort, and increase operating costs. Automated fault detection and diagnosis (FDD) tools for buildings help building owners discover and identify the root causes of faults in building systems, equipment, and controls. Proper implementation of FDD has the potential to simultaneously improve comfort, reduce energy use, and narrow the gap between actual and optimal building performance. However, conventional rule-based FDD requires expensive instrumentation and valuable engineering labor, which limit deployment opportunities. This paper presents a hybrid, automated FDD approach that combines building energy models and statistical learning tools to detect and diagnose faults noninvasively, using minimal sensors, with little customization. We compare and contrast the performance of several hybrid FDD algorithms for a small security building. Our results indicate that the algorithms can detect and diagnose several common faults, but more work is required to reduce false positive rates and improve diagnosis accuracy.

  19. Astrovirus Diagnostics

    Science.gov (United States)

    Pérot, Philippe; Lecuit, Marc; Eloit, Marc

    2017-01-01

    Various methods exist to detect an astrovirus infection. Current methods include electron microscopy (EM), cell culture, immunoassays, polymerase chain reaction (PCR) and various other molecular approaches that can be applied in the context of diagnostic or in surveillance studies. With the advent of metagenomics, novel human astrovirus (HAstV) strains have been found in immunocompromised individuals in association with central nervous system (CNS) infections. This work reviews the past and current methods for astrovirus detection and their uses in both research laboratories and for medical diagnostic purposes. PMID:28085120

  20. Diagnostic Accuracy of CBCT with Different Voxel Sizes and Intraoral Digital Radiography for Detection of Periapical Bone Lesions: An Ex-Vivo Study

    OpenAIRE

    Shirin Sakhdari; Ahmad Reza Talaeipour; Maziar Talaeipour; Maryam Pazhutan; Sanaz Heidarkhan Tehrani; Mohammad Javad Kharazifard

    2016-01-01

    Objectives: This study sought to assess the diagnostic accuracy of cone beam computed tomography (CBCT) with different voxel sizes and intraoral digital radiography with photostimulable phosphor (PSP) plate for detection of periapical (PA) bone lesions.Materials and Methods: In this ex vivo diagnostic study, one-millimeter defects were created in the alveolar sockets of 15 bone blocks, each with two posterior teeth. A no-defect control group was also included. Digital PA radiographs with PSP ...

  1. Serologic and Urine Diagnostic Tests to Detect Helicobacter pylori Infection in Functional Dyspepsia Patients

    Directory of Open Access Journals (Sweden)

    Agasjtya Wisjnu Wardhana

    2016-12-01

    Full Text Available Background: Dyspepsia is a collection of symptoms in the forms of discomfort, pain, nausea, vomiting, bloating, and early satiety in the stomach. This condition can be caused by various problems; one of them is Helicobacter pylori infection. Dyspepsia without organic problem is known as functional dyspepsia. H. pylori examination is recommended in functional dyspepsia patients. Method: In this study, we performed a diagnostic test study in dyspepsia patients in Community Health Centre of Koja District, North Jakarta, from February to April 2015. Samples were obtained through consecutive sampling method; 74 patients were included. The data was gathered by distributing questionnaires to patients, performing urea breath test (UBT examination, serologic test, and urine test using rapid urine test (RAPIRUN. Results: Prevalence of H. pylori infection by using UBT examination reached up to 36.5%; meanwhile serologic and RAPIRUN tests showed positive results in 32.4% and 24.3% patients, respectively. Serologic test has sensitivity of 74% (95% CI: 55-87%, specificity 91% (95% CI: 80-97%, positive predictive value (PPV 83% (95% CI: 64-93%, and negative predictive value (NPV 86% (95% CI: 74-93%. Meanwhile, RAPIRUN has sensitivity of 63% (95% CI: 44-78%, specificity 98% (95% CI: 89-100%, PPV 94% (95% CI: 74-99%, and NPV 82% (95% CI: 70-90%. Conclusion: Sensitivity of serologic and RAPIRUN tests are still inadequate to be alternative to UBT examination. However, they have high specificity. Further studies are required with larger sample size and consideration of factors which may influence the results of both tests.

  2. Diagnostic performance of direct traction MR arthrography of the hip: detection of chondral and labral lesions with arthroscopic comparison

    Energy Technology Data Exchange (ETDEWEB)

    Schmaranzer, Florian; Klauser, Andrea; Henninger, Benjamin [Medical University Innsbruck, Department of Radiology, Innsbruck (Austria); Kogler, Michael; Schmaranzer, Ehrenfried [District Hospital St. Johann in Tyrol, Department of Radiology, St. Johann in Tyrol (Austria); Forstner, Thomas [Johannes Keppler University, Department for Applied Systems Research and Statistics, Linz (Austria); Reichkendler, Markus [District Hospital St. Johann in Tyrol, Department of Orthopedic Surgery, St. Johann in Tyrol (Austria)

    2015-06-01

    To assess diagnostic performance of traction MR arthrography of the hip in detection and grading of chondral and labral lesions with arthroscopic comparison. Seventy-five MR arthrograms obtained ± traction of 73 consecutive patients (mean age, 34.5 years; range, 14-54 years) who underwent arthroscopy were included. Traction technique included weight-adapted traction (15-23 kg), a supporting plate for the contralateral leg, and intra-articular injection of 18-27 ml (local anaesthetic and contrast agent). Patients reported on neuropraxia and on pain. Two blinded readers independently assessed femoroacetabular cartilage and labrum lesions which were correlated with arthroscopy. Interobserver agreement was calculated using κ values. Joint distraction ± traction was evaluated in consensus. No procedure had to be stopped. There were no cases of neuropraxia. Accuracy for detection of labral lesions was 92 %/93 %, 91 %/83 % for acetabular lesions, and 92 %/88 % for femoral cartilage lesions for reader 1/reader 2, respectively. Interobserver agreement was moderate (κ = 0.58) for grading of labrum lesions and substantial (κ = 0.7, κ = 0.68) for grading of acetabular and femoral cartilage lesions. Joint distraction was achieved in 72/75 and 14/75 hips with/without traction, respectively. Traction MR arthrography safely enabled accurate detection and grading of labral and chondral lesions. (orig.)

  3. ProEx C as Diagnostic Marker for Detection of Urothelial Carcinoma in Urinary Samples: A Review.

    Science.gov (United States)

    Botti, Gerardo; Malzone, Maria Gabriella; La Mantia, Elvira; Montanari, Micaela; Vanacore, Daniela; Rossetti, Sabrina; Quagliariello, Vincenzo; Cavaliere, Carla; Di Franco, Rossella; Castaldo, Luigi; Ametrano, Gianluca; Cappuccio, Francesca; Romano, Francesco Jacopo; Piscitelli, Raffaele; Pepe, Maria Filomena; D'Aniello, Carmine; Facchini, Gaetano

    2017-01-01

    The gold standard for the detection of urothelial carcinoma is represented by urethro-cystoscopy and biopsy. Both procedures are invasive and expensive and therefore cytology is often used as first approach to investigate on a possible neoplasia, being a safe and cost-effective diagnostic modality of evaluation. Because cytology alone is not highly sensitive for detection of low grade urothelial carcinoma and recurrence of the disease, several adjunct markers and urine based tests for urothelial carcinoma have been developed, which can help in the final diagnosis. In particular, ProEx C is an immunohistochemical cocktail containing antibodies direct against topoisomerase IIα (TOP2A) and minichromosome maintenance 2 (MCM2) proteins. It proved to be a valid biomarker especially in detecting squamous intraepithelial lesions in cervical liquid-based samples and in discerning these lesions from their mimickers, as well as in ovarian, endometrial, vulvar, primary and metastatic melanomas, breast, pancreatic and renal cell carcinomas. This brief review covers the effective utility of ProEx C as adjunct tool in assessing the urothelial lesions in urine cytology, also providing prognostic and therapeutic information to help in clinical decisions.

  4. Direct Determination of a Small-Molecule Drug, Valproic Acid, by an Electrically-Detected Microcantilever Biosensor for Personalized Diagnostics

    Directory of Open Access Journals (Sweden)

    Long-Sun Huang

    2015-01-01

    Full Text Available Direct, small-molecule determination of the antiepileptic drug, valproic acid, was investigated by a label-free, nanomechanical biosensor. Valproic acid has long been used as an antiepileptic medication, which is administered through therapeutic drug monitoring and has a narrow therapeutic dosage range of 50–100 μg·mL−1 in blood or serum. Unlike labeled and clinically-used measurement techniques, the label-free, electrical detection microcantilever biosensor can be miniaturized and simplified for use in portable or hand-held point-of-care platforms or personal diagnostic tools. A micromachined microcantilever sensor was packaged into the micro-channel of a fluidic system. The measurement of the antiepileptic drug, valproic acid, in phosphate-buffered saline and serum used a single free-standing, piezoresistive microcantilever biosensor in a thermally-controlled system. The measured surface stresses showed a profile over a concentration range of 50–500 μg·mL−1, which covered the clinically therapeutic range of 50–100 μg·mL−1. The estimated limit of detection (LOD was calculated to be 45 μg·mL−1, and the binding affinity between the drug and the antibody was measured at around 90 ± 21 μg·mL−1. Lastly, the results of the proposed device showed a similar profile in valproic acid drug detection with those of the clinically-used fluorescence polarization immunoassay.

  5. Evaluation of Diagnostic Sensitivity of Wet Preparation Microscopy Using KOH for Detection of Fungal Agents from Keratitis Patients

    Directory of Open Access Journals (Sweden)

    Neeta D Khokhar, Summaiya A Mulla, Latika N Shah, Geeta M Vaghela

    2013-01-01

    Full Text Available Background: Keratitis is a common ophthalmic condition mostly caused by fungi. Apart from fungal culture, wet preparation using 10% Potassium hydroxide (KOH for microscopic detection of fungal elements is a rapid and accurate method of laboratory diagnosis. Purpose: This prospective study was undertaken in order to evaluate the diagnostic sensitivity of wet preparation microscopy using KOH for detection of fungal agents from keratitis patients. Methodology: 103 samples of clinically suspected patients of keratitis attending tertiary care hospital between march 2010 and june 2011 were included. Samples like corneal swabs, corneal scrapings, corneal button, and corneo-scleral rim were collected aseptically after slit lamp examination, then transported to microbiology laboratory. Samples were processed for direct microscopy (gram stain and 10% KOH wet mount preparation and culture. Culture positive isolates were identified based on morphology and standard biochemical tests. Data entry and analysis was done statistically. Results: From 103 samples, fungal culture was positive in 12%. Different fungus isolates include Aspergillus flavus (67% and Candida spp (25% were the leading fungi followed by Curvularis (8%. Direct microscopical examination using KOH wet preparation and gram’s stain had detected fungal elements in 83% and 75% samples respectively against culture results. Conclusion: Aspergillus flavus (67%, Candida spp (25% and Curvularia spp (8% was most common cause of fungal keratitis. Wet mount with KOH can be relied upon as the single most important screening tool for rapid diagnosis of fungal corneal ulcer and treatment should be dispensed on its basis.

  6. Diagnostic Accuracy of Five Different Fecal Markers for the Detection of Precancerous and Cancerous Lesions of the Colorectum

    Directory of Open Access Journals (Sweden)

    Mariann Rutka

    2016-01-01

    Full Text Available Background. Colorectal cancer (CRC is the second deadliest malignancy worldwide. This study aimed to compare the diagnostic accuracy of different fecal markers in the detection of colorectal adenomas and cancer. Methods. Stool samples of patients referred to colonoscopy were collected for the analysis of tumor M2 pyruvate kinase (M2PK, human hemoglobin (Hb, hemoglobin/haptoglobin (Hb/Hp complex, fecal calprotectin (FC, and matrix metalloproteinase-9 (MMP-9. Results. Sensitivity and specificity of M2PK for adenomas sized > 1 cm were 60% and 67.5% and for CRC were 94.7% and 67.5%. Sensitivity and specificity of iFOBT for adenomas sized ≥ 1 cm were 80% and 72.5% and for CRC were 94.7% and 72.5%. Sensitivity and specificity of Hb/Hp complex for adenomas sized ≥ 1 cm were 80% and 52.9% and for CRC were 100% and 52.9%. Sensitivity of FC and MMP-9 for CRC was 77.8% and 72.2%. Combined use of M2PK, iFOBT, and FC resulted in a sensitivity and specificity of 95% and 47.5% for the detection of adenomas sized ≥ 1 cm. Discussion. In CRC, sensitivity of M2PK, iFOBT, and Hb/Hp complex proved to be high. Combined use of M2PK, iFOBT, and FC may be valuable in the detection of large adenomas.

  7. Laboratory diagnostic outcome applying detection criteria recommended by the Scientific and Standardization Committee of the ISTH on Lupus Anticoagulant.

    Science.gov (United States)

    Chantarangkul, Veena; Biguzzi, Eugenia; Asti, Daniela; Palmucci, Claudia; Tripodi, Armando

    2013-07-01

    This study shows the diagnostic outcome of an APTT-based and two dRVVT-based commercial confirmatory integrated tests with the application of the recommendations by the Scientific and Standardization Committee (SSC) on Lupus anticoagulant (LA)/antiphospholipid syndrome (APS) of the International Society on Thrombosis and Haemostasis (ISTH) issued in 2009 concerning the cut-off values for the screening, mixing and confirmatory tests for the detection of LA and the mandatory need to perform mixing tests of patient plasma with pooled normal plasma. The study population included 565 patients collected from a large central coagulation laboratory, for which the attending physicians requested LA detection. One-hundred-six healthy subjects (HS) and 131 selected patients on oral anticoagulant therapy (OAT) were included as negative controls. The results suggest that the performance of mixing tests is indicated for those methods with relatively poor specificity, but is less needed for those methods with high specificity. Furthermore, the SSC recommendation to use normal mid-value (i.e. the 50th percentile of distribution of results from healthy subjects) as the cut-off to interpret results of confirmatory tests, showed a modest increase in LA detection rate (sensitivity) but at the expense of specificity, particularly in methods with low specificity.

  8. Development of Rapid Diagnostic Kit for Identification of Hanwoo (Korean Native Cattle) Brand Meat by Detecting BIO-TAG.

    Science.gov (United States)

    Baek, Kyung Hoon; Park, Sung Kwon; Lee, Myung Hoon; Kim, Sung Il; Cho, Soo Hyun; Choi, Chang Bon

    2014-01-01

    This study was performed to develop a rapid immuno-assay kit, by using a specific antigen to detect Hanwoo brand meat. We selected a synthetic antigen specific to our target antibody, named BIO-TAG (Tyr-D-Ala-Phe), by utilizing a computer-based analysis and literature review. BIO-TAG tagged with adjuvant was subcutaneously injected in sheep and Hanwoo. The serum and meat juice of the immunized or non-immunized animal were then analyzed, to measure the titer of antibody by ELISA and Western blot. The amount of antibodies against the BIO-TAG increased (pbrand meat can be more precisely identified by our rapid diagnostic kit. This technology can deter possible fraud of counterfeit meat brands in the Korean domestic market with ease and rapidity; and offers a new tool that guarantees consumers high quality Hanwoo brand beef.

  9. Comparison of murex single-use diagnostic system with traditional enzyme immunoassay for detection of exposure to human immunodeficiency virus.

    Science.gov (United States)

    Martin, Christin A; Keren, David F

    2002-01-01

    Because a retrospective study detected 13 negative Western blots out of 38 single-use diagnostic system (SUDS)-positive cases over a 1-year period, we performed a prospective study to compare the performance of the SUDS test with that of enzyme immunoassay (EIA). Of 888 SUDS-tested sera, 875 (98.4%) were both SUDS and EIA negative and 5 (0.6%) were SUDS, EIA, and Western blot positive. The rate of SUDS-positive samples decreased from 3.16/month in the retrospective study to 1.33/month in the prospective study. The immunoassays had sensitivities and specificities of 100 and 99.7 (SUDS) and 100 and 99.4% (traditional EIA), respectively. In laboratories with experienced personnel, the SUDS test performs as well as the EIA as a screen for infection with the human immunodeficiency virus.

  10. A study on the applicability of implantable microchip transponders for body temperature measurements in pigs

    DEFF Research Database (Denmark)

    Lohse, Louise; Uttenthal, Åse; Enøe, Claes

    2010-01-01

    Background The applicability of an electronic monitoring system using microchip transponders for measurement of body temperatures was tested in 6-week-old conventional Danish weaners infected with classical swine fever virus (CSFV). Subcutaneous tissue temperatures obtained by the implantable...

  11. Acupuncture injection for field amplified sample stacking and glass microchip-based capillary gel electrophoresis.

    Science.gov (United States)

    Ha, Ji Won; Hahn, Jong Hoon

    2017-02-01

    Acupuncture sample injection is a simple method to deliver well-defined nanoliter-scale sample plugs in PDMS microfluidic channels. This acupuncture injection method in microchip CE has several advantages, including minimization of sample consumption, the capability of serial injections of different sample solutions into the same microchannel, and the capability of injecting sample plugs into any desired position of a microchannel. Herein, we demonstrate that the simple and cost-effective acupuncture sample injection method can be used for PDMS microchip-based field amplified sample stacking in the most simplified straight channel by applying a single potential. We achieved the increase in electropherogram signals for the case of sample stacking. Furthermore, we present that microchip CGE of ΦX174 DNA-HaeⅢ digest can be performed with the acupuncture injection method on a glass microchip while minimizing sample loss and voltage control hardware. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Effect of mixing on reaction-diffusion kinetics for protein hydrogel-based microchips.

    Science.gov (United States)

    Zubtsov, D A; Ivanov, S M; Rubina, A Yu; Dementieva, E I; Chechetkin, V R; Zasedatelev, A S

    2006-03-09

    Protein hydrogel-based microchips are being developed for high-throughput evaluation of the concentrations and activities of various proteins. To shorten the time of analysis, the reaction-diffusion kinetics on gel microchips should be accelerated. Here we present the results of the experimental and theoretical analysis of the reaction-diffusion kinetics enforced by mixing with peristaltic pump. The experiments were carried out on gel-based protein microchips with immobilized antibodies under the conditions utilized for on-chip immunoassay. The dependence of fluorescence signals at saturation and corresponding saturation times on the concentrations of immobilized antibodies and antigen in solution proved to be in good agreement with theoretical predictions. It is shown that the enhancement of transport with peristaltic pump results in more than five-fold acceleration of binding kinetics. Our results suggest useful criteria for the optimal conditions for assays on gel microchips to balance high sensitivity and rapid fluorescence saturation kinetics.

  13. Effects of improved microchannel structures on the separation characteristics of microchip capillary electrophoresis

    CERN Document Server

    Utsumi, Y; Ozaki, M; Terabe, S

    2003-01-01

    We fabricated the electrophoresis microchips using the UV polymerization technique. We employed plastic substrates that were suitable for rapid prototyping instead of glass and quartz. A thick UV negative photo resist was used to form molds and poly-dimethylsilozane (PDMS) was polymerized by a thermal curing process on the mold to obtain replica microchips. Electroosmotic flow (EOF) was measured to evaluate the surface. Rhodamine B and sulforhodamine B are successfully separated using the microchip. Characteristic differences between UV-fabricated and SR-fabricated microchips were evaluated by EOF measurement. It was observed that accurately defined microchannels fabricated by synchrotron radiation (SR) lithography show constant peak heights and FWHMs. Thus the advantage of the application of SR lithography to the mold fabrication is also demonstrated. (author)

  14. Kinetic effects on signal normalization in oligonucleotide microchips with labeled immobilized probes.

    Science.gov (United States)

    Pan'kov, S V; Chechetkin, V R; Somova, O G; Antonova, O V; Moiseeva, O V; Prokopenko, D V; Yurasov, R A; Gryadunov, D A; Chudinov, A V

    2009-10-01

    Among various factors affecting operation of oligonucleotide microchips, the variations in concentration and in homogeneous distribution of immobilized probes over the cells are one of the most important. The labeling of immobilized probes ensures the complete current monitoring on the probe distribution and is reliable and convenient. Using hydrogel-based oligonucleotide microchips, the applicability of Cy3-labeled immobilized probes for quality control and signal normalization after hybridization with Cy5-labeled target DNA was investigated. This study showed that proper signal normalization should be different in thermodynamic conditions and in transient regime with hybridization far from saturation. This kinetic effect holds for both hydrogel-based and surface oligonucleotide microchips. Besides proving basic features, the technique was assessed on a sampling batch of 50 microchips developed for identifying mutations responsible for rifampicin and isoniazid resistance of Mycobacterium tuberculosis.

  15. Optical Fiber Lasers and All Solid-State Passively Modulated Microchip Lasers

    Institute of Scientific and Technical Information of China (English)

    Junewen; Chen; Pie-Yau; Chien; Yu-Ting; Lee

    2003-01-01

    Erbium fiber lasers of continuous mode outputs and of pulsed picosecond and sub-picosecond pulses train are reported. Compact all solid state passively modulated microchip lasers are also developed to the same degree.

  16. Clinical diagnostic value of Molybdenum Target X-ray combined with four Serum tumor markers in the detection of Mastocarcinoma

    Institute of Scientific and Technical Information of China (English)

    Ding-Hua Xu; Chun-Xian Cai; Guang-Xue Huang

    2015-01-01

    Objective: To explore the clinical diagnostic value of Molybdenum Target X-ray combined with serum CA15-3, CA125, CEA and CYFRA21-1 tumor markers in the detection of Mastocarcinoma. Method:90 cases confirmed by surgery pathology diagnosis according to the diagnosis of breast nodules disease patients were divided into benign tumor group (n=32) and breast cancer group (n=58), all patients were with preoperative line of molybdenum target X-ray radiography examination; Another group chose healthy women of our hospital for check-up in 56 cases as the control group, electrochemiluminescence immunoassay detection of three groups of participants were four kinds of serum tumor markers level, evaluation of single and combined testing the sensitivity of the diagnosis of breast cancer, specific degree, positive predictive value and negative predictive value. Results: CA15-3, CA125, CEA and CYFRA21-1 in breast cancer group were significantly higher than that of the control group and benign tumor group, the difference was statistically significant (all P0.05); Molybdenum target X-ray slice of the sensitivity of the diagnosis of breast cancer was higher, but the specific degree was low, and lower the sensitivity of the serum tumor markers in the diagnosis of breast cancer, the specific degree was higher, the molybdenum target X-ray slice joint the sensitivity of the four tumor markers in the diagnosis of breast cancer was 89.66%, 78.13%. Conclusions: The combined Mastocarcinoma detection of Molybdenum Target X-ray with serum CA15-3, CA125, CEA and CYFRA21-1 tumor markers can improve the detective rate and plays an important role in Mastocarcinoma early detection.

  17. A comparative study of the diagnostic accuracy on Waters view with CT scan in detecting midface fractures

    Directory of Open Access Journals (Sweden)

    Panjnoush M.

    2006-08-01

    Full Text Available Background and Aim: In recent years, CT scan has become available as an alternative to conventional radiography. To date, the utility of Waters view in detecting midface fractures has been rarely evaluated. The aim of this study was to compare the diagnostic accuracy and reliability of Waters radiography with CT scan in detecting midface fractures. Materials and Methods: In this tests evaluation study, waters view and CT scan were performed for 42 patients with midface fracture admitted to maxillofacial surgery department of Shariati hospital. All images were observed and interpreted by an oral and maxillofacial radiologist and an oral and maxillofacial surgeon. Sensitivity, specificity and reliability for Waters view in detecting midface fractures were assessed by Cohen’s kappa test. Results: Sensitivity and specificity for Waters view in detection of midface fratures by the radiologist were 31.79% and 95.35% and by the surgeon were 29.59% and 93.75% respectively. The highest reliability in CT scan and Waters view (in nasal fractures by the radiologist was 66.67% and was 58.33% by the surgeon in buttress of zygoma. The highest agreement rate between the radiologist and the surgeon for CT scan was in zygomatic arch (78.95% and for Waters view was in nasal fracture (62.5%. Conclusion: Based on the results of this study, the specificity of Waters view is sufficient to diagnose fractures of lateral orbital wall, infraorbital rim, orbital floor, zygomatic arch, frontozygomatic suture, lateral wall of maxillary sinus and Lefort II fracture. The specificity is not sufficient to diagnose fractures of medial orbital wall and anterior, posterior and medial wall of maxillary sinus. Detection of these midface fractures needs other conventional radiographies or CT scan.

  18. Effect of Time Lapse on the Diagnostic Accuracy of Cone Beam Computed Tomography for Detection of Vertical Root Fractures.

    Science.gov (United States)

    Eskandarloo, Amir; Asl, Amin Mahdavi; Jalalzadeh, Mohsen; Tayari, Maryam; Hosseinipanah, Mohammad; Fardmal, Javad; Shokri, Abbas

    2016-01-01

    Accurate and early diagnosis of vertical root fractures (VRFs) is imperative to prevent extensive bone loss and unnecessary endodontic and prosthodontic treatments. The aim of this study was to assess the effect of time lapse on the diagnostic accuracy of cone beam computed tomography (CBCT) for VRFs in endodontically treated dog's teeth. Forty-eight incisors and premolars of three adult male dogs underwent root canal therapy. The teeth were assigned to two groups: VRFs were artificially induced in the first group (n=24) while the teeth in the second group remained intact (n=24). The CBCT scans were obtained by NewTom 3G unit immediately after inducing VRFs and after one, two, three, four, eight, 12 and 16 weeks. Three oral and maxillofacial radiologists blinded to the date of radiographs assessed the presence/absence of VRFs on CBCT scans. The sensitivity, specificity and accuracy values were calculated and data were analyzed using SPSS v.16 software and ANOVA. The total accuracy of detection of VRFs immediately after surgery, one, two, three, four, eight, 12 and 16 weeks was 67.3%, 68.7%, 66.6%, 64.6%, 64.5%, 69.4%, 68.7%, 68% respectively. The effect of time lapse on detection of VRFs was not significant (p>0.05). Overall sensitivity, specificity and accuracy of CBCT for detection of VRFs were 74.3%, 62.2%, 67.2% respectively. Cone beam computed tomography is a valuable tool for detection of VRFs. Time lapse (four months) had no effect on detection of VRFs on CBCT scans.

  19. Effect of time lapse on the diagnostic accuracy of cone beam computed tomography for detection of vertical root fractures

    Energy Technology Data Exchange (ETDEWEB)

    Eskandarloo, Amir; Shokri, Abbas, E-mail: Dr.a.shokri@gmail.com [Dental Research Center, Department of Oral and Maxillofacial Radiology, Hamadan University of Medical Sciences, Hamadan (Iran, Islamic Republic of); Asl, Amin Mahdavi [Department of Oral and Maxillofacial Radiology, Golestan University of Medical Sciences, Gorgan (Iran, Islamic Republic of); Jalalzadeh, Mohsen [Department of Endodontics, Hamadan University of Medical Sciences, Hamadan (Iran, Islamic Republic of); Tayari, Maryam [Department of Pedodontics, Golestan University of Medical Sciences, Gorgan (Iran, Islamic Republic of); Hosseinipanah, Mohammad [Department of Anatomy, School of Medicine, Hamadan University of Medical Sciences, Hamadan (Iran, Islamic Republic of); Fardmal, Javad [Research Center for Health Sciences and Department of Epidemiology and Biostatistics, School of Public Health, Hamadan University of Medical Sciences, Hamadan (Iran, Islamic Republic of)

    2016-01-15

    Accurate and early diagnosis of vertical root fractures (VRFs) is imperative to prevent extensive bone loss and unnecessary endodontic and prosthodontic treatments. The aim of this study was to assess the effect of time lapse on the diagnostic accuracy of cone beam computed tomography (CBCT) for VRFs in endodontically treated dog’s teeth. Forty eight incisors and premolars of three adult male dogs underwent root canal therapy. The teeth were assigned to two groups: VRFs were artificially induced in the first group (n=24) while the teeth in the second group remained intact (n=24). The CBCT scans were obtained by NewTom 3G unit immediately after inducing VRFs and after one, two, three, four, eight, 12 and 16 weeks. Three oral and maxillofacial radiologists blinded to the date of radiographs assessed the presence/absence of VRFs on CBCT scans. The sensitivity, specificity and accuracy values were calculated and data were analyzed using SPSS v.16 software and ANOVA. The total accuracy of detection of VRFs immediately after surgery, one, two, three, four, eight, 12 and 16 weeks was 67.3%, 68.7%, 66.6%, 64.6%, 64.5%, 69.4%, 68.7%, 68% respectively. The effect of time lapse on detection of VRFs was not significant (p>0.05). Overall sensitivity, specificity and accuracy of CBCT for detection of VRFs were 74.3%, 62.2%, 67.2% respectively. Cone beam computed tomography is a valuable tool for detection of VRFs. Time lapse (four months) had no effect on detection of VRFs on CBCT scans. (author)

  20. Diagnostic Accuracy of Methylated SEPT9 for Blood-based Colorectal Cancer Detection: A Systematic Review and Meta-Analysis

    Science.gov (United States)

    Nian, Jiayun; Sun, Xu; Ming, SuYang; Yan, Chen; Ma, Yunfei; Feng, Ying; Yang, Lin; Yu, Mingwei; Zhang, Ganlin; Wang, Xiaomin

    2017-01-01

    Objectives: More convenient and effective blood-based methods are believed to increase colorectal cancer (CRC) detection adoption. The effectiveness of methylated SPET9 for CRC detection has been reviewed in the newly published recommendation statement by US Preventive Services Task Force (USPSTF), while detailed instructions were not provided, which may be a result of insufficient evidence. Therefore, more evidence is needed to assist practitioners to thoroughly understand the utilization of this special maker. Methods: Based on the standard method, a systematic review and meta-analysis was performed. Quadas-2 was used to assess the methodological quality of studies. Relevant studies were searched and screened from PubMed, Embase and other literature databases up to June 1, 2016. Pooled sensitivity, specificity and diagnostic odds ratio were summarized by bivariate mixed effect model and area under the curve (AUC) was estimated by hierarchical summary receiver operator characteristic curve. Results: 25 studies were included for analysis. The pooled sensitivity, specificity and AUC were 0.71, 0.92 and 0.88, respectively. Among the various methods and assays, Epipro Colon 2.0 with 2/3 algorithm was the most effective in colorectal cancer detection. Positive ratio of mSEPT9 was higher in advanced CRC (45% in I, 70% in II, 76% in III, 79% in IV) and lower differentiation (31% in high, 73% in moderate, 90% in low) tissue. However, this marker has poor ability of identifying precancerous lesions according to current evidence. Conclusions: mSEPT9 is a reliable blood-based marker in CRC detection, particularly advanced CRC. Epipro Colon 2.0 with 2/3 algorithm is currently the optimal method and assay to detect CRC. PMID:28102859

  1. Molecular methods for the detection of human papillomavirus infection: new insights into their role in diagnostics and epidemiological surveillance

    Directory of Open Access Journals (Sweden)

    Andrea Piana

    2009-06-01

    Full Text Available Human papillomaviruses (HPVs comprise more than 180 genotypes. HPV infection is mainly diagnosed by molecular methods. The aim of our study was to review the main molecular methods used to diagnose HPV infection, underscoring their characteristics. Several methods have been developed for molecular diagnosis of Papilloma infection, such as those based on PCR technique. Another commercial non-PCR based diagnostic method is Hybrid Capture test; it is the only commercially available HPV DNA detection test approved by the FDA. Several Authors have suggested that viral load and E6/E7 transcripts could be used as surrogate markers of persistent HPV infection, being more specific predictors of progressive disease than the simple presence of HPV DNA. Validating clinical sensitivity and specificity of each technique and improving the interpretation of the results are essential; consequently, there is a clear need for well characterized international quality control panels to compare the various diagnostic methods. HPV DNA testing could be useful both as a primary screening test, alone or in combination with a Pap smear, for the early detection of cervical cancer precursors, and as triage test to select women with minor cytological abnormalities who will need further follow-up and to predict possible treatment failure in women with diagnosed high-grade intraepithelial lesions who have undergone excisional therapy. In the next future surveillance for HPV infections, based on these molecular methods, could represent an important step for the development of primary and secondary prophylactic interventions, such as new vaccines targeted to genotypes who might replace those previously prevalent.

  2. Novel portable platform for molecular detection of toxigenic Clostridium difficile in faeces: a diagnostic accuracy study.

    Science.gov (United States)

    Hirvonen, J J; Matero, P; Siebert, C; Kauppila, J; Vuento, R; Tuokko, H; Boisset, S

    2017-05-01

    A novel portable platform for nucleic acid amplification enables rapid detection of diarrhoea causing toxigenic Clostridium difficile directly from faeces, even in resource-limited settings. We evaluated the accuracy and precision of the new commercial molecular test system. One thousand one hundred and sixty faecal samples from patients suspected of having Clostridium difficile infection (CDI) were analysed using the Orion GenRead C. difficile test system (Orion Diagnostica Oy, Espoo, Finland) and comparative methods in three teaching hospital laboratories in Finland and France. The precision of the Orion GenRead C. difficile test system was evaluated in a reproducibility study with a set of blind-coded samples. The test system is based on a new isothermal amplification technology (Strand Invasion Based Amplification, SIBA®) and detection of the tcdB gene of C. difficile. We calculated the sensitivity, specificity, and the overall agreement according to Clinical and Laboratory Standards Institute recommendations. The overall agreement of the Orion GenRead C. difficile test when compared to the comparative methods in routine use in the participating laboratories was between 96.7% and 98.8%. In the reproducibility study; the total percent agreement between three laboratories was 99.8%. The identification of toxigenic C. difficile from faeces with the light-weight portable Orion GenRead test system was highly sensitive and specific, and the results were reproducible in the participating laboratories. This platform could enable fast and accurate molecular pathogen detection even in resource-limited or point-of-care settings.

  3. Automated Micro-Object Detection for Mobile Diagnostics Using Lens-Free Imaging Technology

    Directory of Open Access Journals (Sweden)

    Mohendra Roy

    2016-05-01

    Full Text Available Lens-free imaging technology has been extensively used recently for microparticle and biological cell analysis because of its high throughput, low cost, and simple and compact arrangement. However, this technology still lacks a dedicated and automated detection system. In this paper, we describe a custom-developed automated micro-object detection method for a lens-free imaging system. In our previous work (Roy et al., we developed a lens-free imaging system using low-cost components. This system was used to generate and capture the diffraction patterns of micro-objects and a global threshold was used to locate the diffraction patterns. In this work we used the same setup to develop an improved automated detection and analysis algorithm based on adaptive threshold and clustering of signals. For this purpose images from the lens-free system were then used to understand the features and characteristics of the diffraction patterns of several types of samples. On the basis of this information, we custom-developed an automated algorithm for the lens-free imaging system. Next, all the lens-free images were processed using this custom-developed automated algorithm. The performance of this approach was evaluated by comparing the counting results with standard optical microscope results. We evaluated the counting results for polystyrene microbeads, red blood cells, and HepG2, HeLa, and MCF7 cells. The comparison shows good agreement between the systems, with a correlation coefficient of 0.91 and linearity slope of 0.877. We also evaluated the automated size profiles of the microparticle samples. This Wi-Fi-enabled lens-free imaging system, along with the dedicated software, possesses great potential for telemedicine applications in resource-limited settings.

  4. [Neuroimaging technique: a diagnostic tool to detect altered states of consciousness].

    Science.gov (United States)

    Thonnard, Marie; Boly, Mélanie; Bruno, Marie-Aurélie; Chatelle, Camille; Gosseries, Olivia; Laureys, Steven; Vanhaudenhuyse, Audrey

    2011-01-01

    Vegetative and minimally conscious states diagnosis remained a major clinical challenge. New paradigms such as measurement of the global cerebral metabolism, the structural and functional integrity of fronto-parietal network, or the spontaneous activity in resting state have been shown to be helpful to disentangle vegetative from minimally conscious patients. Active neuroimagery paradigms also allow detecting voluntary and conscious activity in non-communicative patients. The implementation of these methods in clinical routine could permit to reduce the current high rate of misdiagnosis (40%).

  5. Multi-chamber and multi-layer thiol-ene microchip for cell culture

    DEFF Research Database (Denmark)

    Tan, H. Y.; Hemmingsen, Mette; Lafleur, Josiane P.

    2014-01-01

    We present a multi-layer and multi-chamber microfluidic chip fabricated using two different thiol-ene mixtures. Sandwiched between the thiol-ene chip layers is a commercially available membrane whose morphology has been altered with coatings of thiol-ene mixtures. Experiments have been conducted ...... with the microchip and shown that the fabricated microchip is suitable for long term cell culture....

  6. Detecting an elusive invasive species: a diagnostic PCR to detect Burmese python in Florida waters and an assessment of persistence of environmental DNA.

    Science.gov (United States)

    Piaggio, Antoinette J; Engeman, Richard M; Hopken, Matthew W; Humphrey, John S; Keacher, Kandy L; Bruce, William E; Avery, Michael L

    2014-03-01

    Recent studies have demonstrated that detection of environmental DNA (eDNA) from aquatic vertebrates in water bodies is possible. The Burmese python, Python bivittatus, is a semi-aquatic, invasive species in Florida where its elusive nature and cryptic coloration make its detection difficult. Our goal was to develop a diagnostic PCR to detect P. bivittatus from water-borne eDNA, which could assist managers in monitoring this invasive species. First, we used captive P. bivittatus to determine whether reptilian DNA could be isolated and amplified from water samples. We also evaluated the efficacy of two DNA isolation methods and two DNA extraction kits commonly used in eDNA preparation. A fragment of the mitochondrial cytochrome b gene from P. bivittatus was detected in all water samples isolated with the sodium acetate precipitate and the QIAamp DNA Micro Kit. Next, we designed P. bivittatus-specific primers and assessed the degradation rate of eDNA in water. Our primers did not amplify DNA from closely related species, and we found that P. bivittatus DNA was consistently detectable up to 96 h. Finally, we sampled water from six field sites in south Florida. Samples from five sites, where P. bivittatus has been observed, tested positive for eDNA. The final site was negative and had no prior documented evidence of P. bivittatus. This study shows P. bivittatus eDNA can be isolated from water samples; thus, this method is a new and promising technique for the management of invasive reptiles.

  7. Comparison of different diagnostic techniques for the detection of cryptosporidiosis in bovines

    Directory of Open Access Journals (Sweden)

    H. K. M. Rekha

    2016-02-01

    Full Text Available Aim: Aim of the present study was to compare different methods, viz., Sheather’s sugar flotation (SSF, Ziehl-Neelsen (ZN, Kinyoun’s acid-fast method (KAF, safranin-methylene blue staining (SMB, and negative staining techniques such as nigrosin staining, light green staining, and malachite green staining for the detection of Cryptosporidium spp. oocysts in bovines. Materials and Methods: A total of 455 fecal samples from bovines were collected from private, government farms and from the clinical cases presented to Department of Medicine, Veterinary College, Bengaluru. They were subjected for SSF, ZN, KAF, SMB and negative staining methods. Results: Out of 455 animal fecal samples screened 5.71% were found positive for Cryptosporidium spp. oocysts. The species were identified as Cryptosporidium parvum in calves and Cryptosporidium andersoni in adults based on the morphological characterization and micrometry of the oocysts. Conclusions: Of all the techniques, fecal flotation with sheather’s was found to be more specific and sensitive method for the detection of Cryptosporidium spp. oocysts. Among the conventional staining methods, the SMB gives better differentiation between oocysts and yeast. Among the three negative staining methods, malachite green was found sensitive over the other methods.

  8. Outlier detection and removal improves accuracy of machine learning approach to multispectral burn diagnostic imaging

    Science.gov (United States)

    Li, Weizhi; Mo, Weirong; Zhang, Xu; Squiers, John J.; Lu, Yang; Sellke, Eric W.; Fan, Wensheng; DiMaio, J. Michael; Thatcher, Jeffrey E.

    2015-12-01

    Multispectral imaging (MSI) was implemented to develop a burn tissue classification device to assist burn surgeons in planning and performing debridement surgery. To build a classification model via machine learning, training data accurately representing the burn tissue was needed, but assigning raw MSI data to appropriate tissue classes is prone to error. We hypothesized that removing outliers from the training dataset would improve classification accuracy. A swine burn model was developed to build an MSI training database and study an algorithm's burn tissue classification abilities. After the ground-truth database was generated, we developed a multistage method based on Z-test and univariate analysis to detect and remove outliers from the training dataset. Using 10-fold cross validation, we compared the algorithm's accuracy when trained with and without the presence of outliers. The outlier detection and removal method reduced the variance of the training data. Test accuracy was improved from 63% to 76%, matching the accuracy of clinical judgment of expert burn surgeons, the current gold standard in burn injury assessment. Given that there are few surgeons and facilities specializing in burn care, this technology may improve the standard of burn care for patients without access to specialized facilities.

  9. DIAGNOSTIC ACCURACY OF VARIOUS METHODS TO DETECT LYMPH NODE METASTASES IN ORAL SQUAMOUS CELL CARCINOMA

    Directory of Open Access Journals (Sweden)

    Priyanka

    2014-05-01

    Full Text Available The present study was undertaken with the purpose to compare the sensitivity of various methods for detection of lymph node metastases:Intra-operative frozen sections H & E staining, Conventional H & E staining on formalin fixed tissue, Serial –step sectioning by conventional H&E staining & Immunohistochemical staining by Pancytokeratin antibody. METHOD: The study included 80 consecutive cases of oral squamous cell carcinoma, who underwent radical neck dissection. The various level of lymph nodes in these cases were checked for metastases by 4 techniques i.e. intra-operative frozen sections H&E staining, conventional H&E staining on formalin fixed tissue, serial –step sectioning by conventional H & E staining &immunohistochemical staining by Pancytokeratin antibody. RESULTS: Considering IHC as a gold standard, we observed highest sensitivity & specificity for serial sectioning at 53.7%, & 98.9% when compared to intraoperative frozen section and conventional H&E which were 32.5%, 97.1% & 44.7%, 98.2% respectively. CONCLUSION: Thus we conclude that the most sensitive method to detect lymph node metastasis in case of Oral Squamous cell carcinoma is step serial section when considering IHC as a gold standard.

  10. Diagnostic agreement between radiologists, dentists and dental students for radiographic detection of approximal caries.

    Directory of Open Access Journals (Sweden)

    Sergio E Uribe

    2015-06-01

    Full Text Available Aim: To assess intraobserver and interobserver agreement for radiographic detection of approximal caries among radiologists, dentists and senior dental students. Materials and methods: 75 standardized bitewing radiographs were examined by four maxillofacial radiologists, four dentists and four senior dental students. Iinterobserver agreement was calculated using a weighted Kappa Cohen test. Two weeks later, the examiners re-evaluated 10% of the sample under the same conditions and intraobserver agreement weighted Kappa Cohen test was calculated. Results: The interobserver Kappa value was 0.68 (good for the dentist-student pair, 0.51 (moderate for the student-radiologist and 0.62 (good for the dentist-radiologist pair. All these differences were significant. The intraobserver agreement Kappa values obtained were 0.56 (moderate for students (p=0.46, 0.46 (moderate for dentists (p<0.05 and 0.68 (good for radiologists. Conclusion: The agreement for proximal caries detection by dental students, dentists and radiologists was moderate to good.

  11. Outlier detection and removal improves accuracy of machine learning approach to multispectral burn diagnostic imaging.

    Science.gov (United States)

    Li, Weizhi; Mo, Weirong; Zhang, Xu; Squiers, John J; Lu, Yang; Sellke, Eric W; Fan, Wensheng; DiMaio, J Michael; Thatcher, Jeffrey E

    2015-12-01

    Multispectral imaging (MSI) was implemented to develop a burn tissue classification device to assist burn surgeons in planning and performing debridement surgery. To build a classification model via machine learning, training data accurately representing the burn tissue was needed, but assigning raw MSI data to appropriate tissue classes is prone to error. We hypothesized that removing outliers from the training dataset would improve classification accuracy. A swine burn model was developed to build an MSI training database and study an algorithm’s burn tissue classification abilities. After the ground-truth database was generated, we developed a multistage method based on Z -test and univariate analysis to detect and remove outliers from the training dataset. Using 10-fold cross validation, we compared the algorithm’s accuracy when trained with and without the presence of outliers. The outlier detection and removal method reduced the variance of the training data. Test accuracy was improved from 63% to 76%, matching the accuracy of clinical judgment of expert burn surgeons, the current gold standard in burn injury assessment. Given that there are few surgeons and facilities specializing in burn care, this technology may improve the standard of burn care for patients without access to specialized facilities.

  12. Methods for improved selectivity in photo-activation and detection of molecular diagnostic agents

    Energy Technology Data Exchange (ETDEWEB)

    Wachter, Eric A. (Oak Ridge, TN); Fisher, Walter G. (Knoxville, TN); Dees, H. Craig (Knoxville, TN)

    2008-03-18

    A method for the imaging of a particular volume of plant or animal tissue, wherein the plant or animal tissue contains at least one photo-active molecular agent. The method comprises the steps of treating the particular volume of the plant or animal tissue with light sufficient to promote a simultaneous two-photon excitation of the photo-active molecular agent contained in the particular volume of the plant or animal tissue, photo-activating at least one of the at least one photo-active molecular agent in the particular volume of the plant or animal tissue, thereby producing at least one photo-activated molecular agent, wherein the at least one photo-activated molecular agent emits energy, detecting the energy emitted by the at least one photo-activated molecular agent, and