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Sample records for rapid real-time detection

  1. Original Article. Evaluation of Rapid Detection of Nasopharyngeal Colonization with MRSA by Real-Time PCR

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    Kang Feng-feng

    2012-03-01

    Full Text Available Objective To investigate the clinical application of Real-Time PCR for rapid detection of methicillin-resistant Staphylococcus aureus (MRSA directly from nasopharyngeal swab specimens.

  2. Rapid quantitative detection of Lactobacillus sakei in meat and fermented sausages by real-time PCR.

    Science.gov (United States)

    Martín, Belén; Jofré, Anna; Garriga, Margarita; Pla, Maria; Aymerich, Teresa

    2006-09-01

    A quick and simple method for quantitative detection of Lactobacillus sakei in fermented sausages was successfully developed. It is based on Chelex-100-based DNA purification and real-time PCR enumeration using a TaqMan fluorescence probe. Primers and probes were designed in the L. sakei 16S-23S rRNA intergenic transcribed spacer region, and the assay was evaluated using L. sakei genomic DNA and an artificially inoculated sausage model. The detection limit of this technique was approximately 3 cells per reaction mixture using both purified DNA and the inoculated sausage model. The quantification limit was established at 30 cells per reaction mixture in both models. The assay was then applied to enumerate L. sakei in real samples, and the results were compared to the MRS agar count method followed by confirmation of the percentage of L. sakei colonies. The results obtained by real-time PCR were not statistically significantly different than those obtained by plate count on MRS agar (P > 0.05), showing a satisfactory agreement between both methods. Therefore, the real-time PCR assay developed can be considered a promising rapid alternative method for the quantification of L. sakei and evaluation of the implantation of starter strains of L. sakei in fermented sausages.

  3. Early Flood Detection for Rapid Humanitarian Response: Harnessing Near Real-Time Satellite and Twitter Signals

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    Brenden Jongman

    2015-10-01

    Full Text Available Humanitarian organizations have a crucial role in response and relief efforts after floods. The effectiveness of disaster response is contingent on accurate and timely information regarding the location, timing and impacts of the event. Here we show how two near-real-time data sources, satellite observations of water coverage and flood-related social media activity from Twitter, can be used to support rapid disaster response, using case-studies in the Philippines and Pakistan. For these countries we analyze information from disaster response organizations, the Global Flood Detection System (GFDS satellite flood signal, and flood-related Twitter activity analysis. The results demonstrate that these sources of near-real-time information can be used to gain a quicker understanding of the location, the timing, as well as the causes and impacts of floods. In terms of location, we produce daily impact maps based on both satellite information and social media, which can dynamically and rapidly outline the affected area during a disaster. In terms of timing, the results show that GFDS and/or Twitter signals flagging ongoing or upcoming flooding are regularly available one to several days before the event was reported to humanitarian organizations. In terms of event understanding, we show that both GFDS and social media can be used to detect and understand unexpected or controversial flood events, for example due to the sudden opening of hydropower dams or the breaching of flood protection. The performance of the GFDS and Twitter data for early detection and location mapping is mixed, depending on specific hydrological circumstances (GFDS and social media penetration (Twitter. Further research is needed to improve the interpretation of the GFDS signal in different situations, and to improve the pre-processing of social media data for operational use.

  4. A one-step, real-time PCR assay for rapid detection of rhinovirus.

    Science.gov (United States)

    Do, Duc H; Laus, Stella; Leber, Amy; Marcon, Mario J; Jordan, Jeanne A; Martin, Judith M; Wadowsky, Robert M

    2010-01-01

    One-step, real-time PCR assays for rhinovirus have been developed for a limited number of PCR amplification platforms and chemistries, and some exhibit cross-reactivity with genetically similar enteroviruses. We developed a one-step, real-time PCR assay for rhinovirus by using a sequence detection system (Applied Biosystems; Foster City, CA). The primers were designed to amplify a 120-base target in the noncoding region of picornavirus RNA, and a TaqMan (Applied Biosystems) degenerate probe was designed for the specific detection of rhinovirus amplicons. The PCR assay had no cross-reactivity with a panel of 76 nontarget nucleic acids, which included RNAs from 43 enterovirus strains. Excellent lower limits of detection relative to viral culture were observed for the PCR assay by using 38 of 40 rhinovirus reference strains representing different serotypes, which could reproducibly detect rhinovirus serotype 2 in viral transport medium containing 10 to 10,000 TCID(50) (50% tissue culture infectious dose endpoint) units/ml of the virus. However, for rhinovirus serotypes 59 and 69, the PCR assay was less sensitive than culture. Testing of 48 clinical specimens from children with cold-like illnesses for rhinovirus by the PCR and culture assays yielded detection rates of 16.7% and 6.3%, respectively. For a batch of 10 specimens, the entire assay was completed in 4.5 hours. This real-time PCR assay enables detection of many rhinovirus serotypes with the Applied Biosystems reagent-instrument platform.

  5. Standardization and application of real-time polymerase chain reaction for rapid detection of bluetongue virus

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    I. Karthika Lakshmi

    2018-04-01

    Full Text Available Aim: The present study was designed to standardize real-time polymerase chain reaction (PCR for detecting the bluetongue virus from blood samples of sheep collected during outbreaks of bluetongue disease in the year 2014 in Andhra Pradesh and Telangana states of India. Materials and Methods: A 10-fold serial dilution of Plasmid PUC59 with bluetongue virus (BTV NS3 insert was used to plot the standard curve. BHK-21 and KC cells were used for in vitro propagation of virus BTV-9 at a TCID50/ml of 105 ml and RNA was isolated by the Trizol method. Both reverse transcription -PCR and real-time PCR using TaqMan probe were carried out with RNA extracted from virus-spiked culture medium and blood to compare the sensitivity by means of finding out the limit of detection (LoD. The results were verified by inoculating the detected and undetected dilutions onto cell cultures with further cytological (cytopathic effect and molecular confirmation (by BTV-NS1 group-specific PCR. The standardized technique was then applied to field samples (blood for detecting BTV. Results: The slope of the standard curve obtained was -3.23, and the efficiency was 103%. The LoD with RT-PCR was 8.269Ex103 number of copies of plasmid, whereas it was 13 with real-time PCR for plasmid dilutions. Similarly, LoD was determined for virus-spiked culture medium, and blood with both the types of PCR and the values were 103 TCID 50/ml and 104 TCID 50/ml with RT-PCR and 10° TCID 50/ml and 102 TCID 50/ml with real-time PCR, respectively. The standardized technique was applied to blood samples collected from BTV suspected animals; 10 among 20 samples were found positive with Cq values ranging from 27 to 39. The Cq value exhibiting samples were further processed in cell cultures and were confirmed to be BT positive. Likewise, Cq undetected samples on processing in cell cultures turned out to be BTV negative. Conclusion: Real-time PCR was found to be a very sensitive as well as reliable method

  6. Rapid detection of Van genes in rectal swabs by real time PCR in Southern Brazil

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    Vlademir Cantarelli

    2011-10-01

    Full Text Available INTRODUCTION: Laboratory-based surveillance is an important component in the control of vancomycin resistant enterococci (VRE. METHODS: The study aimed to evaluate real-time polymerase chain reaction (RT-PCR (genes vanA-vanB for VRE detection on 115 swabs from patients included in a surveillance program. RESULTS: Sensitivity of RT-PCR was similar to primary culture (75% and 79.5%, respectively when compared to broth enriched culture, whereas specificity was 83.1%. CONCLUSIONS: RT-PCR provides same day results, however it showed low sensitivity for VRE detection.

  7. Development and Validation of a Real-Time PCR Assay for Rapid Detection of Candida auris from Surveillance Samples.

    Science.gov (United States)

    Leach, L; Zhu, Y; Chaturvedi, S

    2018-02-01

    Candida auris is an emerging multidrug-resistant yeast causing invasive health care-associated infection with high mortality worldwide. Rapid identification of C. auris is of primary importance for the implementation of public health measures to control the spread of infection. To achieve these goals, we developed and validated a TaqMan-based real-time PCR assay targeting the internal transcribed spacer 2 ( ITS 2) region of the ribosomal gene. The assay was highly specific, reproducible, and sensitive, with the detection limit of 1 C. auris CFU/PCR. The performance of the C. auris real-time PCR assay was evaluated by using 623 surveillance samples, including 365 patient swabs and 258 environmental sponges. Real-time PCR yielded positive results from 49 swab and 58 sponge samples, with 89% and 100% clinical sensitivity with regard to their respective culture-positive results. The real-time PCR also detected C. auris DNA from 1% and 12% of swab and sponge samples with culture-negative results, indicating the presence of dead or culture-impaired C. auris The real-time PCR yielded results within 4 h of sample processing, compared to 4 to 14 days for culture, reducing turnaround time significantly. The new real-time PCR assay allows for accurate and rapid screening of C. auris and can increase effective control and prevention of this emerging multidrug-resistant fungal pathogen in health care facilities. Copyright © 2018 Leach et al.

  8. Rapid detection of Salmonella in pet food: design and evaluation of integrated methods based on real-time PCR detection.

    Science.gov (United States)

    Balachandran, Priya; Friberg, Maria; Vanlandingham, V; Kozak, K; Manolis, Amanda; Brevnov, Maxim; Crowley, Erin; Bird, Patrick; Goins, David; Furtado, Manohar R; Petrauskene, Olga V; Tebbs, Robert S; Charbonneau, Duane

    2012-02-01

    Reducing the risk of Salmonella contamination in pet food is critical for both companion animals and humans, and its importance is reflected by the substantial increase in the demand for pathogen testing. Accurate and rapid detection of foodborne pathogens improves food safety, protects the public health, and benefits food producers by assuring product quality while facilitating product release in a timely manner. Traditional culture-based methods for Salmonella screening are laborious and can take 5 to 7 days to obtain definitive results. In this study, we developed two methods for the detection of low levels of Salmonella in pet food using real-time PCR: (i) detection of Salmonella in 25 g of dried pet food in less than 14 h with an automated magnetic bead-based nucleic acid extraction method and (ii) detection of Salmonella in 375 g of composite dry pet food matrix in less than 24 h with a manual centrifugation-based nucleic acid preparation method. Both methods included a preclarification step using a novel protocol that removes food matrix-associated debris and PCR inhibitors and improves the sensitivity of detection. Validation studies revealed no significant differences between the two real-time PCR methods and the standard U.S. Food and Drug Administration Bacteriological Analytical Manual (chapter 5) culture confirmation method.

  9. Development and validation of real-time PCR for rapid detection of Mecistocirrus digitatus.

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    Subhra Subhadra

    Full Text Available Hematophagous activity of Mecistocirrus digitatus, which causes substantial blood and weight loss in large ruminants, is an emerging challenge due to the economic loss it brings to the livestock industry. Infected animals are treated with anthelmintic drugs, based on the identification of helminth species and the severity of infection; however, traditional methods such as microscopic identification and the counting of eggs for diagnosis and determination of level of infection are laborious, cumbersome and unreliable. To facilitate the detection of this parasite, a SYBR green-based real-time PCR was standardized and validated for the detection of M. digitatus infection in cattle and buffaloes. Oligonucleotides were designed to amplify partial Internal Transcribed Spacer (ITS-1 sequence of M. digitatus. The specificity of the primers was confirmed by non-amplification of DNA extracted from other commonly occurring gastrointestinal nematodes in ruminants. Plasmids were ligated with partial ITS-1 sequence of M. digitatus, serially diluted (hundred fold and used as standards in the real-time PCR assay. The quantification cycle (Cq values were plotted against the standard DNA concentration to produce a standard curve. The assay was sensitive enough to detect one plasmid containing the M. digitatus DNA. Clinical application of this assay was validated by testing the DNA extracted from the faeces of naturally infected cattle (n = 40 and buffaloes (n = 25. The results were compared with our standard curve to calculate the quantity of M. digitatus in each faecal sample. The Cq value of the assay depicted a strong linear relationship with faecal DNA content, with a regression coefficient of 0.984 and efficiency of 99%. This assay has noteworthy advantages over the conventional methods of diagnosis because it is more specific, sensitive and reliable.

  10. Rapid Detection and Differentiation of Clonorchis sinensis and Opisthorchis viverrini Using Real-Time PCR and High Resolution Melting Analysis

    OpenAIRE

    Cai, Xian-Quan; Yu, Hai-Qiong; Li, Rong; Yue, Qiao-Yun; Liu, Guo-Hua; Bai, Jian-Shan; Deng, Yan; Qiu, De-Yi; Zhu, Xing-Quan

    2014-01-01

    Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA...

  11. Rapid and real-time detection technologies for emerging viruses of ...

    Indian Academy of Sciences (India)

    2008-10-17

    Oct 17, 2008 ... The development of technologies with rapid and sensitive detection capabilities and increased throughput have become crucial for responding to greater number threats posed by emerging and re-emerging viruses in the recent past. The conventional identification methods require time-consuming culturing ...

  12. Rapid and sensitive detection of canine distemper virus by real-time reverse transcription recombinase polymerase amplification.

    Science.gov (United States)

    Wang, Jianchang; Wang, Jinfeng; Li, Ruiwen; Liu, Libing; Yuan, Wanzhe

    2017-08-15

    Canine distemper, caused by Canine distemper virus (CDV), is a highly contagious and fatal systemic disease in free-living and captive carnivores worldwide. Recombinase polymerase amplification (RPA), as an isothermal gene amplification technique, has been explored for the molecular detection of diverse pathogens. A real-time reverse transcription RPA (RT-RPA) assay for the detection of canine distemper virus (CDV) using primers and exo probe targeting the CDV nucleocapsid protein gene was developed. A series of other viruses were tested by the RT-RPA.Thirty-two field samples were further tested by RT-RPA, and the resuts were compared with those obtained by the real-time RT-PCR. The RT-RPA assay was performed successfully at 40 °C, and the results were obtained within 3 min-12 min. The assay could detect CDV, but did not show cross-detection of canine parvovirus-2 (CPV-2), canine coronavirus (CCoV), canine parainfluenza virus (CPIV), pseudorabies virus (PRV) or Newcastle disease virus (NDV), demonstrating high specificity. The analytical sensitivity of RT-RPA was 31.8 copies in vitro transcribed CDV RNA, which is 10 times lower than the real-time RT-PCR. The assay performance was validated by testing 32 field samples and compared to real-time RT-PCR. The results indicated an excellent correlation between RT-RPA and a reference real-time RT-PCR method. Both assays provided the same results, and R 2 value of the positive results was 0.947. The results demonstrated that the RT-RPA assay offers an alternative tool for simple, rapid, and reliable detection of CDV both in the laboratory and point-of-care facility, especially in the resource-limited settings.

  13. Real-time PCR-based method for rapid detection of Aspergillus niger and Aspergillus welwitschiae isolated from coffee.

    Science.gov (United States)

    von Hertwig, Aline Morgan; Sant'Ana, Anderson S; Sartori, Daniele; da Silva, Josué José; Nascimento, Maristela S; Iamanaka, Beatriz Thie; Pelegrinelli Fungaro, Maria Helena; Taniwaki, Marta Hiromi

    2018-05-01

    Some species from Aspergillus section Nigri are morphologically very similar and altogether have been called A. niger aggregate. Although the species included in this group are morphologically very similar, they differ in their ability to produce mycotoxins and other metabolites and their taxonomical status has evolved continuously. Among them, A. niger and A. welwitschiae are ochratoxin A and fumonisin B 2 producers and their detection and/or identification is of crucial importance for food safety. The aim of this study was the development of a real-time PCR-based method for simultaneous discrimination of A. niger and A. welwitschiae from other species of the A. niger aggregate isolated from coffee beans. One primer pair and a hybridization probe specific for detection of A. niger and A. welwitschiae strains were designed based on the BenA gene sequences, and used in a Real-time PCR assay for the rapid discrimination between both these species from all others of the A. niger aggregate. The Real-time PCR assay was shown to be 100% efficient in discriminating the 73 isolates of A. niger/A. welwitschiae from the other A. niger aggregate species analyzed as a negative control. This result testifies to the use of this technique as a good tool in the rapid detection of these important toxigenic species. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. A real-time loop-mediated isothermal amplification assay for rapid detection of Shigella species.

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    Liew, P S; Teh, C S J; Lau, Y L; Thong, K L

    2014-12-01

    Shigellosis is a foodborne illness caused by the genus Shigella and is an important global health issue. The development of effective techniques for rapid detection of this pathogen is essential for breaking the chain of transmission. Therefore, we have developed a novel loop-mediated isothermal amplification (LAMP) assay targeting the invasion plasmid antigen H (ipaH) gene to rapidly detect Shigella species. This assay could be performed in 90 min at an optimal temperature of 64ºC, with endpoint results visualized directly. Notably, the method was found to be more sensitive than conventional PCR. Indeed, the detection limit for the LAMP assay on pure bacterial cultures was 5.9 x 10(5) CFU/ml, while PCR displayed a limit of 5.9 x 10(7) CFU/ml. In spiked lettuce samples, the sensitivity of the LAMP assay was 3.6 x 10(4) CFU/g, whereas PCR was 3.6 x 10(5) CFU/g. Overall, the assay accurately identified 32 Shigella spp. with one enteroinvasive Escherichia coli displaying positive reaction while the remaining 32 non-Shigella strains tested were negative.

  15. Rapid detection and typing of pathogenic nonpneumophila Legionella spp. isolates using a multiplex real-time PCR assay.

    Science.gov (United States)

    Benitez, Alvaro J; Winchell, Jonas M

    2016-04-01

    We developed a single tube multiplex real-time PCR assay that allows for the rapid detection and typing of 9 nonpneumophila Legionella spp. isolates that are clinically relevant. The multiplex assay is capable of simultaneously detecting and discriminating L. micdadei, L. bozemanii, L. dumoffii, L. longbeachae, L. feeleii, L. anisa, L. parisiensis, L. tucsonensis serogroup (sg) 1 and 3, and L. sainthelensis sg 1 and 2 isolates. Evaluation of the assay with nucleic acid from each of these species derived from both clinical and environmental isolates and typing strains demonstrated 100% sensitivity and 100% specificity when tested against 43 other Legionella spp. Typing of L. anisa, L. parisiensis, and L. tucsonensis sg 1 and 3 isolates was accomplished by developing a real-time PCR assay followed by high-resolution melt (HRM) analysis targeting the ssrA gene. Further typing of L. bozemanii, L. longbeachae, and L. feeleii isolates to the serogroup level was accomplished by developing a real-time PCR assay followed by HRM analysis targeting the mip gene. When used in conjunction with other currently available diagnostic tests, these assays may aid in rapidly identifying specific etiologies associated with Legionella outbreaks, clusters, sporadic cases, and potential environmental sources. Published by Elsevier Inc.

  16. Micropower Impulse Radar: A Novel Technology for Rapid, Real-Time Detection of Pneumothorax

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    Phillip D. Levy

    2011-01-01

    Full Text Available Pneumothorax detection in emergency situations must be rapid and at the point of care. Current standards for detection of a pneumothorax are supine chest X-rays, ultrasound, and CT scans. Unfortunately these tools and the personnel necessary for their facile utilization may not be readily available in acute circumstances, particularly those which occur in the pre-hospital setting. The decision to treat therefore, is often made without adequate information. In this report, we describe a novel hand-held device that utilizes Micropower Impulse Radar to reliably detect the presence of a pneumothorax. The technology employs ultra wide band pulses over a frequency range of 500 MHz to 6 GHz and a proprietary algorithm analyzes return echoes to determine if a pneumothorax is present with no user interpretation required. The device has been evaluated in both trauma and surgical environments with sensitivity of 93% and specificity of 85%. It is has the CE Mark and is available for sale in Europe. Post market studies are planned starting in May of 2011. Clinical studies to support the FDA submission will be completed in the first quarter of 2012.

  17. TaqMan MGB probe fluorescence real-time quantitative PCR for rapid detection of Chinese Sacbrood virus.

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    Ma Mingxiao

    Full Text Available Sacbrood virus (SBV is a picorna-like virus that affects honey bees (Apis mellifera and results in the death of the larvae. Several procedures are available to detect Chinese SBV (CSBV in clinical samples, but not to estimate the level of CSBV infection. The aim of this study was develop an assay for rapid detection and quantification of this virus. Primers and probes were designed that were specific for CSBV structural protein genes. A TaqMan minor groove binder (MGB probe-based, fluorescence real-time quantitative PCR was established. The specificity, sensitivity and stability of the assay were assessed; specificity was high and there were no cross-reactivity with healthy larvae or other bee viruses. The assay was applied to detect CSBV in 37 clinical samples and its efficiency was compared with clinical diagnosis, electron microscopy observation, and conventional RT-PCR. The TaqMan MGB-based probe fluorescence real-time quantitative PCR for CSBV was more sensitive than other methods tested. This assay was a reliable, fast, and sensitive method that was used successfully to detect CSBV in clinical samples. The technology can provide a useful tool for rapid detection of CSBV. This study has established a useful protocol for CSBV testing, epidemiological investigation, and development of animal models.

  18. Rapid detection of Lactobacillus kefiranofaciens in kefir grain and kefir milk using newly developed real-time PCR.

    Science.gov (United States)

    Kim, Dong-Hyeon; Chon, Jung-Whan; Kim, Hong-Seok; Yim, Jin-Hyeok; Kim, Hyunsook; Seo, Kun-Ho

    2015-04-01

    Lactobacillus kefiranofaciens is an indicator microorganism for kefir and a key factor in kefir grain formation and kefiran production. We designed a novel real-time PCR primer and probe set, LKF_KU504, for the rapid detection of L. kefiranofaciens. In inclusivity and exclusivity tests, only 14 L. kefiranofaciens strains were positive among 61 microorganisms, indicating 100 % sensitivity and specificity. The LKF_KU504 set also differentiated kefir milk from 30 commercial nonkefir yogurts. The levels of L. kefiranofaciens in kefir grain and kefir milk were significantly different, indicating L. kefiranofaciens was more concentrated in kefir grain than in kefir milk.

  19. Real time freeway incident detection.

    Science.gov (United States)

    2014-04-01

    The US Department of Transportation (US-DOT) estimates that over half of all congestion : events are caused by highway incidents rather than by rush-hour traffic in big cities. Real-time : incident detection on freeways is an important part of any mo...

  20. Rapid detection of human parechoviruses in clinical samples by real-time PCR

    NARCIS (Netherlands)

    Benschop, Kimberley; Molenkamp, Richard; van der Ham, Alwin; Wolthers, Katja; Beld, Marcel

    2008-01-01

    BACKGROUND: Human parechoviruses (HPeVs) have been associated with severe conditions such as neonatal sepsis and meningitis in young children. Rapid identification of an infectious agent in such serious conditions in these patients is essential for adequate decision making regarding treatment and

  1. Rapid and real-time detection technologies for emerging viruses of ...

    Indian Academy of Sciences (India)

    Prakash

    methods are available which make it possible to detect and analyze any virus, including .... qualitative, or 'yes/no' format. ... out of the pure research laboratory and into the diagnostic .... types A and B, respiratory syncytial virus (RSV) and para.

  2. A rapid detection method for policy-sensitive amines real-time supervision.

    Science.gov (United States)

    Zhang, Haixu; Shu, Jinian; Yang, Bo; Zhang, Peng; Ma, Pengkun

    2018-02-01

    Many organic amines that comprise a benzene ring are policy-sensitive because of their toxicity and links to social harm. However, to date, detection of such compounds mainly relies on offline methods. This study proposes an online pptv (parts per trillion by volume) level of detection method for amines, using the recently-built vacuum ultraviolet photoionization mass spectrometer (VUV-PIMS) combined with a new doping technique. Thus, the dichloromethane doping-assisted photoionization mass spectra of aniline, benzylamine, phenethylamine, amphetamine, and their structural isomers were recorded. The dominant characteristic mass peaks for all amines are those afforded by protonated amines and the amino radical-loss. The signal intensities of the amines were enhanced by 60-130 times compared to those recorded without doping assistance. Under 10s detection time, the sensitivities of aniline and benzylamine in the gas phase were determined as 4.0 and 2.7 countspptv -1 , with limits of detection (LODs) of 36 and 22 pptv, respectively. Notably, the detection efficiency of this method can be tenfold better in future applications since the ion transmission efficiency of the mass spectrometer was intentionally reduced to ~ 10% in this study. Therefore, dichloromethane doping-assisted photoionization mass spectrometry has proven to be a highly promising on-line approach to amine detection in environmental and judicial supervision and shows great potential for application in the biological field. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. A novel photoinduced electron transfer (PET) primer technique for rapid real-time PCR detection of Cryptosporidium spp

    International Nuclear Information System (INIS)

    Jothikumar, N.; Hill, Vincent R.

    2013-01-01

    Highlights: •Uses a single-labeled fluorescent primer for real-time PCR. •The detection sensitivity of PET PCR was comparable to TaqMan PCR. •Melt curve analysis can be performed to confirm target amplicon production. •Conventional PCR primers can be converted to PET PCR primers. -- Abstract: We report the development of a fluorescently labeled oligonucleotide primer that can be used to monitor real-time PCR. The primer has two parts, the 3′-end of the primer is complimentary to the target and a universal 17-mer stem loop at the 5′-end forms a hairpin structure. A fluorescent dye is attached to 5′-end of either the forward or reverse primer. The presence of guanosine residues at the first and second position of the 3′ dangling end effectively quenches the fluorescence due to the photo electron transfer (PET) mechanism. During the synthesis of nucleic acid, the hairpin structure is linearized and the fluorescence of the incorporated primer increases several-fold due to release of the fluorescently labeled tail and the absence of guanosine quenching. As amplicons are synthesized during nucleic acid amplification, the fluorescence increase in the reaction mixture can be measured with commercially available real-time PCR instruments. In addition, a melting procedure can be performed to denature the double-stranded amplicons, thereby generating fluorescence peaks that can differentiate primer dimers and other non-specific amplicons if formed during the reaction. We demonstrated the application of PET-PCR for the rapid detection and quantification of Cryptosporidium parvum DNA. Comparison with a previously published TaqMan® assay demonstrated that the two real-time PCR assays exhibited similar sensitivity for a dynamic range of detection of 6000–0.6 oocysts per reaction. PET PCR primers are simple to design and less-expensive than dual-labeled probe PCR methods, and should be of interest for use by laboratories operating in resource

  4. A novel photoinduced electron transfer (PET) primer technique for rapid real-time PCR detection of Cryptosporidium spp

    Energy Technology Data Exchange (ETDEWEB)

    Jothikumar, N., E-mail: jin2@cdc.gov; Hill, Vincent R.

    2013-06-28

    Highlights: •Uses a single-labeled fluorescent primer for real-time PCR. •The detection sensitivity of PET PCR was comparable to TaqMan PCR. •Melt curve analysis can be performed to confirm target amplicon production. •Conventional PCR primers can be converted to PET PCR primers. -- Abstract: We report the development of a fluorescently labeled oligonucleotide primer that can be used to monitor real-time PCR. The primer has two parts, the 3′-end of the primer is complimentary to the target and a universal 17-mer stem loop at the 5′-end forms a hairpin structure. A fluorescent dye is attached to 5′-end of either the forward or reverse primer. The presence of guanosine residues at the first and second position of the 3′ dangling end effectively quenches the fluorescence due to the photo electron transfer (PET) mechanism. During the synthesis of nucleic acid, the hairpin structure is linearized and the fluorescence of the incorporated primer increases several-fold due to release of the fluorescently labeled tail and the absence of guanosine quenching. As amplicons are synthesized during nucleic acid amplification, the fluorescence increase in the reaction mixture can be measured with commercially available real-time PCR instruments. In addition, a melting procedure can be performed to denature the double-stranded amplicons, thereby generating fluorescence peaks that can differentiate primer dimers and other non-specific amplicons if formed during the reaction. We demonstrated the application of PET-PCR for the rapid detection and quantification of Cryptosporidium parvum DNA. Comparison with a previously published TaqMan® assay demonstrated that the two real-time PCR assays exhibited similar sensitivity for a dynamic range of detection of 6000–0.6 oocysts per reaction. PET PCR primers are simple to design and less-expensive than dual-labeled probe PCR methods, and should be of interest for use by laboratories operating in resource

  5. 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.

  6. Rapid detection of sugar alcohol precursors and corresponding nitrate ester explosives using direct analysis in real time mass spectrometry.

    Science.gov (United States)

    Sisco, Edward; Forbes, Thomas P

    2015-04-21

    This work highlights the rapid detection of nitrate ester explosives and their sugar alcohol precursors by direct analysis in real time mass spectrometry (DART-MS) using an off-axis geometry. Demonstration of the effect of various parameters, such as ion polarity and in-source collision induced dissociation (CID) on the detection of these compounds is presented. Sensitivity of sugar alcohols and nitrate ester explosives was found to be greatest in negative ion mode with sensitivities ranging from hundreds of picograms to hundreds of nanograms, depending on the characteristics of the particular molecule. Altering the in-source CID potential allowed for acquisition of characteristic molecular ion spectra as well as fragmentation spectra. Additional studies were completed to identify the role of different experimental parameters on the sensitivity for these compounds. Variables that were examined included the DART gas stream temperature, the presence of a related compound (i.e., the effect of a precursor on the detection of a nitrate ester explosive), incorporation of dopant species and the role of the analysis surface. It was determined that each variable affected the response and detection of both sugar alcohols and the corresponding nitrate ester explosives. From this work, a rapid and sensitive method for the detection of individual sugar alcohols and corresponding nitrate ester explosives, or mixtures of the two, has been developed, providing a useful tool in the real-world identification of homemade explosives.

  7. Single tube multiplex real-time PCR for the rapid detection of herpesvirus infections of the central nervous system.

    Science.gov (United States)

    Sankuntaw, Nipaporn; Sukprasert, Saovaluk; Engchanil, Chulapan; Kaewkes, Wanlop; Chantratita, Wasun; Pairoj, Vantanit; Lulitanond, Viraphong

    2011-01-01

    Human herpesvirus infection of immunocompromised hosts may lead to central nervous system (CNS) infection and diseases. In this study, a single tube multiplex real-time PCR was developed for the detection of five herpesviruses (HSV-1, HSV-2, VZV, EBV and CMV) in clinical cerebrospinal fluid (CSF) specimens. Two primer pairs specific for the herpesvirus polymerase gene and five hybridization probe pairs for the specific identification of the herpesvirus types were used in a LightCycler multiplex real-time PCR. A singleplex real-time PCR was first optimized and then applied to the multiplex real-time PCR. The singleplex and multiplex real-time PCRs showed no cross-reactivity. The sensitivity of the singleplex real-time PCR was 1 copy per reaction for each herpesvirus, while that of the multiplex real-time PCR was 1 copy per reaction for HSV-1 and VZV and 10 copies per reaction for HSV-2, EBV and CMV. Intra and inter-assay variations of the single tube multiplex assay were in the range of 0.02%-3.67% and 0.79%-4.35%, respectively. The assay was evaluated by testing 62 clinical CSF samples and was found to have equivalent sensitivity, specificity and agreement as the routine real-time PCR, but reducing time, cost and amount of used sample. Copyright © 2011 Elsevier Ltd. All rights reserved.

  8. Real-time PCR-based method for the rapid detection of extended RAS mutations using bridged nucleic acids in colorectal cancer.

    Science.gov (United States)

    Iida, Takao; Mizuno, Yukie; Kaizaki, Yasuharu

    2017-10-27

    Mutations in RAS and BRAF are predictors of the efficacy of anti-epidermal growth factor receptor (EGFR) therapy in patients with metastatic colorectal cancer (mCRC). Therefore, simple, rapid, cost-effective methods to detect these mutations in the clinical setting are greatly needed. In the present study, we evaluated BNA Real-time PCR Mutation Detection Kit Extended RAS (BNA Real-time PCR), a real-time PCR method that uses bridged nucleic acid clamping technology to rapidly detect mutations in RAS exons 2-4 and BRAF exon 15. Genomic DNA was extracted from 54 formalin-fixed paraffin-embedded (FFPE) tissue samples obtained from mCRC patients. Among the 54 FFPE samples, BNA Real-time PCR detected 21 RAS mutations (38.9%) and 5 BRAF mutations (9.3%), and the reference assay (KRAS Mutation Detection Kit and MEBGEN™ RASKET KIT) detected 22 RAS mutations (40.7%). The concordance rate of detected RAS mutations between the BNA Real-time PCR assay and the reference assays was 98.2% (53/54). The BNA Real-time PCR assay proved to be a more simple, rapid, and cost-effective method for detecting KRAS and RAS mutations compared with existing assays. These findings suggest that BNA Real-time PCR is a valuable tool for predicting the efficacy of early anti-EGFR therapy in mCRC patients. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Rapid detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini using real-time PCR and high resolution melting analysis.

    Science.gov (United States)

    Cai, Xian-Quan; Yu, Hai-Qiong; Li, Rong; Yue, Qiao-Yun; Liu, Guo-Hua; Bai, Jian-Shan; Deng, Yan; Qiu, De-Yi; Zhu, Xing-Quan

    2014-01-01

    Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA extracted from the two flukes yielded specific amplification and their identity was confirmed by sequencing, having the accuracy of 100% in reference to conventional methods. The assay was proved to be highly sensitive with a detection limit below 1 pg of purified genomic DNA, 5 EPG, or 1 metacercaria of C. sinensis. Moreover, C. sinensis and O. viverrini were able to be differentiated by their HRM profiles. The method can reduce labor of microscopic examination and the contamination of agarose electrophoresis. Moreover, it can differentiate these two flukes which are difficult to be distinguished using other methods. The established method provides an alternative tool for rapid, simple, and duplex detection of C. sinensis and O. viverrini.

  10. Rapid Detection and Differentiation of Clonorchis sinensis and Opisthorchis viverrini Using Real-Time PCR and High Resolution Melting Analysis

    Directory of Open Access Journals (Sweden)

    Xian-Quan Cai

    2014-01-01

    Full Text Available Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA extracted from the two flukes yielded specific amplification and their identity was confirmed by sequencing, having the accuracy of 100% in reference to conventional methods. The assay was proved to be highly sensitive with a detection limit below 1 pg of purified genomic DNA, 5 EPG, or 1 metacercaria of C. sinensis. Moreover, C. sinensis and O. viverrini were able to be differentiated by their HRM profiles. The method can reduce labor of microscopic examination and the contamination of agarose electrophoresis. Moreover, it can differentiate these two flukes which are difficult to be distinguished using other methods. The established method provides an alternative tool for rapid, simple, and duplex detection of C. sinensis and O. viverrini.

  11. Rapid, Real-time Methane Detection in Ground Water Using a New Gas-Water Equilibrator Design

    Science.gov (United States)

    Ruybal, C. J.; DiGiulio, D. C.; Wilkin, R. T.; Hargrove, K. D.; McCray, J. E.

    2014-12-01

    Recent increases in unconventional gas development have been accompanied by public concern for methane contamination in drinking water wells near production areas. Although not a regulated pollutant, methane may be a marker contaminant for others that are less mobile in groundwater and thus may be detected later, or at a location closer to the source. In addition, methane poses an explosion hazard if exsolved concentrations reach 5 - 15% volume in air. Methods for determining dissolved gases, such as methane, have evolved over 60 years. However, the response time of these methods is insufficient to monitor trends in methane concentration in real-time. To enable rapid, real-time monitoring of aqueous methane concentrations during ground water purging, a new gas-water equilibrator (GWE) was designed that increases gas-water mass exchange rates of methane for measurement. Monitoring of concentration trends allows a comparison of temporal trends between sampling events and comparison of baseline conditions with potential post-impact conditions. These trends may be a result of removal of stored casing water, pre-purge ambient borehole flow, formation physical and chemical heterogeneity, or flow outside of well casing due to inadequate seals. Real-time information in the field can help focus an investigation, aid in determining when to collect a sample, save money by limiting costs (e.g. analytical, sample transport and storage), and provide an immediate assessment of local methane concentrations. Four domestic water wells, one municipal water well, and one agricultural water well were sampled for traditional laboratory analysis and compared to the field GWE results. Aqueous concentrations measured on the GWE ranged from non-detect to 1,470 μg/L methane. Some trends in aqueous methane concentrations measured on the GWE were observed during purging. Applying a paired t-test comparing the new GWE method and traditional laboratory analysis yielded a p-value 0

  12. Rapid prototyping of SoC-based real-time vision system: application to image preprocessing and face detection

    Science.gov (United States)

    Jridi, Maher; Alfalou, Ayman

    2017-05-01

    By this paper, the major goal is to investigate the Multi-CPU/FPGA SoC (System on Chip) design flow and to transfer a know-how and skills to rapidly design embedded real-time vision system. Our aim is to show how the use of these devices can be benefit for system level integration since they make possible simultaneous hardware and software development. We take the facial detection and pretreatments as case study since they have a great potential to be used in several applications such as video surveillance, building access control and criminal identification. The designed system use the Xilinx Zedboard platform. The last is the central element of the developed vision system. The video acquisition is performed using either standard webcam connected to the Zedboard via USB interface or several camera IP devices. The visualization of video content and intermediate results are possible with HDMI interface connected to HD display. The treatments embedded in the system are as follow: (i) pre-processing such as edge detection implemented in the ARM and in the reconfigurable logic, (ii) software implementation of motion detection and face detection using either ViolaJones or LBP (Local Binary Pattern), and (iii) application layer to select processing application and to display results in a web page. One uniquely interesting feature of the proposed system is that two functions have been developed to transmit data from and to the VDMA port. With the proposed optimization, the hardware implementation of the Sobel filter takes 27 ms and 76 ms for 640x480, and 720p resolutions, respectively. Hence, with the FPGA implementation, an acceleration of 5 times is obtained which allow the processing of 37 fps and 13 fps for 640x480, and 720p resolutions, respectively.

  13. Real-time RPA assay for rapid detection and differentiation of wild-type pseudorabies and gE-deleted vaccine viruses.

    Science.gov (United States)

    Wang, Jianchang; Liu, Libing; Wang, Jinfeng; Pang, Xiaoyu; Yuan, Wanzhe

    2018-02-15

    The objective of this study was to develop a dual real-time recombinase polymerase amplification (RPA) assay using exo probes for the detection and differentiation of pseudorabies virus (PRV). Specific RPA primers and probes were designed for gB and gE genes of PRV within the conserved region of viral genome. The reaction process can be completed in 20 min at 39 °C. The dual real-time RPA assay performed in the single tube was capable of specific detecting and differentiating of the wild-type PRV and gE-deleted vaccine strains, without cross-reactions with other non-targeted pig viruses. The analytical sensitivity of the assay was 10 2 copies for gB and gE genes. The dual real-time RPA demonstrated a 100% diagnostic agreement with the real-time PCR on 4 PRV strains and 37 clinical samples. Through the linear regression analysis, the R 2 value of the real-time RPA and the real-time PCR for gB and gE was 0.983 and 0.992, respectively. The dual real-time RPA assay provides an alternative useful tool for rapid, simple, and reliable detection and differentiation of PRV, especially in remote and rural areas. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Real-time polymerase chain reaction assay for the rapid detection and characterization of chloroquine-resistant Plasmodium falciparum malaria in returned travelers.

    Science.gov (United States)

    Farcas, Gabriella A; Soeller, Rainer; Zhong, Kathleen; Zahirieh, Alireza; Kain, Kevin C

    2006-03-01

    Imported drug-resistant malaria is a growing problem in industrialized countries. Rapid and accurate diagnosis is essential to prevent malaria-associated mortality in returned travelers. However, outside of a limited number of specialized centers, the microscopic diagnosis of malaria is slow, unreliable, and provides little information about drug resistance. Molecular diagnostics have the potential to overcome these limitations. We developed and evaluated a rapid, real-time polymerase chain reaction (PCR) assay to detect Plasmodium falciparum malaria and chloroquine (CQ)-resistance determinants in returned travelers who are febrile. A real-time PCR assay based on detection of the K76T mutation in PfCRT (K76T) of P. falciparum was developed on a LightCycler platform (Roche). The performance characteristics of the real-time assay were compared with those of the nested PCR-restriction fragment-length polymorphism (RFLP) and the sequence analyses of samples obtained from 200 febrile returned travelers, who included 125 infected with P. falciparum (48 of whom were infected CQ-susceptible [K76] and 77 of whom were CQ-resistant [T76] P. falciparum), 22 infected with Plasmodium vivax, 10 infected with Plasmodium ovale, 3 infected with Plasmodium malariae malaria, and 40 infected with other febrile syndromes. All patient samples were coded, and all analyses were performed blindly. The real-time PCR assay detected multiple pfcrt haplotypes associated with CQ resistance in geographically diverse malaria isolates acquired by travelers. Compared with nested-PCR RFLP (the reference standard), the real-time assay was 100% sensitive and 96.2% specific for detection of the P. falciparum K76T mutation. This assay is rapid, sensitive, and specific for the detection and characterization of CQ-resistant P. falciparum malaria in returned travelers. This assay is automated, standardized, and suitable for routine use in clinical diagnostic laboratories.

  15. Real-time petroleum spill detection system

    International Nuclear Information System (INIS)

    Dakin, D.T.

    2001-01-01

    A real-time autonomous oil and fuel spill detection system has been developed to rapidly detect of a wide range of petroleum products floating on, or suspended in water. The system consists of an array of spill detection buoys distributed within the area to be monitored. The buoys are composed of a float and a multispectral fluorometer, which looks up through the top 5 cm of water to detect floating and suspended petroleum products. The buoys communicate to a base station computer that controls the sampling of the buoys and analyses the data from each buoy to determine if a spill has occurred. If statistically significant background petroleum levels are detected, the system raises an oil spill alarm. The system is useful because early detection of a marine oil spill allows for faster containment, thereby minimizing the contaminated area and reducing cleanup costs. This paper also provided test results for biofouling, various petroleum product detection, water turbidity and wave tolerance. The technology has been successfully demonstrated. The UV light source keeps the optic window free from biofouling, and the electronics are fully submerged so there is no risk that the unit could ignite the vapours of a potential oil spill. The system can also tolerate moderately turbid waters and can therefore be used in many rivers, harbours, water intakes and sumps. The system can detect petroleum products with an average thickness of less than 3 micrometers floating on the water surface. 3 refs., 15 figs

  16. A Rapid and Simple Real-Time PCR Assay for Detecting Foodborne Pathogenic Bacteria in Human Feces.

    Science.gov (United States)

    Hanabara, Yutaro; Ueda, Yutaka

    2016-11-22

    A rapid, simple method for detecting foodborne pathogenic bacteria in human feces is greatly needed. Here, we examined the efficacy of a method that employs a combination of a commercial PCR master mix, which is insensitive to PCR inhibitors, and a DNA extraction method which used sodium dodecyl benzene sulfonate (SDBS), and Tween 20 to counteract the inhibitory effects of SDBS on the PCR assay. This method could detect the target genes (stx1 and stx2 of enterohemorrhagic Escherichia coli, invA of Salmonella Enteritidis, tdh of Vibrio parahaemolyticus, gyrA of Campylobacter jejuni, ceuE of Campylobacter coli, SEA of Staphylococcus aureus, ces of Bacillus cereus, and cpe of Clostridium perfringens) in a fecal suspension containing 1.0 × 10 1 to 1.0 × 10 3 CFU/ml. Furthermore, the assay was neither inhibited nor influenced by individual differences among the fecal samples of 10 subjects or fecal concentration (40-160 mg/ml in the fecal suspension). When we attempted to detect the genes of pathogenic bacteria in 4 actual clinical cases, we found that this method was more sensitive than standard culture method. These results showed that this assay is a rapid, simple detection method for foodborne pathogenic bacteria in human feces.

  17. Development and implementation of real-time nucleic acid amplification for the detection of enterovirus infections in comparison to rapid culture of various clinical specimens

    NARCIS (Netherlands)

    van Doornum, G J J; Schutten, Martin; Voermans, J; Guldemeester, G J J; Niesters, H G M

    2007-01-01

    Several real-time PCR and nucleic acid sequence-based amplification (NASBA) primer pairs and a modified real-time PCR primer pair for the detection of enteroviruses were compared. The modified real-time PCR primer pair was evaluated on clinical samples in comparison with cell culture using the

  18. Multiplex, Rapid, and Sensitive Isothermal Detection of Nucleic-Acid Sequence by Endonuclease Restriction-Mediated Real-Time Multiple Cross Displacement Amplification.

    Science.gov (United States)

    Wang, Yi; Wang, Yan; Zhang, Lu; Liu, Dongxin; Luo, Lijuan; Li, Hua; Cao, Xiaolong; Liu, Kai; Xu, Jianguo; Ye, Changyun

    2016-01-01

    We have devised a novel isothermal amplification technology, termed endonuclease restriction-mediated real-time multiple cross displacement amplification (ET-MCDA), which facilitated multiplex, rapid, specific and sensitive detection of nucleic-acid sequences at a constant temperature. The ET-MCDA integrated multiple cross displacement amplification strategy, restriction endonuclease cleavage and real-time fluorescence detection technique. In the ET-MCDA system, the functional cross primer E-CP1 or E-CP2 was constructed by adding a short sequence at the 5' end of CP1 or CP2, respectively, and the new E-CP1 or E-CP2 primer was labeled at the 5' end with a fluorophore and in the middle with a dark quencher. The restriction endonuclease Nb.BsrDI specifically recognized the short sequence and digested the newly synthesized double-stranded terminal sequences (5' end short sequences and their complementary sequences), which released the quenching, resulting on a gain of fluorescence signal. Thus, the ET-MCDA allowed real-time detection of single or multiple targets in only a single reaction, and the positive results were observed in as short as 12 min, detecting down to 3.125 fg of genomic DNA per tube. Moreover, the analytical specificity and the practical application of the ET-MCDA were also successfully evaluated in this study. Here, we provided the details on the novel ET-MCDA technique and expounded the basic ET-MCDA amplification mechanism.

  19. Rapid detection of Enterovirus and Coxsackievirus A10 by a TaqMan based duplex one-step real time RT-PCR assay.

    Science.gov (United States)

    Chen, Jingfang; Zhang, Rusheng; Ou, Xinhua; Yao, Dong; Huang, Zheng; Li, Linzhi; Sun, Biancheng

    2017-06-01

    A TaqMan based duplex one-step real time RT-PCR (rRT-PCR) assay was developed for the rapid detection of Coxsackievirus A10 (CV-A10) and other enterovirus (EVs) in clinical samples. The assay was fully evaluated and found to be specific and sensitive. When applied in 115 clinical samples, a 100% diagnostic sensitivity in CV-A10 detection and 97.4% diagnostic sensitivity in other EVs were found. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Rapid detection of Listeria monocytogenes in raw milk and soft cheese by a redox potential measurement based method combined with real-time PCR.

    Science.gov (United States)

    Erdősi, Orsolya; Szakmár, Katalin; Reichart, Olivér; Szili, Zsuzsanna; László, Noémi; Székely Körmöczy, Péter; Laczay, Péter

    2014-09-01

    The incidence of outbreaks of foodborne listeriosis has indicated the need for a reliable and rapid detection of the microbe in different foodstuffs. A method combining redox potential measurement and real-time polymerase chain reaction (PCR) was developed to detect Listeria monocytogenes in artificially contaminated raw milk and soft cheese. Food samples of 25 g or 25 ml were homogenised in 225 ml of Listeria Enrichment Broth (LEB) with Oxford supplement, and the redox potential measurement technique was applied. For Listeria species the measuring time was maximum 34 h. The absence of L. monocytogenes could reliably be proven by the redox potential measurement method, but Listeria innocua and Bacillus subtilis could not be differentiated from L. monocytogenes on the basis of the redox curves. The presence of L. monocytogenes had to be confirmed by real-time PCR. The combination of these two methods proved to detect < 10 cfu/g of L. monocytogenes in a cost- and time-effective manner. This method can potentially be used as an alternative to the standard nutrient method for the rapid detection of L. monocytogenes in food.

  1. Rapid detection of Opisthorchis viverrini and Strongyloides stercoralis in human fecal samples using a duplex real-time PCR and melting curve analysis.

    Science.gov (United States)

    Janwan, Penchom; Intapan, Pewpan M; Thanchomnang, Tongjit; Lulitanond, Viraphong; Anamnart, Witthaya; Maleewong, Wanchai

    2011-12-01

    Human opisthorchiasis caused by the liver fluke Opisthorchis viverrini is an endemic disease in Southeast Asian countries including the Lao People's Democratic Republic, Cambodia, Vietnam, and Thailand. Infection with the soil-transmitted roundworm Strongyloides stercoralis is an important problem worldwide. In some areas, both parasitic infections are reported as co-infections. A duplex real-time fluorescence resonance energy transfer (FRET) PCR merged with melting curve analysis was developed for the rapid detection of O. viverrini and S. stercoralis in human fecal samples. Duplex real-time FRET PCR is based on fluorescence melting curve analysis of a hybrid of amplicons generated from two genera of DNA elements: the 162 bp pOV-A6 DNA sequence specific to O. viverrini and the 244 bp 18S rRNA sequence specific to S. stercoralis, and two pairs of specific fluorophore-labeled probes. Both O. viverrini and S. stercoralis can be differentially detected in infected human fecal samples by this process through their different fluorescence channels and melting temperatures. Detection limit of the method was as little as two O. viverrini eggs and four S. stercoralis larvae in 100 mg of fecal sample. The assay could distinguish the DNA of both parasites from the DNA of negative fecal samples and fecal samples with other parasite materials, as well as from the DNA of human leukocytes and other control parasites. The technique showed 100% sensitivity and specificity. The introduced duplex real-time FRET PCR can reduce labor time and reagent costs and is not prone to carry over contamination. The method is important for simultaneous detection especially in areas where both parasites overlap incidence and is useful as the screening tool in the returning travelers and immigrants to industrialized countries where number of samples in the diagnostic units will become increasing.

  2. Multiplex, rapid and sensitive isothermal detection of nucleic-acid sequence by endonuclease restriction-mediated real-time multiple cross displacement amplification

    Directory of Open Access Journals (Sweden)

    Yi eWang

    2016-05-01

    Full Text Available We have devised a novel isothermal amplification technology, termed endonuclease restriction-mediated real-time multiple cross displacement amplification (ET-MCDA, which facilitated multiplex, rapid, specific and sensitive detection of nucleic-acid sequences at a constant temperature. The ET-MCDA integrated multiple cross displacement amplification strategy, restriction endonuclease cleavage and real-time fluorescence detection technique. In the ET-MCDA system, the functional cross primer E-CP1 or E-CP2 was constructed by adding a short sequence at the 5’ end of CP1 or CP2, respectively, and the new E-CP1 or E-CP2 primer was labelled at the 5’ end with a fluorophore and in the middle with a dark quencher. The restriction endonuclease Nb.BsrDI specifically recognized the short sequence and digested the newly synthesized double-stranded terminal sequences (5’ end short sequences and their complementary sequences, which released the quenching, resulting on a gain of fluorescence signal. Thus, the ET-MCDA allowed real-time detection of single or multiple targets in only a single reaction, and the positive results were observed in as short as 12 minutes, detecting down to 3.125 fg of genomic DNA per tube. Moreover, the analytical specificity and the practical application of the ET-MCDA were also successfully evaluated in this study. Here we provided the details on the novel ET-MCDA technique and expounded the basic ET-MCDA amplification mechanism.

  3. Performance of the new automated Abbott RealTime MTB assay for rapid detection of Mycobacterium tuberculosis complex in respiratory specimens.

    Science.gov (United States)

    Chen, J H K; She, K K K; Kwong, T-C; Wong, O-Y; Siu, G K H; Leung, C-C; Chang, K-C; Tam, C-M; Ho, P-L; Cheng, V C C; Yuen, K-Y; Yam, W-C

    2015-09-01

    The automated high-throughput Abbott RealTime MTB real-time PCR assay has been recently launched for Mycobacterium tuberculosis complex (MTBC) clinical diagnosis. This study would like to evaluate its performance. We first compared its diagnostic performance with the Roche Cobas TaqMan MTB assay on 214 clinical respiratory specimens. Prospective analysis of a total 520 specimens was then performed to further evaluate the Abbott assay. The Abbott assay showed a lower limit of detection at 22.5 AFB/ml, which was more sensitive than the Cobas assay (167.5 AFB/ml). The two assays demonstrated a significant difference in diagnostic performance (McNemar's test; P = 0.0034), in which the Abbott assay presented significantly higher area under curve (AUC) than the Cobas assay (1.000 vs 0.880; P = 0.0002). The Abbott assay demonstrated extremely low PCR inhibition on clinical respiratory specimens. The automated Abbott assay required only very short manual handling time (0.5 h), which could help to improve the laboratory management. In the prospective analysis, the overall estimates for sensitivity and specificity of the Abbott assay were both 100 % among smear-positive specimens, whereas the smear-negative specimens were 96.7 and 96.1 %, respectively. No cross-reactivity with non-tuberculosis mycobacterial species was observed. The superiority in sensitivity of the Abbott assay for detecting MTBC in smear-negative specimens could further minimize the risk in MTBC false-negative detection. The new Abbott RealTime MTB assay has good diagnostic performance which can be a useful diagnostic tool for rapid MTBC detection in clinical laboratories.

  4. Rapid detection of most frequent Slovenian germ-line mutations in BRCA1 gene using real-time PCR and melting curve analysis

    International Nuclear Information System (INIS)

    Novakovic, S.; Stegel, V.

    2005-01-01

    Background. Detection of inherited mutations in cancer susceptibility genes is of great importance in some types of cancers including the colorectal cancer (mutations of APC gene in familial adenomatous polyposis - FAP, mutations in mismatch repair genes in hereditary nonpolyposis colorectal cancer - HNPCC), malignant melanoma (mutations in CDKN2A and CDK4 genes) and breast cancer (mutations in BRCA1 and BRCA2 genes). Methods. This article presents the technical data for the detection of five mutations in BRCA1 gene in breast cancer patients and their relatives. The mutations - 1806C>T, 300T>G, 300T>A, 310G>A, 5382insC - were determined by the real-time PCR and the melting curve analysis. Results and conclusion. In comparison to direct sequencing, this method proved to be sensitive and rapid enough for the routine daily determination of mutations in DNA isolated from the peripheral blood. (author)

  5. Rapid and Sensitive Detection of Yersinia pestis Using Amplification of Plague Diagnostic Bacteriophages Monitored by Real-Time PCR

    Science.gov (United States)

    2010-06-01

    2004) Development of a bacteriophage phage replication assay for diagnosis of pulmonary tuberculosis . J Clin Microbiol 42: 2115–2120. 30. Abshire TG...Mycobacterium tuberculosis in Mexico . Clin Microbiol 39: 3883–3888. 38. Schofield DA, Westwater C (2009) Phage-mediated bioluminescent detection of

  6. Rapid and Specific Detection of Acidovorax avenae subsp. citrulli Using SYBR Green-Based Real-Time PCR Amplification of the YD-Repeat Protein Gene.

    Science.gov (United States)

    Cho, Min Seok; Park, Duck Hwan; Ahn, Tae-Young; Park, Dong Suk

    2015-09-01

    The aim of this study was to develop a SYBR Green-based real-time PCR assay for the rapid, specific, and sensitive detection of Acidovorax avenae subsp. citrulli, which causes bacterial fruit blotch (BFB), a serious disease of cucurbit plants. The molecular and serological methods currently available for the detection of this pathogen are insufficiently sensitive and specific. Thus, a novel SYBR Green-based real-time PCR assay targeting the YD-repeat protein gene of A. avenae subsp. citrulli was developed. The specificity of the primer set was evaluated using DNA purified from 6 isolates of A. avenae subsp. citrulli, 7 other Acidovorax species, and 22 of non-targeted strains, including pathogens and non-pathogens. The AC158F/R primer set amplified a single band of the expected size from genomic DNA obtained from the A. avenae subsp. citrulli strains but not from the genomic DNA of other Acidovorax species, including that of other bacterial genera. Using this assay, it was possible to detect at least one genomeequivalents of the cloned amplified target DNA using 5 × 10(0) fg/μl of purified genomic DNA per reaction or using a calibrated cell suspension, with 6.5 colony-forming units per reaction being employed. In addition, this assay is a highly sensitive and reliable method for identifying and quantifying the target pathogen in infected samples that does not require DNA extraction. Therefore, we suggest that this approach is suitable for the rapid and efficient diagnosis of A. avenae subsp. citrulli contaminations of seed lots and plants.

  7. Multiplex real-time quantitative PCR, microscopy and rapid diagnostic immuno-chromatographic tests for the detection of Plasmodium spp: performance, limit of detection analysis and quality assurance

    Directory of Open Access Journals (Sweden)

    Ralevski Filip

    2009-12-01

    Full Text Available Abstract Background Accurate laboratory diagnosis of malaria species in returning travelers is paramount in the treatment of this potentially fatal infectious disease. Materials and methods A total of 466 blood specimens from returning travelers to Africa, Asia, and South/Central America with suspected malaria infection were collected between 2007 and 2009 at the reference public health laboratory. These specimens were assessed by reference microscopy, multipex real-time quantitative polymerase chain reaction (QPCR, and two rapid diagnostic immuno-chromatographic tests (ICT in a blinded manner. Key clinical laboratory parameters such as limit of detection (LOD analysis on clinical specimens by parasite stage, inter-reader variability of ICTs, staffing implications, quality assurance and cost analysis were evaluated. Results QPCR is the most analytically sensitive method (sensitivity 99.41%, followed by CARESTART (sensitivity 88.24%, and BINAXNOW (sensitivity 86.47% for the diagnosis of malaria in returning travelers when compared to reference microscopy. However, microscopy was unable to specifically identify Plasmodia spp. in 18 out of 170 positive samples by QPCR. Moreover, the 17 samples that were negative by microscopy and positive by QPCR were also positive by ICTs. Quality assurance was achieved for QPCR by exchanging a blinded proficiency panel with another reference laboratory. The Kappa value of inter-reader variability among three readers for BINAXNOW and CARESTART was calculated to be 0.872 and 0.898 respectively. Serial dilution studies demonstrated that the QPCR cycle threshold correlates linearly with parasitemia (R2 = 0.9746 in a clinically relevant dynamic range and retains a LOD of 11 rDNA copies/μl for P. falciparum, which was several log lower than reference microscopy and ICTs. LOD for QPCR is affected not only by parasitemia but the parasite stage distribution of each clinical specimen. QPCR was approximately 6-fold more

  8. A locked nucleic acid (LNA-based real-time PCR assay for the rapid detection of multiple bacterial antibiotic resistance genes directly from positive blood culture.

    Directory of Open Access Journals (Sweden)

    Lingxiang Zhu

    Full Text Available Bacterial strains resistant to various antibiotic drugs are frequently encountered in clinical infections, and the rapid identification of drug-resistant strains is highly essential for clinical treatment. We developed a locked nucleic acid (LNA-based quantitative real-time PCR (LNA-qPCR method for the rapid detection of 13 antibiotic resistance genes and successfully used it to distinguish drug-resistant bacterial strains from positive blood culture samples. A sequence-specific primer-probe set was designed, and the specificity of the assays was assessed using 27 ATCC bacterial strains and 77 negative blood culture samples. No cross-reaction was identified among bacterial strains and in negative samples, indicating 100% specificity. The sensitivity of the assays was determined by spiking each bacterial strain into negative blood samples, and the detection limit was 1-10 colony forming units (CFU per reaction. The LNA-qPCR assays were first applied to 72 clinical bacterial isolates for the identification of known drug resistance genes, and the results were verified by the direct sequencing of PCR products. Finally, the LNA-qPCR assays were used for the detection in 47 positive blood culture samples, 19 of which (40.4% were positive for antibiotic resistance genes, showing 91.5% consistency with phenotypic susceptibility results. In conclusion, LNA-qPCR is a reliable method for the rapid detection of bacterial antibiotic resistance genes and can be used as a supplement to phenotypic susceptibility testing for the early detection of antimicrobial resistance to allow the selection of appropriate antimicrobial treatment and to prevent the spread of resistant isolates.

  9. Rapid and sensitive detection of Pseudomonas aeruginosa in chlorinated water and aerosols targeting gyrB gene using real-time PCR.

    Science.gov (United States)

    Lee, C S; Wetzel, K; Buckley, T; Wozniak, D; Lee, J

    2011-10-01

    For the rapid detection of Pseudomonas aeruginosa from chlorinated water and aerosols, gyrB gene-based real-time PCR assay was developed and investigated. Two novel primer sets (pa722F/746MGB/899R and pa722F/746MGB/788R) were designed using the most updated 611 Pseudomonas and 748 other bacterial gyrB genes for achieving high specificity. Their specificity showed 100% accuracy when tested with various strains including clinical isolates from cystic fibrosis patients. The assay was tested with Ps. aeruginosa-containing chlorinated water and aerosols to simulate the waterborne and airborne transmission routes (detection limit 3·3 × 10² CFU per PCR-2·3 × 10³ CFU per PCR). No chlorine interference in real-time PCR was observed at drinking water level (c. 1 mg l⁻¹), but high level of chorine (12 mg l⁻¹) interfered the assay, and thus neutralization was needed. Pseudomonas aeruginosa in aerosol was successfully detected after capturing with gelatin filters with minimum 2 min of sampling time when the initial concentration of 10⁴ CFU ml⁻¹ bacteria existed in the nebulizer. A highly specific and rapid assay (2-3 h) was developed by targeting gyrB gene for the detection of Ps. aeruginosa in chlorinated water and aerosols, combined with optimized sample collection methods and sample processing, so the direct DNA extraction from either water or aerosol was possible while achieving the desired sensitivity of the method.   The new assay can provide timely and accurate risk assessment to prevent Ps. aeruginosa exposure from water and aerosol, resulting in reduced disease burden, especially among immune-compromised and susceptible individuals. This approach can be easily utilized as a platform technology for the detection of other types of micro-organisms, especially for those that are transmitted via water and aerosol routes, such as Legionella pneumophila. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  10. Rapid detection of health-care-associated bloodstream infection in critical care using multipathogen real-time polymerase chain reaction technology: a diagnostic accuracy study and systematic review.

    Science.gov (United States)

    Warhurst, Geoffrey; Dunn, Graham; Chadwick, Paul; Blackwood, Bronagh; McAuley, Daniel; Perkins, Gavin D; McMullan, Ronan; Gates, Simon; Bentley, Andrew; Young, Duncan; Carlson, Gordon L; Dark, Paul

    2015-05-01

    antibiotic exposure. SeptiFast real-time PCR, when compared with culture-proven bloodstream infection at species/genus level, had better specificity (85.8%, 95% CI 83.3% to 88.1%) than sensitivity (50%, 95% CI 39.1% to 60.8%). When compared with pooled diagnostic metrics derived from our systematic review, our clinical study revealed lower test accuracy of SeptiFast real-time PCR, mainly as a result of low diagnostic sensitivity. There was a low prevalence of BC-proven pathogens in these patients (9.2%, 95% CI 7.4% to 11.2%) such that the post-test probabilities of both a positive (26.3%, 95% CI 19.8% to 33.7%) and a negative SeptiFast test (5.6%, 95% CI 4.1% to 7.4%) indicate the potential limitations of this technology in the diagnosis of bloodstream infection. However, latent class analysis indicates that BC has a low sensitivity, questioning its relevance as a reference test in this setting. Using this analysis approach, the sensitivity of the SeptiFast test was low but also appeared significantly better than BC. Blood samples identified as positive by either culture or SeptiFast real-time PCR were associated with a high probability (> 95%) of infection, indicating higher diagnostic rule-in utility than was apparent using conventional analyses of diagnostic accuracy. SeptiFast real-time PCR on blood samples may have rapid rule-in utility for the diagnosis of health-care-associated bloodstream infection but the lack of sensitivity is a significant limiting factor. Innovations aimed at improved diagnostic sensitivity of real-time PCR in this setting are urgently required. Future work recommendations include technology developments to improve the efficiency of pathogen DNA extraction and the capacity to detect a much broader range of pathogens and drug resistance genes and the application of new statistical approaches able to more reliably assess test performance in situation where the reference standard (e.g. blood culture in the setting of high antimicrobial use) is

  11. Development of a taqman-based real-time PCR assay for the rapid and specific detection of novel duck- origin goose parvovirus.

    Science.gov (United States)

    Wang, Jianchang; Wang, Jinfeng; Cui, Yuan; Nan, Huizhu; Yuan, Wanzhe

    2017-08-01

    A real-time PCR assay was developed for specific detection of novel duck-origin goose parvovirus (N-GPV), the etiological agent of duck beak atrophy and dwarfism syndrome (BADS). The detection limit of the assay was 10 2 copies. The assay was useful in the prevention and control of BADS. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Near Real Time Ship Detection Experiments

    Science.gov (United States)

    Brusch, S.; Lehner, S.; Schwarz, E.; Fritz, T.

    2010-04-01

    A new Near Real Time (NRT) ship detection processor SAINT (SAR AIS Integrated Toolbox) was developed in the framework of the ESA project MARISS. Data are received at DLRs ground segment DLR-BN (Neustrelitz, Germany). Results of the ship detection are available on ftp server within 30 min after the acquisition started. The detectability of ships on Synthetic Aperture Radar (SAR) ERS-2, ENVISAT ASAR and TerraSAR-X (TS-X) images is validated by coastal (live) AIS and space AIS. The monitoring areas chosen for surveillance are the North-, Baltic Sea, and Cape Town. The detectability in respect to environmental parameters like wind field, sea state, currents and changing coastlines due to tidal effects is investigated. In the South Atlantic a tracking experiment of the German research vessel Polarstern has been performed. Issues of piracy in particular in respect to ships hijacked at the Somali coast are discussed. Some examples using high resolution images from TerraSAR-X are given.

  13. Development of a Taqman real-time PCR assay for rapid detection and quantification of Vibrio tapetis in extrapallial fluids of clams

    Directory of Open Access Journals (Sweden)

    Adeline Bidault

    2015-12-01

    Full Text Available The Gram-negative bacterium Vibrio tapetis is known as the causative agent of Brown Ring Disease (BRD in the Manila clam Venerupis (=Ruditapes philippinarum. This bivalve is the second most important species produced in aquaculture and has a high commercial value. In spite of the development of several molecular methods, no survey has been yet achieved to rapidly quantify the bacterium in the clam. In this study, we developed a Taqman real-time PCR assay targeting virB4 gene for accurate and quantitative identification of V. tapetis strains pathogenic to clams. Sensitivity and reproducibility of the method were assessed using either filtered sea water or extrapallial fluids of clam injected with the CECT4600T V. tapetis strain. Quantification curves of V. tapetis strain seeded in filtered seawater (FSW or extrapallial fluids (EF samples were equivalent showing reliable qPCR efficacies. With this protocol, we were able to specifically detect V. tapetis strains down to 1.125 101 bacteria per mL of EF or FSW, taking into account the dilution factor used for appropriate template DNA preparation. This qPCR assay allowed us to monitor V. tapetis load both experimentally or naturally infected Manila clams. This technique will be particularly useful for monitoring the kinetics of massive infections by V. tapetis and for designing appropriate control measures for aquaculture purposes.

  14. Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR

    OpenAIRE

    Aebischer, Andrea; Wernike, Kerstin; Hoffmann, Bernd; Beer, Martin

    2014-01-01

    Over the past few years, there has been an increasing demand for rapid and simple diagnostic tools that can be applied outside centralized laboratories by using transportable devices. In veterinary medicine, such mobile test systems would circumvent barriers associated with the transportation of samples and significantly reduce the time to diagnose important infectious animal diseases. Among a wide range of available technologies, high-speed real-time reverse transcriptase quantitative PCR (R...

  15. Rapid genome detection of Schmallenberg virus and bovine viral diarrhea virus by use of isothermal amplification methods and high-speed real-time reverse transcriptase PCR.

    Science.gov (United States)

    Aebischer, Andrea; Wernike, Kerstin; Hoffmann, Bernd; Beer, Martin

    2014-06-01

    Over the past few years, there has been an increasing demand for rapid and simple diagnostic tools that can be applied outside centralized laboratories by using transportable devices. In veterinary medicine, such mobile test systems would circumvent barriers associated with the transportation of samples and significantly reduce the time to diagnose important infectious animal diseases. Among a wide range of available technologies, high-speed real-time reverse transcriptase quantitative PCR (RT-qPCR) and the two isothermal amplification techniques loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) represent three promising candidates for integration into mobile pen-side tests. The aim of this study was to investigate the performance of these amplification strategies and to evaluate their suitability for field application. In order to enable a valid comparison, novel pathogen-specific assays have been developed for the detection of Schmallenberg virus and bovine viral diarrhea virus. The newly developed assays were evaluated in comparison with established standard RT-qPCR using samples from experimentally or field-infected animals. Even though all assays allowed detection of the target virus in less than 30 min, major differences were revealed concerning sensitivity, specificity, robustness, testing time, and complexity of assay design. These findings indicated that the success of an assay will depend on the integrated amplification technology. Therefore, the application-specific pros and cons of each method that were identified during this study provide very valuable insights for future development and optimization of pen-side tests. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  16. Real time detecting system for turning force

    Energy Technology Data Exchange (ETDEWEB)

    Xiaobin, Yue [China Academy of Engineering Physics, Mianyang (China). Inst. of Machinery Manufacturing Technology

    2001-07-01

    How to get the real-time value of forces dropped on the tool in the course of processing by piezoelectric sensors is introduced. First, the analog signals of the cutting force were achieved by these sensors, amplified and transferred into digital signals by A/D transferring card. Then real-time software reads the information, put it into its own coordinate, drew the curve of forces, displayed it on the screen by the real time and saved it for the technicians to analyze the situation of the tool. So the cutting parameter can be optimized to improve surface quality of the pieces.

  17. Linear-after-the-exponential polymerase chain reaction and allied technologies. Real-time detection strategies for rapid, reliable diagnosis from single cells.

    Science.gov (United States)

    Pierce, Kenneth E; Wangh, Lawrence J

    2007-01-01

    Accurate detection of gene sequences in single cells is the ultimate challenge to polymerase chain reaction (PCR) sensitivity. Unfortunately, commonly used conventional and real-time PCR techniques are often too unreliable at that level to provide the accuracy needed for clinical diagnosis. Here we provide details of linear-after-the-exponential-PCR (LATE-PCR), a method similar to asymmetric PCR in the use of primers at different concentrations, but with novel design criteria to ensure high efficiency and specificity. Compared with conventional PCR, LATE-PCR increases the signal strength and allele discrimination capability of oligonucleotide probes such as molecular beacons and reduces variability among replicate samples. The analysis of real-time kinetics of LATE-PCR signals provides a means for improving the accuracy of single cell genetic diagnosis.

  18. Cross-platform evaluation of commercial real-time SYBR green RT-PCR kits for sensitive and rapid detection of European bat Lyssavirus type 1.

    Science.gov (United States)

    Picard-Meyer, Evelyne; Peytavin de Garam, Carine; Schereffer, Jean Luc; Marchal, Clotilde; Robardet, Emmanuelle; Cliquet, Florence

    2015-01-01

    This study evaluates the performance of five two-step SYBR Green RT-qPCR kits and five one-step SYBR Green qRT-PCR kits using real-time PCR assays. Two real-time thermocyclers showing different throughput capacities were used. The analysed performance evaluation criteria included the generation of standard curve, reaction efficiency, analytical sensitivity, intra- and interassay repeatability as well as the costs and the practicability of kits, and thermocycling times. We found that the optimised one-step PCR assays had a higher detection sensitivity than the optimised two-step assays regardless of the machine used, while no difference was detected in reaction efficiency, R (2) values, and intra- and interreproducibility between the two methods. The limit of detection at the 95% confidence level varied between 15 to 981 copies/µL and 41 to 171 for one-step kits and two-step kits, respectively. Of the ten kits tested, the most efficient kit was the Quantitect SYBR Green qRT-PCR with a limit of detection at 95% of confidence of 20 and 22 copies/µL on the thermocyclers Rotor gene Q MDx and MX3005P, respectively. The study demonstrated the pivotal influence of the thermocycler on PCR performance for the detection of rabies RNA, as well as that of the master mixes.

  19. Cross-Platform Evaluation of Commercial Real-Time SYBR Green RT-PCR Kits for Sensitive and Rapid Detection of European Bat Lyssavirus Type 1

    Directory of Open Access Journals (Sweden)

    Evelyne Picard-Meyer

    2015-01-01

    Full Text Available This study evaluates the performance of five two-step SYBR Green RT-qPCR kits and five one-step SYBR Green qRT-PCR kits using real-time PCR assays. Two real-time thermocyclers showing different throughput capacities were used. The analysed performance evaluation criteria included the generation of standard curve, reaction efficiency, analytical sensitivity, intra- and interassay repeatability as well as the costs and the practicability of kits, and thermocycling times. We found that the optimised one-step PCR assays had a higher detection sensitivity than the optimised two-step assays regardless of the machine used, while no difference was detected in reaction efficiency, R2 values, and intra- and interreproducibility between the two methods. The limit of detection at the 95% confidence level varied between 15 to 981 copies/µL and 41 to 171 for one-step kits and two-step kits, respectively. Of the ten kits tested, the most efficient kit was the Quantitect SYBR Green qRT-PCR with a limit of detection at 95% of confidence of 20 and 22 copies/µL on the thermocyclers Rotor gene Q MDx and MX3005P, respectively. The study demonstrated the pivotal influence of the thermocycler on PCR performance for the detection of rabies RNA, as well as that of the master mixes.

  20. Polymeric LabChip real-time PCR as a point-of-care-potential diagnostic tool for rapid detection of influenza A/H1N1 virus in human clinical specimens.

    Directory of Open Access Journals (Sweden)

    Hyun-Ok Song

    Full Text Available It is clinically important to be able to detect influenza A/H1N1 virus using a fast, portable, and accurate system that has high specificity and sensitivity. To achieve this goal, it is necessary to develop a highly specific primer set that recognizes only influenza A viral genes and a rapid real-time PCR system that can detect even a single copy of the viral gene. In this study, we developed and validated a novel fluidic chip-type real-time PCR (LabChip real-time PCR system that is sensitive and specific for the detection of influenza A/H1N1, including the pandemic influenza strain A/H1N1 of 2009. This LabChip real-time PCR system has several remarkable features: (1 It allows rapid quantitative analysis, requiring only 15 min to perform 30 cycles of real-time PCR. (2 It is portable, with a weight of only 5.5 kg. (3 The reaction cost is low, since it uses disposable plastic chips. (4 Its high efficiency is equivalent to that of commercially available tube-type real-time PCR systems. The developed disposable LabChip is an economic, heat-transferable, light-transparent, and easy-to-fabricate polymeric chip compared to conventional silicon- or glass-based labchip. In addition, our LabChip has large surface-to-volume ratios in micro channels that are required for overcoming time consumed for temperature control during real-time PCR. The efficiency of the LabChip real-time PCR system was confirmed using novel primer sets specifically targeted to the hemagglutinin (HA gene of influenza A/H1N1 and clinical specimens. Eighty-five human clinical swab samples were tested using the LabChip real-time PCR. The results demonstrated 100% sensitivity and specificity, showing 72 positive and 13 negative cases. These results were identical to those from a tube-type real-time PCR system. This indicates that the novel LabChip real-time PCR may be an ultra-fast, quantitative, point-of-care-potential diagnostic tool for influenza A/H1N1 with a high sensitivity and

  1. Approaching near real-time biosensing: microfluidic microsphere based biosensor for real-time analyte detection.

    Science.gov (United States)

    Cohen, Noa; Sabhachandani, Pooja; Golberg, Alexander; Konry, Tania

    2015-04-15

    In this study we describe a simple lab-on-a-chip (LOC) biosensor approach utilizing well mixed microfluidic device and a microsphere-based assay capable of performing near real-time diagnostics of clinically relevant analytes such cytokines and antibodies. We were able to overcome the adsorption kinetics reaction rate-limiting mechanism, which is diffusion-controlled in standard immunoassays, by introducing the microsphere-based assay into well-mixed yet simple microfluidic device with turbulent flow profiles in the reaction regions. The integrated microsphere-based LOC device performs dynamic detection of the analyte in minimal amount of biological specimen by continuously sampling micro-liter volumes of sample per minute to detect dynamic changes in target analyte concentration. Furthermore we developed a mathematical model for the well-mixed reaction to describe the near real time detection mechanism observed in the developed LOC method. To demonstrate the specificity and sensitivity of the developed real time monitoring LOC approach, we applied the device for clinically relevant analytes: Tumor Necrosis Factor (TNF)-α cytokine and its clinically used inhibitor, anti-TNF-α antibody. Based on the reported results herein, the developed LOC device provides continuous sensitive and specific near real-time monitoring method for analytes such as cytokines and antibodies, reduces reagent volumes by nearly three orders of magnitude as well as eliminates the washing steps required by standard immunoassays. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Rapid and Quantitative Detection of Leifsonia xyli subsp. xyli in Sugarcane Stalk Juice Using a Real-Time Fluorescent (TaqMan PCR Assay

    Directory of Open Access Journals (Sweden)

    Hua-Ying Fu

    2016-01-01

    Full Text Available Ratoon stunting disease (RSD of sugarcane, one of the most important diseases seriously affecting the productivity of sugarcane crops, was caused by the bacterial agent Leifsonia xyli subsp. xyli (Lxx. A TaqMan probe-based real-time quantitative polymerase chain reaction (qPCR assay was established in this study for the quantification of Lxx detection in sugarcane stalk juice. A pair of PCR primers (Pat1-QF/Pat1-QR and a fluorogenic probe (Pat1-QP targeting the Part1 gene of Lxx were used for the qPCR assay. The assay had a detection limit of 100 copies of plasmid DNA and 100 fg of Lxx genomic DNA, which was 100-fold more sensitive than the conventional PCR. Fifty (28.7% of 174 stalk juice samples from two field trials were tested to be positive by qPCR assay, whereas, by conventional PCR, only 12.1% (21/174 were tested to be positive with a published primer pair CxxITSf#5/CxxITSr#5 and 15.5% (27/174 were tested to be positive with a newly designed primer pair Pat1-F2/Pat1-R2. The new qPCR assay can be used as an alternative to current diagnostic methods for Lxx, especially when dealing with certificating a large number of healthy cane seedlings and determining disease incidence accurately in commercial fields.

  3. Detection of dopamine neurotransmission in 'real time'

    Directory of Open Access Journals (Sweden)

    Rajendra D Badgaiyan

    2013-07-01

    Full Text Available Current imaging techniques have limited ability to detect neurotransmitters released during brain processing. It is a critical limitation because neurotransmitters have significant control over the brain activity. In this context, recent development of single-scan dynamic molecular imaging technique is important because it allows detection, mapping, and measurement of dopamine released in the brain during task performance. The technique exploits the competition between endogenously released dopamine and its receptor ligand for occupancy of receptor sites. Dopamine released during task performance is detected by dynamically measuring concentration of intravenously injected radiolabeled ligand using a positron emission tomography camera. Based on the ligand concentration, values of receptor kinetic parameters are estimated. These estimates allow detection of dopamine released in the human brain during task performance.

  4. The use of real-time polymerase chain reaction for rapid diagnosis of skeletal tuberculosis.

    Science.gov (United States)

    Kobayashi, Naomi; Fraser, Thomas G; Bauer, Thomas W; Joyce, Michael J; Hall, Gerri S; Tuohy, Marion J; Procop, Gary W

    2006-07-01

    We identified Mycobacterium tuberculosis DNA using real-time polymerase chain reaction on a specimen from an osteolytic lesion of a femoral condyle, in which the frozen section demonstrated granulomas. The process was much more rapid than is possible with culture. The rapid detection of M tuberculosis and the concomitant exclusion of granulomatous disease caused by nontuberculous mycobacteria or systemic fungi are necessary to appropriately treat skeletal tuberculosis. The detection and identification of M tuberculosis by culture may require several weeks using traditional methods. The real-time polymerase chain reaction method used has been shown to be rapid and reliable, and is able to detect and differentiate both tuberculous and nontuberculous mycobacteria. Real-time polymerase chain reaction may become a diagnostic standard for the evaluation of clinical specimens for the presence of mycobacteria; this case demonstrates the potential utility of this assay for the rapid diagnosis of skeletal tuberculosis.

  5. Development of a multiplex real-time PCR for the rapid detection of the predominant beta-lactamase genes CTX-M, SHV, TEM and CIT-type AmpCs in Enterobacteriaceae.

    Directory of Open Access Journals (Sweden)

    Nicole Roschanski

    Full Text Available Beta-lactamase resistant bacteria and especially ESBL producing Enterobacteriaceae are an increasing problem worldwide. For this reason a major interest in efficient and reliable methods for rapid screening of high sample numbers is recognizable. Therefore, a multiplex real-time PCR was developed to detect the predominant class A beta-lactamase genes blaCTX-M, blaSHV, blaTEM and CIT-type AmpCs in a one-step reaction. A set of 114 Enterobacteriaceae containing previously identified resistance gene subtypes and in addition 20 undefined animal and environmental isolates were used for the validation of this assay. To confirm the accessibility in variable settings, the real-time runs were performed analogous in two different laboratories using different real-time cyclers. The obtained results showed complete accordance between the real-time data and the predetermined genotypes. Even if sequence analyses are further necessary for a comprehensive characterization, this method was proofed to be reliable for rapid screening of high sample numbers and therefore could be an important tool for e. g. epidemiological purposes or support infection control measures.

  6. Real-Time PCR for Universal Phytoplasma Detection and Quantification

    DEFF Research Database (Denmark)

    Christensen, Nynne Meyn; Nyskjold, Henriette; Nicolaisen, Mogens

    2013-01-01

    Currently, the most efficient detection and precise quantification of phytoplasmas is by real-time PCR. Compared to nested PCR, this method is less sensitive to contamination and is less work intensive. Therefore, a universal real-time PCR method will be valuable in screening programs and in other...

  7. Rapid detection and subtyping of European swine influenza viruses in porcine clinical samples by haemagglutinin- and neuraminidase-specific tetra- and triplex real-time RT-PCRs.

    Science.gov (United States)

    Henritzi, Dinah; Zhao, Na; Starick, Elke; Simon, Gaelle; Krog, Jesper S; Larsen, Lars Erik; Reid, Scott M; Brown, Ian H; Chiapponi, Chiara; Foni, Emanuela; Wacheck, Silke; Schmid, Peter; Beer, Martin; Hoffmann, Bernd; Harder, Timm C

    2016-11-01

    A diversifying pool of mammalian-adapted influenza A viruses (IAV) with largely unknown zoonotic potential is maintained in domestic swine populations worldwide. The most recent human influenza pandemic in 2009 was caused by a virus with genes originating from IAV isolated from swine. Swine influenza viruses (SIV) are widespread in European domestic pig populations and evolve dynamically. Knowledge regarding occurrence, spread and evolution of potentially zoonotic SIV in Europe is poorly understood. Efficient SIV surveillance programmes depend on sensitive and specific diagnostic methods which allow for cost-effective large-scale analysis. New SIV haemagglutinin (HA) and neuraminidase (NA) subtype- and lineage-specific multiplex real-time RT-PCRs (RT-qPCR) have been developed and validated with reference virus isolates and clinical samples. A diagnostic algorithm is proposed for the combined detection in clinical samples and subtyping of SIV strains currently circulating in Europe that is based on a generic, M-gene-specific influenza A virus RT-qPCR. In a second step, positive samples are examined by tetraplex HA- and triplex NA-specific RT-qPCRs to differentiate the porcine subtypes H1, H3, N1 and N2. Within the HA subtype H1, lineages "av" (European avian-derived), "hu" (European human-derived) and "pdm" (human pandemic A/H1N1, 2009) are distinguished by RT-qPCRs, and within the NA subtype N1, lineage "pdm" is differentiated. An RT-PCR amplicon Sanger sequencing method of small fragments of the HA and NA genes is also proposed to safeguard against failure of multiplex RT-qPCR subtyping. These new multiplex RT-qPCR assays provide adequate tools for sustained SIV monitoring programmes in Europe. © 2016 The Authors. Influenza and Other Respiratory Viruses Published by John Wiley & Sons Ltd.

  8. Real-time logo detection and tracking in video

    Science.gov (United States)

    George, M.; Kehtarnavaz, N.; Rahman, M.; Carlsohn, M.

    2010-05-01

    This paper presents a real-time implementation of a logo detection and tracking algorithm in video. The motivation of this work stems from applications on smart phones that require the detection of logos in real-time. For example, one application involves detecting company logos so that customers can easily get special offers in real-time. This algorithm uses a hybrid approach by initially running the Scale Invariant Feature Transform (SIFT) algorithm on the first frame in order to obtain the logo location and then by using an online calibration of color within the SIFT detected area in order to detect and track the logo in subsequent frames in a time efficient manner. The results obtained indicate that this hybrid approach allows robust logo detection and tracking to be achieved in real-time.

  9. Real-Time Detection of a Virus Using Detection Dogs

    Directory of Open Access Journals (Sweden)

    Craig eAngle

    2016-01-01

    Full Text Available Viral infections are ubiquitous in humans, animals, and plants. Real-time methods to identify viral infections are limited and do not exist for use in harsh or resource-constrained environments. Previous research identified that tissues produce unique volatile organic compounds (VOC and demonstrated that VOC concentrations change during pathologic states including infection, neoplasia, or metabolic disease. Patterns of VOC expression may be pathogen-specific and may be associated with an odor that could be used for disease detection.We investigated the ability of two trained dogs to detect cell cultures infected with bovine viral diarrhea virus (BVDV and to discriminate BVDV-infected cell cultures from uninfected cell cultures and from cell cultures infected with bovine herpes virus 1 (BHV 1 and bovine parainfluenza virus 3 (BPIV 3. Dogs were trained to recognize cell cultures infected with two different biotypes of BVDV propagated in MDBK cells using one of three culture media. For detection trials, one target and seven distractors were presented on a scent wheel by a dog handler unaware of the location of targets and distractors.Detection of BVDV- infected cell cultures by Dog 1 had a diagnostic sensitivity of 0.850 (95% CI: 0.701 - 0.942, which was lower than Dog 2 (0.967, 95% CI: 0.837 - 0.994. Both dogs exhibited very high diagnostic specificity (0.981, 95% CI: 0.960 - 0.993 and (0.993, 95% CI: 0.975 - 0.999, respectively.These findings demonstrate that trained dogs can differentiate between cultured cells infected with BVDV, BHV1, and BPIV3 and are a realistic real-time mobile pathogen sensing technology for viral pathogens. The ability to discriminate between target and distractor samples plausibly results from expression of unique VOC patterns virus-infected and uninfected cells.

  10. Real-Time Detection of a Virus Using Detection Dogs.

    Science.gov (United States)

    Angle, T Craig; Passler, Thomas; Waggoner, Paul L; Fischer, Terrence D; Rogers, Bart; Galik, Patricia K; Maxwell, Herris S

    2015-01-01

    Viral infections are ubiquitous in humans, animals, and plants. Real-time methods to identify viral infections are limited and do not exist for use in harsh or resource-constrained environments. Previous research identified that tissues produce unique volatile organic compounds (VOC) and demonstrated that VOC concentrations change during pathologic states, including infection, neoplasia, or metabolic disease. Patterns of VOC expression may be pathogen specific and may be associated with an odor that could be used for disease detection. We investigated the ability of two trained dogs to detect cell cultures infected with bovine viral diarrhea virus (BVDV) and to discriminate BVDV-infected cell cultures from uninfected cell cultures and from cell cultures infected with bovine herpes virus 1 (BHV 1) and bovine parainfluenza virus 3 (BPIV 3). Dogs were trained to recognize cell cultures infected with two different biotypes of BVDV propagated in Madin-Darby bovine kidney cells using one of three culture media. For detection trials, one target and seven distractors were presented on a scent wheel by a dog handler unaware of the location of targets and distractors. Detection of BVDV-infected cell cultures by Dog 1 had a diagnostic sensitivity of 0.850 (95% CI: 0.701-0.942), which was lower than Dog 2 (0.967, 95% CI: 0.837-0.994). Both dogs exhibited very high diagnostic specificity (0.981, 95% CI: 0.960-0.993) and (0.993, 95% CI: 0.975-0.999), respectively. These findings demonstrate that trained dogs can differentiate between cultured cells infected with BVDV, BHV1, and BPIV3 and are a realistic real-time mobile pathogen sensing technology for viral pathogens. The ability to discriminate between target and distractor samples plausibly results from expression of unique VOC patterns in virus-infected and -uninfected cells.

  11. Rapid detection and subtyping of European swine influenza viruses in porcine clinical samples by haemagglutinin- and neuraminidase-specific tetra- and triplex real-time RT-PCRs

    DEFF Research Database (Denmark)

    Henritzi, Dinah; Zhao, Na; Starick, Elke

    2016-01-01

    diagnostic methods which allow for cost-effective large-scale analysis. Methods New SIV haemagglutinin (HA) and neuraminidase (NA) subtype- and lineage-specific multiplex real-time RT-PCRs (RT-qPCR) have been developed and validated with reference virus isolates and clinical samples. Results A diagnostic....... Swine influenza viruses (SIV) are widespread in European domestic pig populations and evolve dynamically. Knowledge regarding occurrence, spread and evolution of potentially zoonotic SIV in Europe is poorly understood. Objectives Efficient SIV surveillance programmes depend on sensitive and specific......Background A diversifying pool of mammalian-adapted influenza A viruses (IAV) with largely unknown zoonotic potential is maintained in domestic swine populations worldwide. The most recent human influenza pandemic in 2009 was caused by a virus with genes originating from IAV isolated from swine...

  12. Real-Time Pore Pressure Detection: Indicators and Improved Methods

    Directory of Open Access Journals (Sweden)

    Jincai Zhang

    2017-01-01

    Full Text Available High uncertainties may exist in the predrill pore pressure prediction in new prospects and deepwater subsalt wells; therefore, real-time pore pressure detection is highly needed to reduce drilling risks. The methods for pore pressure detection (the resistivity, sonic, and corrected d-exponent methods are improved using the depth-dependent normal compaction equations to adapt to the requirements of the real-time monitoring. A new method is proposed to calculate pore pressure from the connection gas or elevated background gas, which can be used for real-time pore pressure detection. The pore pressure detection using the logging-while-drilling, measurement-while-drilling, and mud logging data is also implemented and evaluated. Abnormal pore pressure indicators from the well logs, mud logs, and wellbore instability events are identified and analyzed to interpret abnormal pore pressures for guiding real-time drilling decisions. The principles for identifying abnormal pressure indicators are proposed to improve real-time pore pressure monitoring.

  13. Real-time change detection for countering improvised explosive devices

    NARCIS (Netherlands)

    Wouw, van de D.W.J.M.; Rens, van K.; Lint, van R.H.; Jaspers, Egbert; With, de P.H.N.; Loce, R.P.; Saber, E.

    2014-01-01

    We explore an automatic real-time change detection system to assist military personnel during transport and surveillance, by detection changes in the environment with respect to a previous operation. Such changes may indicate the presence of Improvised Explosive Devices (IEDs), which can then be

  14. Rapid diagnosis of sepsis with TaqMan-Based multiplex real-time PCR.

    Science.gov (United States)

    Liu, Chang-Feng; Shi, Xin-Ping; Chen, Yun; Jin, Ye; Zhang, Bing

    2018-02-01

    The survival rate of septic patients mainly depends on a rapid and reliable diagnosis. A rapid, broad range, specific and sensitive quantitative diagnostic test is the urgent need. Thus, we developed a TaqMan-Based Multiplex real-time PCR assays to identify bloodstream pathogens within a few hours. Primers and TaqMan probes were designed to be complementary to conserved regions in the 16S rDNA gene of different kinds of bacteria. To evaluate accurately, sensitively, and specifically, the known bacteria samples (Standard strains, whole blood samples) are determined by TaqMan-Based Multiplex real-time PCR. In addition, 30 blood samples taken from patients with clinical symptoms of sepsis were tested by TaqMan-Based Multiplex real-time PCR and blood culture. The mean frequency of positive for Multiplex real-time PCR was 96% at a concentration of 100 CFU/mL, and it was 100% at a concentration greater than 1000 CFU/mL. All the known blood samples and Standard strains were detected positively by TaqMan-Based Multiplex PCR, no PCR products were detected when DNAs from other bacterium were used in the multiplex assay. Among the 30 patients with clinical symptoms of sepsis, 18 patients were confirmed positive by Multiplex real-time PCR and seven patients were confirmed positive by blood culture. TaqMan-Based Multiplex real-time PCR assay with highly sensitivity, specificity and broad detection range, is a rapid and accurate method in the detection of bacterial pathogens of sepsis and should have a promising usage in the diagnosis of sepsis. © 2017 Wiley Periodicals, Inc.

  15. Real-time earthquake monitoring: Early warning and rapid response

    Science.gov (United States)

    1991-01-01

    A panel was established to investigate the subject of real-time earthquake monitoring (RTEM) and suggest recommendations on the feasibility of using a real-time earthquake warning system to mitigate earthquake damage in regions of the United States. The findings of the investigation and the related recommendations are described in this report. A brief review of existing real-time seismic systems is presented with particular emphasis given to the current California seismic networks. Specific applications of a real-time monitoring system are discussed along with issues related to system deployment and technical feasibility. In addition, several non-technical considerations are addressed including cost-benefit analysis, public perceptions, safety, and liability.

  16. Rapid and Accurate Detection of Bacteriophage Activity against Escherichia coli O157:H7 by Propidium Monoazide Real-Time PCR

    Directory of Open Access Journals (Sweden)

    Hui Liu

    2014-01-01

    Full Text Available Conventional methods to determine the efficacy of bacteriophage (phage for biocontrol of E. coli require several days, due to the need to culture bacteria. Furthermore, cell surface-attached phage particles may lyse bacterial cells during experiments, leading to an overestimation of phage activity. DNA-based real-time quantitative polymerase chain reaction (qPCR is a fast, sensitive, and highly specific means of enumerating pathogens. However, qPCR may underestimate phage activity due to its inability to distinguish viable from nonviable cells. In this study, we evaluated the suitability of propidium monoazide (PMA, a microbial membrane-impermeable dye that inhibits amplification of extracellular DNA and DNA within dead or membrane-compromised cells as a means of using qPCR to identify only intact E. coli cells that survive phage exposure. Escherichia coli O157:H7 strain R508N and 4 phages (T5-like, T1-like, T4-like, and O1-like were studied. Results compared PMA-qPCR and direct plating and confirmed that PMA could successfully inhibit amplification of DNA from compromised/damaged cells E. coli O157:H7. Compared to PMA-qPCR, direct plating overestimated (P < 0.01 phage efficacy as cell surface-attached phage particles lysed E. coli O157:H7 during the plating process. Treatment of samples with PMA in combination with qPCR can therefore be considered beneficial when assessing the efficacy of bacteriophage for biocontrol of E. coli O157:H7.

  17. Real-time thermal neutron radiographic detection systems

    International Nuclear Information System (INIS)

    Berger, H.; Bracher, D.A.

    1976-01-01

    Systems for real-time detection of thermal neutron images are reviewed. Characteristics of one system are presented; the data include contrast, resolution and speed of response over the thermal neutron intensity range 2.5 10 3 n/cm 2 -sec to 10 7 n/cm 2 -sec

  18. Real-time change detection in data streams with FPGAs

    International Nuclear Information System (INIS)

    Vega, J.; Dormido-Canto, S.; Cruz, T.; Ruiz, M.; Barrera, E.; Castro, R.; Murari, A.; Ochando, M.

    2014-01-01

    Highlights: • Automatic recognition of changes in data streams of multidimensional signals. • Detection algorithm based on testing exchangeability on-line. • Real-time and off-line applicability. • Real-time implementation in FPGAs. - Abstract: The automatic recognition of changes in data streams is useful in both real-time and off-line data analyses. This article shows several effective change-detecting algorithms (based on martingales) and describes their real-time applicability in the data acquisition systems through the use of Field Programmable Gate Arrays (FPGA). The automatic event recognition system is absolutely general and it does not depend on either the particular event to detect or the specific data representation (waveforms, images or multidimensional signals). The developed approach provides good results for change detection in both the temporal evolution of profiles and the two-dimensional spatial distribution of volume emission intensity. The average computation time in the FPGA is 210 μs per profile

  19. Anomaly detection in real-time gross payment data

    NARCIS (Netherlands)

    Triepels, Ron; Daniels, Hennie; Heijmans, R.; Camp, Olivier; Filipe, Joaquim

    2017-01-01

    We discuss how an autoencoder can detect system-level anomalies in a real-time gross settlement system by reconstructing a set of liquidity vectors. A liquidity vector is an aggregated representation of the underlying payment network of a settlement system for a particular time interval.

  20. Real time avalanche detection for high risk areas.

    Science.gov (United States)

    2014-12-01

    Avalanches routinely occur on State Highway 21 (SH21) between Lowman and Stanley, Idaho each winter. The avalanches pose : a threat to the safety of maintenance workers and the traveling public. A real-time avalanche detection system will allow the :...

  1. Development of a real-time fluorescence loop-mediated isothermal amplification assay for rapid and quantitative detection of Fusarium oxysporum f. sp. cubense tropical race 4 in soil.

    Science.gov (United States)

    Zhang, Xin; Zhang, He; Pu, Jinji; Qi, Yanxiang; Yu, Qunfang; Xie, Yixian; Peng, Jun

    2013-01-01

    Fusarium oxysporum f. sp. cubense (Foc), the causal agent of Fusarium wilt (Panama disease), is one of the most devastating diseases of banana (Musa spp.). The Foc tropical race 4 (TR4) is currently known as a major concern in global banana production. No effective resistance is known in Musa to Foc, and no effective measures for controlling Foc once banana plants have been infected in place. Early and accurate detection of Foc TR4 is essential to protect banana industry and guide banana planting. A real-time fluorescence loop-mediated isothermal amplification assay (RealAmp) was developed for the rapid and quantitative detection of Foc TR4 in soil. The detection limit of the RealAmp assay was approximately 0.4 pg/µl plasmid DNA when mixed with extracted soil DNA or 10(3) spores/g of artificial infested soil, and no cross-reaction with other relative pathogens were observed. The RealAmp assay for quantifying genomic DNA of TR4 was confirmed by testing both artificially and naturally infested samples. Quantification of the soil-borne pathogen DNA of Foc TR4 in naturally infested samples was no significant difference compared to classic real-time PCR (P>0.05). Additionally, RealAmp assay was visual with an improved closed-tube visual detection system by adding SYBR Green I fluorescent dye to the inside of the lid prior to amplification, which avoided the inhibitory effects of the stain on DNA amplification and makes the assay more convenient in the field and could thus become a simple, rapid and effective technique that has potential as an alternative tool for the detection and monitoring of Foc TR4 in field, which would be a routine DNA-based testing service for the soil-borne pathogen in South China.

  2. Development of a real-time fluorescence loop-mediated isothermal amplification assay for rapid and quantitative detection of Fusarium oxysporum f. sp. cubense tropical race 4 in soil.

    Directory of Open Access Journals (Sweden)

    Xin Zhang

    Full Text Available Fusarium oxysporum f. sp. cubense (Foc, the causal agent of Fusarium wilt (Panama disease, is one of the most devastating diseases of banana (Musa spp.. The Foc tropical race 4 (TR4 is currently known as a major concern in global banana production. No effective resistance is known in Musa to Foc, and no effective measures for controlling Foc once banana plants have been infected in place. Early and accurate detection of Foc TR4 is essential to protect banana industry and guide banana planting. A real-time fluorescence loop-mediated isothermal amplification assay (RealAmp was developed for the rapid and quantitative detection of Foc TR4 in soil. The detection limit of the RealAmp assay was approximately 0.4 pg/µl plasmid DNA when mixed with extracted soil DNA or 10(3 spores/g of artificial infested soil, and no cross-reaction with other relative pathogens were observed. The RealAmp assay for quantifying genomic DNA of TR4 was confirmed by testing both artificially and naturally infested samples. Quantification of the soil-borne pathogen DNA of Foc TR4 in naturally infested samples was no significant difference compared to classic real-time PCR (P>0.05. Additionally, RealAmp assay was visual with an improved closed-tube visual detection system by adding SYBR Green I fluorescent dye to the inside of the lid prior to amplification, which avoided the inhibitory effects of the stain on DNA amplification and makes the assay more convenient in the field and could thus become a simple, rapid and effective technique that has potential as an alternative tool for the detection and monitoring of Foc TR4 in field, which would be a routine DNA-based testing service for the soil-borne pathogen in South China.

  3. Reduction of the use of antimicrobial drugs following the rapid detection of Streptococcus agalactiae in the vagina at delivery by real-time PCR assay.

    Science.gov (United States)

    Poncelet-Jasserand, E; Forges, F; Varlet, M-N; Chauleur, C; Seffert, P; Siani, C; Pozzetto, B; Ros, A

    2013-08-01

    To assess whether the determination of the presence of group B streptococci (GBS) in the vagina using a rapid polymerase chain reaction (PCR) assay at delivery was able to spare useless antimicrobial treatments, as compared with conventional culture at 34-38 weeks of gestation. Practical evaluation and prospective cost-effectiveness analysis. A university hospital in France. A cohort of 225 women in labour at the University-Hospital of Saint-Etienne. Each woman had a conventional culture performed at 34-38 weeks of gestation. At the beginning of labour, two vaginal swabs were sampled for rapid PCR testing and culture. The decision to prescribe a prophylactic antimicrobial treatment or not was taken according to the result of the PCR test. A comparative cost-effectiveness analysis of the two diagnostic strategies was carried out. Number of women receiving inadequate prophylactic antimicrobial drugs following each testing strategy, costs of PCR testing and culture, frequency of vaginal GBS, and diagnostic performance of the PCR test at delivery. The percentage of unnecessarily treated women was significantly reduced using the rapid test versus conventional culture (4.5 and 13.6%, respectively; P < 0.001). The rate of vaginal GBS at delivery was 12.5%. The incremental cost-effectiveness ratio (ICER) for each inadequate management avoided was €36 and €173 from the point of view of the healthcare system and hospital, respectively. The PCR assay reduced the number of inadequate antimicrobial treatments aimed to prevent the early onset of GBS disease. However, this strategy generates extra costs that must be put into balance with its clinical benefits. © 2013 The Authors BJOG An International Journal of Obstetrics and Gynaecology © 2013 RCOG.

  4. Real-time Multiple Abnormality Detection in Video Data

    DEFF Research Database (Denmark)

    Have, Simon Hartmann; Ren, Huamin; Moeslund, Thomas B.

    2013-01-01

    Automatic abnormality detection in video sequences has recently gained an increasing attention within the research community. Although progress has been seen, there are still some limitations in current research. While most systems are designed at detecting specific abnormality, others which...... are capable of detecting more than two types of abnormalities rely on heavy computation. Therefore, we provide a framework for detecting abnormalities in video surveillance by using multiple features and cascade classifiers, yet achieve above real-time processing speed. Experimental results on two datasets...... show that the proposed framework can reliably detect abnormalities in the video sequence, outperforming the current state-of-the-art methods....

  5. Frame based Motion Detection for real-time Surveillance

    OpenAIRE

    Brajesh Patel; Neelam Patel

    2012-01-01

    In this paper a series of algorithm has been formed to track the feature of motion detection under surveillance system. In the proposed work a pixel variant plays a vital role in detection of moving object of a particular clip. If there is a little bit motion in a frame then it is detected very easily by calculating pixel variance. This algorithm detects the zero variation only when there is no motion in a real-time video sequence. It is simple and easier for motion detection in the fames of ...

  6. Real-Time Analytics for the Healthcare Industry: Arrhythmia Detection.

    Science.gov (United States)

    Agneeswaran, Vijay Srinivas; Mukherjee, Joydeb; Gupta, Ashutosh; Tonpay, Pranay; Tiwari, Jayati; Agarwal, Nitin

    2013-09-01

    It is time for the healthcare industry to move from the era of "analyzing our health history" to the age of "managing the future of our health." In this article, we illustrate the importance of real-time analytics across the healthcare industry by providing a generic mechanism to reengineer traditional analytics expressed in the R programming language into Storm-based real-time analytics code. This is a powerful abstraction, since most data scientists use R to write the analytics and are not clear on how to make the data work in real-time and on high-velocity data. Our paper focuses on the applications necessary to a healthcare analytics scenario, specifically focusing on the importance of electrocardiogram (ECG) monitoring. A physician can use our framework to compare ECG reports by categorization and consequently detect Arrhythmia. The framework can read the ECG signals and uses a machine learning-based categorizer that runs within a Storm environment to compare different ECG signals. The paper also presents some performance studies of the framework to illustrate the throughput and accuracy trade-off in real-time analytics.

  7. [Real-time PCR in rapid diagnosis of Aeromonas hydrophila necrotizing soft tissue infections].

    Science.gov (United States)

    Kohayagawa, Yoshitaka; Izumi, Yoko; Ushita, Misuzu; Niinou, Norio; Koshizaki, Masayuki; Yamamori, Yuji; Kaneko, Sakae; Fukushima, Hiroshi

    2009-11-01

    We report a case of rapidly progressive necrotizing soft tissue infection and sepsis followed by a patient's death. We suspected Vibrio vulnificus infection because the patient's underlying disease was cirrhosis and the course extremely rapid. No microbe had been detected at death. We extracted DNA from a blood culture bottle. SYBR green I real-time PCR was conducted but could not detect V. vulnificus vvh in the DNA sample. Aeromonas hydrophila was cultured and identified in blood and necrotized tissue samples. Real-time PCR was conducted to detect A. hydrophila ahh1, AHCYTOEN and aerA in the DNA sample extracted from the blood culture bottle and an isolated necrotized tissue strain, but only ahh1 was positive. High-mortality in necrotizing soft tissue infections makes it is crucial to quickly detect V. vulnificus and A. hydrophila. We found real-time PCR for vvh, ahh1, AHCYTOEN, and aerA useful in detecting V. vulnificus and A. hydrophila in necrotizing soft tissue infections.

  8. Wide area surveillance real-time motion detection systems

    CERN Document Server

    2014-01-01

    The book describes a system for visual surveillance using intelligent cameras. The camera uses robust techniques for detecting and tracking moving objects. The real time capture of the objects is then stored int he database. The tracking data stored in the database is analysed to study the camera view, detect and track objects, and study object behavior. These set of models provide a robust framework for coordinating the tracking of objects between overlapping and non-overlapping cameras, and recording the activity of objects detected by the system.

  9. A Real-Time PCR Assay Based on 5.8S rRNA Gene (5.8S rDNA) for Rapid Detection of Candida from Whole Blood Samples.

    Science.gov (United States)

    Guo, Yi; Yang, Jing-Xian; Liang, Guo-Wei

    2016-06-01

    The prevalence of Candida in bloodstream infections (BSIs) has increased. To date, the identification of Candida in BSIs still mainly relies on blood culture and serological tests, but they have various limitations. Therefore, a real-time PCR assay for the detection of Candida from whole blood is presented. The unique primers/probe system was designed on 5.8S rRNA gene (5.8S rDNA) of Candida genus. The analytical sensitivity was determined by numbers of positive PCRs in 12 repetitions. At the concentration of 10(1) CFU/ml blood, positive PCR rates of 100 % were obtained for C. albicans, C. parapsilosis, C. tropicalis, and C. krusei. The detection rate for C. glabrata was 75 % at 10(1) CFU/ml blood. The reaction specificity was 100 % when evaluating the assay using DNA samples from clinical isolates and human blood. The maximum CVs of intra-assay and inter-assay for the detection limit were 1.22 and 2.22 %, respectively. To assess the clinical applicability, 328 blood samples from 82 patients were prospectively tested and real-time PCR results were compared with results from blood culture. Diagnostic sensitivity of the PCR was 100 % using as gold standard blood culture, and specificity was 98.4 %. Our data suggest that the developed assay can be used in clinical laboratories as an accurate and rapid screening test for the Candida from whole blood. Although further evaluation is warranted, our assay holds promise for earlier diagnosis of candidemia.

  10. Shape based kinetic outlier detection in real-time PCR

    Directory of Open Access Journals (Sweden)

    D'Atri Mario

    2010-04-01

    Full Text Available Abstract Background Real-time PCR has recently become the technique of choice for absolute and relative nucleic acid quantification. The gold standard quantification method in real-time PCR assumes that the compared samples have similar PCR efficiency. However, many factors present in biological samples affect PCR kinetic, confounding quantification analysis. In this work we propose a new strategy to detect outlier samples, called SOD. Results Richards function was fitted on fluorescence readings to parameterize the amplification curves. There was not a significant correlation between calculated amplification parameters (plateau, slope and y-coordinate of the inflection point and the Log of input DNA demonstrating that this approach can be used to achieve a "fingerprint" for each amplification curve. To identify the outlier runs, the calculated parameters of each unknown sample were compared to those of the standard samples. When a significant underestimation of starting DNA molecules was found, due to the presence of biological inhibitors such as tannic acid, IgG or quercitin, SOD efficiently marked these amplification profiles as outliers. SOD was subsequently compared with KOD, the current approach based on PCR efficiency estimation. The data obtained showed that SOD was more sensitive than KOD, whereas SOD and KOD were equally specific. Conclusion Our results demonstrated, for the first time, that outlier detection can be based on amplification shape instead of PCR efficiency. SOD represents an improvement in real-time PCR analysis because it decreases the variance of data thus increasing the reliability of quantification.

  11. Real-time door detection for indoor autonomous vehicle

    Science.gov (United States)

    He, Zhihao; Zhu, Ming

    2017-07-01

    Indoor Autonomous Vehicle(IAV) is used in many indoor scenes. Such as hotels and hospitals. Door detection is a key issue to guide the IAV into rooms. In this paper, we consider door detection in the use of indoor navigation of IAV. Since real-time properties are important for real-world IAV, the detection algorithm must be fast enough. Most monocular-camera based door detection model need a perfect detection of the four line segments of the door or the four corners. But in many situations, line segments could be extended or cut off. And there could be many false detected corners. And few of them can distinguish doors from door-like objects with door-like shape effectively. We proposed a 2-D vision model of the door that is made up of line segments. The number of parts detected is used to determine the possibility of a door. Our algorithm is tested on a database of doors.1 The robustness and real-time are verified. The precision is 89.4%. Average time consumed for processing a 640x320 figure is 44.73ms.

  12. Real-time Face Detection using Skin Color Model

    Institute of Scientific and Technical Information of China (English)

    LU Yao-xin; LIU Zhi-Qiang; ZHU Xiang-hua

    2004-01-01

    This paper presents a new face detection approach to real-time applications, which is based on the skin color model and the morphological filtering. First the non-skin color pixels of the input image are removed based on the skin color model in the YCrCb chrominance space, from which we extract candidate human face regions. Then a mathematical morphological filter is used to remove noisy regions and fill the holes in the candidate skin color regions. We adopt the similarity between the human face features and the candidate face regions to locate the face regions in the original image. We have implemented the algorithm in our smart media system. The experiment results show that this system is effective in real-time applications.

  13. Aircraft Fault Detection Using Real-Time Frequency Response Estimation

    Science.gov (United States)

    Grauer, Jared A.

    2016-01-01

    A real-time method for estimating time-varying aircraft frequency responses from input and output measurements was demonstrated. The Bat-4 subscale airplane was used with NASA Langley Research Center's AirSTAR unmanned aerial flight test facility to conduct flight tests and collect data for dynamic modeling. Orthogonal phase-optimized multisine inputs, summed with pilot stick and pedal inputs, were used to excite the responses. The aircraft was tested in its normal configuration and with emulated failures, which included a stuck left ruddervator and an increased command path latency. No prior knowledge of a dynamic model was used or available for the estimation. The longitudinal short period dynamics were investigated in this work. Time-varying frequency responses and stability margins were tracked well using a 20 second sliding window of data, as compared to a post-flight analysis using output error parameter estimation and a low-order equivalent system model. This method could be used in a real-time fault detection system, or for other applications of dynamic modeling such as real-time verification of stability margins during envelope expansion tests.

  14. Microcontroller-based real-time QRS detection.

    Science.gov (United States)

    Sun, Y; Suppappola, S; Wrublewski, T A

    1992-01-01

    The authors describe the design of a system for real-time detection of QRS complexes in the electrocardiogram based on a single-chip microcontroller (Motorola 68HC811). A systematic analysis of the instrumentation requirements for QRS detection and of the various design techniques is also given. Detection algorithms using different nonlinear transforms for the enhancement of QRS complexes are evaluated by using the ECG database of the American Heart Association. The results show that the nonlinear transform involving multiplication of three adjacent, sign-consistent differences in the time domain gives a good performance and a quick response. When implemented with an appropriate sampling rate, this algorithm is also capable of rejecting pacemaker spikes. The eight-bit single-chip microcontroller provides sufficient throughput and shows a satisfactory performance. Implementation of multiple detection algorithms in the same system improves flexibility and reliability. The low chip count in the design also favors maintainability and cost-effectiveness.

  15. Real-time driver fatigue detection based on face alignment

    Science.gov (United States)

    Tao, Huanhuan; Zhang, Guiying; Zhao, Yong; Zhou, Yi

    2017-07-01

    The performance and robustness of fatigue detection largely decrease if the driver with glasses. To address this issue, this paper proposes a practical driver fatigue detection method based on face alignment at 3000 FPS algorithm. Firstly, the eye regions of the driver are localized by exploiting 6 landmarks surrounding each eye. Secondly, the HOG features of the extracted eye regions are calculated and put into SVM classifier to recognize the eye state. Finally, the value of PERCLOS is calculated to determine whether the driver is drowsy or not. An alarm will be generated if the eye is closed for a specified period of time. The accuracy and real-time on testing videos with different drivers demonstrate that the proposed algorithm is robust and obtain better accuracy for driver fatigue detection compared with some previous method.

  16. Aggregated channels network for real-time pedestrian detection

    Science.gov (United States)

    Ghorban, Farzin; Marín, Javier; Su, Yu; Colombo, Alessandro; Kummert, Anton

    2018-04-01

    Convolutional neural networks (CNNs) have demonstrated their superiority in numerous computer vision tasks, yet their computational cost results prohibitive for many real-time applications such as pedestrian detection which is usually performed on low-consumption hardware. In order to alleviate this drawback, most strategies focus on using a two-stage cascade approach. Essentially, in the first stage a fast method generates a significant but reduced amount of high quality proposals that later, in the second stage, are evaluated by the CNN. In this work, we propose a novel detection pipeline that further benefits from the two-stage cascade strategy. More concretely, the enriched and subsequently compressed features used in the first stage are reused as the CNN input. As a consequence, a simpler network architecture, adapted for such small input sizes, allows to achieve real-time performance and obtain results close to the state-of-the-art while running significantly faster without the use of GPU. In particular, considering that the proposed pipeline runs in frame rate, the achieved performance is highly competitive. We furthermore demonstrate that the proposed pipeline on itself can serve as an effective proposal generator.

  17. Real-Time EEG-Based Happiness Detection System

    Directory of Open Access Journals (Sweden)

    Noppadon Jatupaiboon

    2013-01-01

    Full Text Available We propose to use real-time EEG signal to classify happy and unhappy emotions elicited by pictures and classical music. We use PSD as a feature and SVM as a classifier. The average accuracies of subject-dependent model and subject-independent model are approximately 75.62% and 65.12%, respectively. Considering each pair of channels, temporal pair of channels (T7 and T8 gives a better result than the other area. Considering different frequency bands, high-frequency bands (Beta and Gamma give a better result than low-frequency bands. Considering different time durations for emotion elicitation, that result from 30 seconds does not have significant difference compared with the result from 60 seconds. From all of these results, we implement real-time EEG-based happiness detection system using only one pair of channels. Furthermore, we develop games based on the happiness detection system to help user recognize and control the happiness.

  18. Development and evaluation of a real-time fluorogenic loop-mediated isothermal amplification assay integrated on a microfluidic disc chip (on-chip LAMP) for rapid and simultaneous detection of ten pathogenic bacteria in aquatic animals.

    Science.gov (United States)

    Zhou, Qian-Jin; Wang, Lei; Chen, Jiong; Wang, Rui-Na; Shi, Yu-Hong; Li, Chang-Hong; Zhang, De-Min; Yan, Xiao-Jun; Zhang, Yan-Jun

    2014-09-01

    Rapid, low-cost, and user-friendly strategies are urgently needed for early disease diagnosis and timely treatment, particularly for on-site screening of pathogens in aquaculture. In this study, we successfully developed a real-time fluorogenic loop-mediated isothermal amplification assay integrated on a microfluidic disc chip (on-chip LAMP), which was capable of simultaneously detecting 10 pathogenic bacteria in aquatic animals, i.e., Nocardia seriolae, Pseudomonas putida, Streptococcus iniae, Vibrio alginolyticus, Vibrio anguillarum, Vibrio fluvialis, Vibrio harveyi, Vibrio parahaemolyticus, Vibrio rotiferianus, and Vibrio vulnificus. The assay provided a nearly-automated approach, with only a single pipetting step per chip for sample dispensing. This technique could achieve limits of detection (LOD) ranging from 0.40 to 6.42pg per 1.414μL reaction in less than 30 min. The robust reproducibility was demonstrated by a little variation among duplications for each bacterium with the coefficient of variation (CV) for time to positive (Tp) value less than 0.10. The clinical sensitivity and specificity of this on-chip LAMP assay in detecting field samples were 96.2% and 93.8% by comparison with conventional microbiological methods. Compared with other well-known techniques, on-chip LAMP assay provides low sample and reagent consumption, ease-of-use, accelerated analysis, multiple bacteria and on-site detection, and high reproducibility, indicating that such a technique would be applicable for on-site detection and routine monitoring of multiple pathogens in aquaculture. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Development of a real-time quantitative assay for detection of Epstein-Barr virus

    NARCIS (Netherlands)

    Niesters, H. G.; van Esser, J.; Fries, E.; Wolthers, K. C.; Cornelissen, J.; Osterhaus, A. D.

    2000-01-01

    With the use of real-time PCR, we developed and evaluated a rapid, sensitive, specific, and reproducible method for the detection of Epstein-Barr virus (EBV) DNA in plasma samples. This method allowed us to screen plasma and serum samples over a range between 100 and 10(7) copies of DNA per ml using

  20. Development of a real-time quantitative assay for detection of Epstein-Barr virus

    NARCIS (Netherlands)

    H.G.M. Niesters (Bert); E. Fries; K.C. Wolthers (Katja); A.D.M.E. Osterhaus (Albert); J.J. Cornelissen (Jan); J.W.J. van Esser (Joost)

    2000-01-01

    textabstractWith the use of real-time PCR, we developed and evaluated a rapid, sensitive, specific, and reproducible method for the detection of Epstein-Barr virus (EBV) DNA in plasma samples. This method allowed us to screen plasma and serum samples over a range between 100 and

  1. Real-time people and vehicle detection from UAV imagery

    Science.gov (United States)

    Gaszczak, Anna; Breckon, Toby P.; Han, Jiwan

    2011-01-01

    A generic and robust approach for the real-time detection of people and vehicles from an Unmanned Aerial Vehicle (UAV) is an important goal within the framework of fully autonomous UAV deployment for aerial reconnaissance and surveillance. Here we present an approach for the automatic detection of vehicles based on using multiple trained cascaded Haar classifiers with secondary confirmation in thermal imagery. Additionally we present a related approach for people detection in thermal imagery based on a similar cascaded classification technique combining additional multivariate Gaussian shape matching. The results presented show the successful detection of vehicle and people under varying conditions in both isolated rural and cluttered urban environments with minimal false positive detection. Performance of the detector is optimized to reduce the overall false positive rate by aiming at the detection of each object of interest (vehicle/person) at least once in the environment (i.e. per search patter flight path) rather than every object in each image frame. Currently the detection rate for people is ~70% and cars ~80% although the overall episodic object detection rate for each flight pattern exceeds 90%.

  2. Optical sensor for real-time weld defect detection

    Science.gov (United States)

    Ancona, Antonio; Maggipinto, Tommaso; Spagnolo, Vincenzo; Ferrara, Michele; Lugara, Pietro M.

    2002-04-01

    In this work we present an innovative optical sensor for on- line and non-intrusive welding process monitoring. It is based on the spectroscopic analysis of the optical VIS emission of the welding plasma plume generated in the laser- metal interaction zone. Plasma electron temperature has been measured for different chemical species composing the plume. Temperature signal evolution has been recorded and analyzed during several CO2-laser welding processes, under variable operating conditions. We have developed a suitable software able to real time detect a wide range of weld defects like crater formation, lack of fusion, excessive penetration, seam oxidation. The same spectroscopic approach has been applied for electric arc welding process monitoring. We assembled our optical sensor in a torch for manual Gas Tungsten Arc Welding procedures and tested the prototype in a manufacturing industry production line. Even in this case we found a clear correlation between the signal behavior and the welded joint quality.

  3. A rapid and direct real time PCR-based method for identification of Salmonella spp

    DEFF Research Database (Denmark)

    Rodriguez-Lazaro, D.; Hernández, Marta; Esteve, T.

    2003-01-01

    The aim of this work was the validation of a rapid, real-time PCR assay based on TaqMan((R)) technology for the unequivocal identification of Salmonella spp. to be used directly on an agar-grown colony. A real-time PCR system targeting at the Salmonella spp. invA gene was optimized and validated ...

  4. A Real-Time PCR Detection of Genus Salmonella in Meat and Milk Samples

    Directory of Open Access Journals (Sweden)

    Jaroslav Pochop

    2013-05-01

    Full Text Available The aim of this study was follow the contamination of ready to eat milk and meat products with Salmonella spp. by using the Step One real-time PCR. Classical microbiological methods for detection of food-borne bacteria involve the use of pre-enrichment and/or specific enrichment, followed by the isolation of the bacteria in solid media and a final confirmation by biochemical and/or serological tests. We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and SensiFAST SYBR Hi-ROX Kit for the real-time PCR performance. In the investigated samples without incubation we could detect strain of Salmonella sp. in five out of twenty three samples (swabs. This Step One real-time PCR assay is extremely useful for any laboratory in possession of a real-time PCR. It is a fast, reproducible, simple, specific and sensitive way to detect nucleic acids, which could be used in clinical diagnostic tests in the future. Our results indicated that the Step One real-time PCR assay developed in this study could sensitively detect Salmonella spp. in ready to eat food.

  5. Real-time centre detection of an OLED structure

    NARCIS (Netherlands)

    Pieters, R.S.; Jonker, P.P.; Nijmeijer, H.

    2009-01-01

    The research presented in this paper focuses on real-time image processing for visual servoing, i.e. the positioning of a x-y table by using a camera only instead of encoders. A camera image stream plus real-time image processing determines the position in the next iteration of the table controller.

  6. Real-Time Center Detection of an OLED Structure

    NARCIS (Netherlands)

    Pieters, R.S.; Jonker, P.P.; Nijmeijer, H.; Blanc-Talon, J.; Philips, W.; Popescu, D.; Scheunders, P.

    2009-01-01

    The research presented in this paper focuses on real-time image processing for visual servoing, i.e. the positioning of a x-y table by using a camera only instead of encoders. A camera image stream plus real-time image processing determines the position in the next iteration of the table controller.

  7. Study on APD real time compensation methods of laser Detection system

    International Nuclear Information System (INIS)

    Feng Ying; Zhang He; Zhang Xiangjin; Liu Kun

    2011-01-01

    their operating principles. The constant false alarm rate compensation can't detect the pulse signal which comes randomly. Therefore real-time performance can't be realized. The noise compensation can meet the request of real-time performance. If it is used in the environment where background light is intense or changes acutely, there is a better effect. The temperature compensation can also achieve the real-time performance request. If it is used in the environment where temperature changes acutely, there is also a better effect. Aim at such problems, this paper presents that different APD real-time compensations should be adopt to adapt to different environments. The exiting temperature compensation adjusts output voltage by using variable resistance to regulate input voltage. Its structure is complex; the real-time performance is worse. In order to remedy these defects, a real-time temperature compensation which is based on switch on-off time of switching power supply is designed. Its feasibility and operating stability are confirmed by plate making and experiment. At last, the comparison experiments between the real-time noise compensation and the real-time temperature compensation is carried out in the environments where temperature is almost invariant and background light acutely changes from5lux to150lux . The result shows that the operating effect of the real-time noise compensation is better here, the noise minifies to a sixth of original noise. The comparison experiments between the real-time noise compensation and the real-time temperature compensation is carried out in darkroom where background light is 5lux and temperature almost rapidly changes from -20 deg. C to 80 deg. C. The result shows that the operating effect of the real-time temperature compensation is better here, the noise minifies to a seventh of original noise. Moreover, these methods can be applied to other type detection systems of weak photoelectric signal; they have high actual application

  8. Study on APD real time compensation methods of laser Detection system

    Energy Technology Data Exchange (ETDEWEB)

    Feng Ying; Zhang He; Zhang Xiangjin; Liu Kun, E-mail: fy_caimi@163.com [ZNDY of Ministerial Key Laboratory, Nanjing University of Science and Technology, Nanjing 210094 (China)

    2011-02-01

    by analyzing their operating principles. The constant false alarm rate compensation can't detect the pulse signal which comes randomly. Therefore real-time performance can't be realized. The noise compensation can meet the request of real-time performance. If it is used in the environment where background light is intense or changes acutely, there is a better effect. The temperature compensation can also achieve the real-time performance request. If it is used in the environment where temperature changes acutely, there is also a better effect. Aim at such problems, this paper presents that different APD real-time compensations should be adopt to adapt to different environments. The exiting temperature compensation adjusts output voltage by using variable resistance to regulate input voltage. Its structure is complex; the real-time performance is worse. In order to remedy these defects, a real-time temperature compensation which is based on switch on-off time of switching power supply is designed. Its feasibility and operating stability are confirmed by plate making and experiment. At last, the comparison experiments between the real-time noise compensation and the real-time temperature compensation is carried out in the environments where temperature is almost invariant and background light acutely changes from5lux to150lux . The result shows that the operating effect of the real-time noise compensation is better here, the noise minifies to a sixth of original noise. The comparison experiments between the real-time noise compensation and the real-time temperature compensation is carried out in darkroom where background light is 5lux and temperature almost rapidly changes from -20 deg. C to 80 deg. C. The result shows that the operating effect of the real-time temperature compensation is better here, the noise minifies to a seventh of original noise. Moreover, these methods can be applied to other type detection systems of weak photoelectric signal; they

  9. Study on APD real time compensation methods of laser Detection system

    Science.gov (United States)

    Ying, Feng; He, Zhang; Xiangjin, Zhang; Kun, Liu

    2011-02-01

    their operating principles. The constant false alarm rate compensation can't detect the pulse signal which comes randomly. Therefore real-time performance can't be realized. The noise compensation can meet the request of real-time performance. If it is used in the environment where background light is intense or changes acutely, there is a better effect. The temperature compensation can also achieve the real-time performance request. If it is used in the environment where temperature changes acutely, there is also a better effect. Aim at such problems, this paper presents that different APD real-time compensations should be adopt to adapt to different environments. The exiting temperature compensation adjusts output voltage by using variable resistance to regulate input voltage. Its structure is complex; the real-time performance is worse. In order to remedy these defects, a real-time temperature compensation which is based on switch on-off time of switching power supply is designed. Its feasibility and operating stability are confirmed by plate making and experiment. At last, the comparison experiments between the real-time noise compensation and the real-time temperature compensation is carried out in the environments where temperature is almost invariant and background light acutely changes from5lux to150lux . The result shows that the operating effect of the real-time noise compensation is better here, the noise minifies to a sixth of original noise. The comparison experiments between the real-time noise compensation and the real-time temperature compensation is carried out in darkroom where background light is 5lux and temperature almost rapidly changes from -20°C to 80°C. The result shows that the operating effect of the real-time temperature compensation is better here, the noise minifies to a seventh of original noise. Moreover, these methods can be applied to other type detection systems of weak photoelectric signal; they have high actual application value.

  10. Real-Time Rotational Activity Detection in Atrial Fibrillation

    Science.gov (United States)

    Ríos-Muñoz, Gonzalo R.; Arenal, Ángel; Artés-Rodríguez, Antonio

    2018-01-01

    Rotational activations, or spiral waves, are one of the proposed mechanisms for atrial fibrillation (AF) maintenance. We present a system for assessing the presence of rotational activity from intracardiac electrograms (EGMs). Our system is able to operate in real-time with multi-electrode catheters of different topologies in contact with the atrial wall, and it is based on new local activation time (LAT) estimation and rotational activity detection methods. The EGM LAT estimation method is based on the identification of the highest sustained negative slope of unipolar signals. The method is implemented as a linear filter whose output is interpolated on a regular grid to match any catheter topology. Its operation is illustrated on selected signals and compared to the classical Hilbert-Transform-based phase analysis. After the estimation of the LAT on the regular grid, the detection of rotational activity in the atrium is done by a novel method based on the optical flow of the wavefront dynamics, and a rotation pattern match. The methods have been validated using in silico and real AF signals. PMID:29593566

  11. REAL-TIME PCR DETECTION OF LISTERIA MONOCYTOGENES IN FOOD SAMPLES OF ANIMAL ORIGIN

    Directory of Open Access Journals (Sweden)

    Jaroslav Pochop

    2013-02-01

    Full Text Available The aim of this study was to follow the contamination of food with Listeria monocytogenes by using Step One real time polymerase chain reaction (PCR. We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and SensiFAST SYBR Hi-ROX Kit for the real-time PCR performance. In 24 samples of food of animal origin without incubation were detected strains of Listeria monocytogenes in 15 samples (swabs. Nine samples were negative. Our results indicated that the real-time PCR assay developed in this study could sensitively detect Listeria monocytogenes in food of animal origin without incubation. This could prevent infection caused by Listeria monocytogenes, and also could benefit food manufacturing companies by extending their product’s shelf-life as well as saving the cost of warehousing their food products while awaiting pathogen testing results. The rapid real-time PCR-based method performed very well compared to the conventional method. It is a fast, simple, specific and sensitive way to detect nucleic acids, which could be used in clinical diagnostic tests in the future.

  12. Robust real-time change detection in high jitter.

    Energy Technology Data Exchange (ETDEWEB)

    Simonson, Katherine Mary; Ma, Tian J.

    2009-08-01

    A new method is introduced for real-time detection of transient change in scenes observed by staring sensors that are subject to platform jitter, pixel defects, variable focus, and other real-world challenges. The approach uses flexible statistical models for the scene background and its variability, which are continually updated to track gradual drift in the sensor's performance and the scene under observation. Two separate models represent temporal and spatial variations in pixel intensity. For the temporal model, each new frame is projected into a low-dimensional subspace designed to capture the behavior of the frame data over a recent observation window. Per-pixel temporal standard deviation estimates are based on projection residuals. The second approach employs a simple representation of jitter to generate pixelwise moment estimates from a single frame. These estimates rely on spatial characteristics of the scene, and are used gauge each pixel's susceptibility to jitter. The temporal model handles pixels that are naturally variable due to sensor noise or moving scene elements, along with jitter displacements comparable to those observed in the recent past. The spatial model captures jitter-induced changes that may not have been seen previously. Change is declared in pixels whose current values are inconsistent with both models.

  13. Facial Expression Emotion Detection for Real-Time Embedded Systems

    Directory of Open Access Journals (Sweden)

    Saeed Turabzadeh

    2018-01-01

    Full Text Available Recently, real-time facial expression recognition has attracted more and more research. In this study, an automatic facial expression real-time system was built and tested. Firstly, the system and model were designed and tested on a MATLAB environment followed by a MATLAB Simulink environment that is capable of recognizing continuous facial expressions in real-time with a rate of 1 frame per second and that is implemented on a desktop PC. They have been evaluated in a public dataset, and the experimental results were promising. The dataset and labels used in this study were made from videos, which were recorded twice from five participants while watching a video. Secondly, in order to implement in real-time at a faster frame rate, the facial expression recognition system was built on the field-programmable gate array (FPGA. The camera sensor used in this work was a Digilent VmodCAM — stereo camera module. The model was built on the Atlys™ Spartan-6 FPGA development board. It can continuously perform emotional state recognition in real-time at a frame rate of 30. A graphical user interface was designed to display the participant’s video in real-time and two-dimensional predict labels of the emotion at the same time.

  14. Development of a highly sensitive one-tube nested real-time PCR for detecting Mycobacterium tuberculosis.

    Science.gov (United States)

    Choi, Yeonim; Jeon, Bo-Young; Shim, Tae Sun; Jin, Hyunwoo; Cho, Sang-Nae; Lee, Hyeyoung

    2014-12-01

    Rapid, accurate detection of Mycobacterium tuberculosis is crucial in the diagnosis of tuberculosis (TB), but conventional diagnostic methods have limited sensitivity and specificity or are time consuming. A new highly sensitive nucleic acid amplification test, combined nested and real-time polymerase chain reaction (PCR) in a single tube (one-tube nested real-time PCR), was developed for detecting M. tuberculosis, which takes advantage of two PCR techniques, i.e., nested PCR and real-time PCR. One-tube nested real-time PCR was designed to have two sequential reactions with two sets of primers and dual probes for the insertion sequence (IS) 6110 sequence of M. tuberculosis in a single closed tube. The minimum limits of detection of IS6110 real-time PCR and IS6110 one-tube nested real-time PCR were 100 fg/μL and 1 fg/μL of M. tuberculosis DNA, respectively. AdvanSure TB/non-tuberculous mycobacteria (NTM) real-time PCR, IS6110 real-time PCR, and two-tube nested real-time PCR showed 100% sensitivity and 100% specificity for clinical M. tuberculosis isolates and NTM isolates. In comparison, the sensitivities of AdvanSure TB/NTM real-time PCR, single IS6110 real-time PCR, and one-tube nested real-time PCR were 91% (152/167), 94.6% (158/167), and 100% (167/167) for sputum specimens, respectively. In conclusion, IS6110 one-tube nested real-time PCR is useful for detecting M. tuberculosis due to its high sensitivity and simple manipulation. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Detection of Balamuthia mandrillaris DNA by real-time PCR targeting the RNase P gene

    Directory of Open Access Journals (Sweden)

    Lewin Astrid

    2008-12-01

    Full Text Available Abstract Background The free-living amoeba Balamuthia mandrillaris may cause fatal encephalitis both in immunocompromised and in – apparently – immunocompetent humans and other mammalian species. Rapid, specific, sensitive, and reliable detection requiring little pathogen-specific expertise is an absolute prerequisite for a successful therapy and a welcome tool for both experimental and epidemiological research. Results A real-time polymerase chain reaction assay using TaqMan® probes (real-time PCR was established specifically targeting the RNase P gene of B. mandrillaris amoebae. The assay detected at least 2 (down to 0.5 genomes of B. mandrillaris grown in axenic culture. It did not react with DNA from closely related Acanthamoeba (3 species, nor with DNA from Toxoplasma gondii, Leishmania major, Pneumocystis murina, Mycobacterium bovis (BCG, human brain, various mouse organs, or from human and murine cell lines. The assay efficiently detected B. mandrillaris DNA in spiked cell cultures, spiked murine organ homogenates, B. mandrillaris-infected mice, and CNS tissue-DNA preparations from 2 patients with proven cerebral balamuthiasis. This novel primer set was successfully combined with a published set that targets the B. mandrillaris 18S rRNA gene in a duplex real-time PCR assay to ensure maximum specificity and as a precaution against false negative results. Conclusion A real-time PCR assay for B. mandrillaris amoebae is presented, that is highly specific, sensitive, and reliable and thus suited both for diagnosis and for research.

  16. UMTS rapid response real-time seismic networks: implementation and strategies at INGV

    Science.gov (United States)

    Govoni, Aladino; Margheriti, Lucia; Moretti, Milena; Lauciani, Valentino; Sensale, Gianpaolo; Bucci, Augusto; Criscuoli, Fabio

    2015-04-01

    The benefits of portable real-time seismic networks are several and well known. During the management of a temporary experiment from the real-time data it is possible to detect and fix rapidly problems with power supply, time synchronization, disk failures and, most important, seismic signal quality degradation due to unexpected noise sources or sensor alignment/tampering. This usually minimizes field maintenance trips and maximizes both the quantity and the quality of the acquired data. When the area of the temporary experiment is not well monitored by the local permanent network, the real-time data from the temporary experiment can be fed to the permanent network monitoring system improving greatly both the real-time hypocentral locations and the final revised bulletin. All these benefits apply also in case of seismic crises when rapid deployment stations can significantly contribute to the aftershock analysis. Nowadays data transmission using meshed radio networks or satellite systems is not a big technological problem for a permanent seismic network where each site is optimized for the device power consumption and is usually installed by properly specialized technicians that can configure transmission devices and align antennas. This is not usually practical for temporary networks and especially for rapid response networks where the installation time is the main concern. These difficulties are substantially lowered using the now widespread UMTS technology for data transmission. A small (but sometimes power hungry) properly configured device with an omnidirectional antenna must be added to the station assembly. All setups are usually configured before deployment and this allows for an easy installation also by untrained personnel. We describe here the implementation of a UMTS based portable seismic network for both temporary experiments and rapid response applications developed at INGV. The first field experimentation of this approach dates back to the 2009 L

  17. Evaluation of two real time PCR assays for the detection of bacterial DNA in amniotic fluid.

    Science.gov (United States)

    Girón de Velasco-Sada, Patricia; Falces-Romero, Iker; Quiles-Melero, Inmaculada; García-Perea, Adela; Mingorance, Jesús

    2018-01-01

    The aim of this study was to evaluate two non-commercial Real-Time PCR assays for the detection of microorganisms in amniotic fluid followed by identification by pyrosequencing. We collected 126 amniotic fluids from 2010 to 2015 for the evaluation of two Real-Time PCR assays for detection of bacterial DNA in amniotic fluid (16S Universal PCR and Ureaplasma spp. specific PCR). The method was developed in the Department of Microbiology of the University Hospital La Paz. Thirty-seven samples (29.3%) were positive by PCR/pyrosequencing and/or culture, 4 of them were mixed cultures with Ureaplasma urealyticum. The Universal 16S Real-Time PCR was compared with the standard culture (81.8% sensitivity, 97.4% specificity, 75% positive predictive value, 98% negative predictive value). The Ureaplasma spp. specific Real-Time PCR was compared with the Ureaplasma/Mycoplasma specific culture (92.3% sensitivity, 89.4% specificity, 50% positive predictive value, 99% negative predictive value) with statistically significant difference (p=0.005). Ureaplasma spp. PCR shows a rapid response time (5h from DNA extraction until pyrosequencing) when comparing with culture (48h). So, the response time of bacteriological diagnosis in suspected chorioamnionitis is reduced. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Rapid Modeling of and Response to Large Earthquakes Using Real-Time GPS Networks (Invited)

    Science.gov (United States)

    Crowell, B. W.; Bock, Y.; Squibb, M. B.

    2010-12-01

    Real-time GPS networks have the advantage of capturing motions throughout the entire earthquake cycle (interseismic, seismic, coseismic, postseismic), and because of this, are ideal for real-time monitoring of fault slip in the region. Real-time GPS networks provide the perfect supplement to seismic networks, which operate with lower noise and higher sampling rates than GPS networks, but only measure accelerations or velocities, putting them at a supreme disadvantage for ascertaining the full extent of slip during a large earthquake in real-time. Here we report on two examples of rapid modeling of recent large earthquakes near large regional real-time GPS networks. The first utilizes Japan’s GEONET consisting of about 1200 stations during the 2003 Mw 8.3 Tokachi-Oki earthquake about 100 km offshore Hokkaido Island and the second investigates the 2010 Mw 7.2 El Mayor-Cucapah earthquake recorded by more than 100 stations in the California Real Time Network. The principal components of strain were computed throughout the networks and utilized as a trigger to initiate earthquake modeling. Total displacement waveforms were then computed in a simulated real-time fashion using a real-time network adjustment algorithm that fixes a station far away from the rupture to obtain a stable reference frame. Initial peak ground displacement measurements can then be used to obtain an initial size through scaling relationships. Finally, a full coseismic model of the event can be run minutes after the event, given predefined fault geometries, allowing emergency first responders and researchers to pinpoint the regions of highest damage. Furthermore, we are also investigating using total displacement waveforms for real-time moment tensor inversions to look at spatiotemporal variations in slip.

  19. Sarcastic sentiment detection in tweets streamed in real time: a big data approach

    Directory of Open Access Journals (Sweden)

    S.K. Bharti

    2016-08-01

    Full Text Available Sarcasm is a type of sentiment where people express their negative feelings using positive or intensified positive words in the text. While speaking, people often use heavy tonal stress and certain gestural clues like rolling of the eyes, hand movement, etc. to reveal sarcastic. In the textual data, these tonal and gestural clues are missing, making sarcasm detection very difficult for an average human. Due to these challenges, researchers show interest in sarcasm detection of social media text, especially in tweets. Rapid growth of tweets in volume and its analysis pose major challenges. In this paper, we proposed a Hadoop based framework that captures real time tweets and processes it with a set of algorithms which identifies sarcastic sentiment effectively. We observe that the elapse time for analyzing and processing under Hadoop based framework significantly outperforms the conventional methods and is more suited for real time streaming tweets.

  20. Real time loss detection for SNM in process

    International Nuclear Information System (INIS)

    Candy, J.V.; Dunn, D.R.; Gavel, D.T.

    1980-01-01

    This paper discusses the basis of a design for real time special nuclear material (SNM) loss detectors. The design utilizes process measurements and signal processing techniques to produce a timely estimate of material loss. A state estimator is employed as the primary signal processing algorithm. Material loss is indicated by changes in the states or process innovations (residuals). The design philosophy is discussed in the context of these changes

  1. Real-Time RT-PCR for the Detection of Lyssavirus Species

    Directory of Open Access Journals (Sweden)

    A. Deubelbeiss

    2014-01-01

    Full Text Available The causative agents of rabies are single-stranded, negative-sense RNA viruses in the genus Lyssavirus of Rhabdoviridae, consisting of twelve classified and three as yet unclassified species including classical rabies virus (RABV. Highly neurotropic RABV causes rapidly progressive encephalomyelitis with nearly invariable fatal outcome. Rapid and reliable diagnosis of rabies is highly relevant for public and veterinary health. Due to growing variety of the genus Lyssavirus observed, the development of suitable molecular assays for diagnosis and differentiation is challenging. This work focused on the establishment of a suitable real-time RT-PCR technique for rabies diagnosis as a complement to fluorescent antibody test and rabies tissue culture infection test as gold standard for diagnosis and confirmation. The real-time RT-PCR was adapted with the goal to detect the whole spectrum of lyssavirus species, for nine of which synthesized DNA fragments were used. For the detection of species, seven probes were developed. Serial dilutions of the rabies virus strain CVS-11 showed a 100-fold higher sensitivity of real-time PCR compared to heminested RT-PCR. Using a panel of thirty-one lyssaviruses representing four species, the suitability of the protocol could be shown. Phylogenetic analysis of the sequences obtained by heminested PCR allowed correct classification of all viruses used.

  2. Detection of Tomato black ring virus by real-time one-step RT-PCR.

    Science.gov (United States)

    Harper, Scott J; Delmiglio, Catia; Ward, Lisa I; Clover, Gerard R G

    2011-01-01

    A TaqMan-based real-time one-step RT-PCR assay was developed for the rapid detection of Tomato black ring virus (TBRV), a significant plant pathogen which infects a wide range of economically important crops. Primers and a probe were designed against existing genomic sequences to amplify a 72 bp fragment from RNA-2. The assay amplified all isolates of TBRV tested, but no amplification was observed from the RNA of other nepovirus species or healthy host plants. The detection limit of the assay was estimated to be around nine copies of the TBRV target region in total RNA. A comparison with conventional RT-PCR and ELISA, indicated that ELISA, the current standard test method, lacked specificity and reacted to all nepovirus species tested, while conventional RT-PCR was approximately ten-fold less sensitive than the real-time RT-PCR assay. Finally, the real-time RT-PCR assay was tested using five different RT-PCR reagent kits and was found to be robust and reliable, with no significant differences in sensitivity being found. The development of this rapid assay should aid in quarantine and post-border surveys for regulatory agencies. Copyright © 2010 Elsevier B.V. All rights reserved.

  3. Development of a real-time multiplex PCR assay for the detection of multiple Salmonella serotypes in chicken samples

    Directory of Open Access Journals (Sweden)

    Whyte Paul

    2008-09-01

    Full Text Available Abstract Background A real-time multiplex PCR assay was developed for the detection of multiple Salmonella serotypes in chicken samples. Poultry-associated serotypes detected in the assay include Enteritidis, Gallinarum, Typhimurium, Kentucky and Dublin. The traditional cultural method according to EN ISO 6579:2002 for the detection of Salmonella in food was performed in parallel. The real-time PCR based method comprised a pre-enrichment step in Buffered Peptone Water (BPW overnight, followed by a shortened selective enrichment in Rappaport Vasilliadis Soya Broth (RVS for 6 hours and subsequent DNA extraction. Results The real-time multiplex PCR assay and traditional cultural method showed 100% inclusivity and 100% exclusivity on all strains tested. The real-time multiplex PCR assay was as sensitive as the traditional cultural method in detecting Salmonella in artificially contaminated chicken samples and correctly identified the serotype. Artificially contaminated chicken samples resulted in a detection limit of between 1 and 10 CFU per 25 g sample for both methods. A total of sixty-three naturally contaminated chicken samples were investigated by both methods and relative accuracy, relative sensitivity and relative specificity of the real-time PCR method were determined to be 89, 94 and 87%, respectively. Thirty cultures blind tested were correctly identified by the real-time multiplex PCR method. Conclusion Real-time PCR methodology can contribute to meet the need for rapid identification and detection methods in food testing laboratories.

  4. A real-time PCR assay for the detection of atypical strains of Chlamydiaceae from pigeons.

    Directory of Open Access Journals (Sweden)

    Aleksandar Zocevic

    Full Text Available Recent evidence of the occurrence of atypical Chlamydiaceae strains in pigeons, different from the established Chlamydiaceae, requires the development of a specific and rapid detection tool to investigate their prevalence and significance. Here is described a new real-time PCR assay that allows specific detection of atypical Chlamydiaceae from pigeons. The assay has been used to assess the dissemination of these strains in field samples collected from Parisian pigeon populations in 2009. The results suggest a limited dissemination compared to the usually higher prevalence of Chlamydia psittaci that is the main species associated with avian chlamydiosis.

  5. Real-Time PCR Assay To Detect Smallpox Virus

    Science.gov (United States)

    Sofi Ibrahim, M.; Kulesh, David A.; Saleh, Sharron S.; Damon, Inger K.; Esposito, Joseph J.; Schmaljohn, Alan L.; Jahrling, Peter B.

    2003-01-01

    We developed a highly sensitive and specific assay for the rapid detection of smallpox virus DNA on both the Smart Cycler and LightCycler platforms. The assay is based on TaqMan chemistry with the orthopoxvirus hemagglutinin gene used as the target sequence. With genomic DNA purified from variola virus Bangladesh 1975, the limit of detection was estimated to be approximately 25 copies on both machines. The assay was evaluated in a blinded study with 322 coded samples that included genomic DNA from 48 different isolates of variola virus; 25 different strains and isolates of camelpox, cowpox, ectromelia, gerbilpox, herpes, monkeypox, myxoma, rabbitpox, raccoonpox, skunkpox, vaccinia, and varicella-zoster viruses; and two rickettsial species at concentrations mostly ranging from 100 fg/μl to 1 ng/μl. Contained within those 322 samples were variola virus DNA, obtained from purified viral preparations, at concentrations of 1 fg/μl to 1 ng/μl. On the Smart Cycler platform, 2 samples with false-positive results were detected among the 116 samples not containing variola virus tested; i.e., the overall specificity of the assay was 98.3%. On the LightCycler platform, five samples with false-positive results were detected (overall specificity, 95.7%). Of the 206 samples that contained variola virus DNA ranging in concentrations from 100 fg/μl to 1 ng/μl, 8 samples were considered negative on the Smart Cycler platform and 1 sample was considered negative on the LightCycler platform. Thus, the clinical sensitivities were 96.1% for the Smart Cycler instrument and 99.5% for the LightCycler instrument. The vast majority of these samples were derived from virus-infected cell cultures and variola virus-infected tissues; thus, the DNA material contained both viral DNA and cellular DNA. Of the 43 samples that contained purified variola virus DNA ranging in concentration from 1 fg/μl to 1 ng/μl, the assay correctly detected the virus in all 43 samples on both the Smart Cycler

  6. A comparison of moving object detection methods for real-time moving object detection

    Science.gov (United States)

    Roshan, Aditya; Zhang, Yun

    2014-06-01

    Moving object detection has a wide variety of applications from traffic monitoring, site monitoring, automatic theft identification, face detection to military surveillance. Many methods have been developed across the globe for moving object detection, but it is very difficult to find one which can work globally in all situations and with different types of videos. The purpose of this paper is to evaluate existing moving object detection methods which can be implemented in software on a desktop or laptop, for real time object detection. There are several moving object detection methods noted in the literature, but few of them are suitable for real time moving object detection. Most of the methods which provide for real time movement are further limited by the number of objects and the scene complexity. This paper evaluates the four most commonly used moving object detection methods as background subtraction technique, Gaussian mixture model, wavelet based and optical flow based methods. The work is based on evaluation of these four moving object detection methods using two (2) different sets of cameras and two (2) different scenes. The moving object detection methods have been implemented using MatLab and results are compared based on completeness of detected objects, noise, light change sensitivity, processing time etc. After comparison, it is observed that optical flow based method took least processing time and successfully detected boundary of moving objects which also implies that it can be implemented for real-time moving object detection.

  7. Development of multiplex real-time PCR assay for the detection of Brucella spp., Leptospira spp. and Campylobacter foetus

    Directory of Open Access Journals (Sweden)

    Abdelfattah M. Selim

    2014-12-01

    Full Text Available Abortion among dairy cattle is one of the major causes of economic losses in the livestock industry. This study describes a 1-step multiplex real-time polymerase chain reaction (PCR to detect Brucella spp., Leptospira spp. and Campylobacter foetus, these are significant bacteria commonly implicated in bovine abortion. ß-actin was added to the same PCR reaction as an internal control to detect any extraction failure or PCR inhibition. The detection limit of multiplex real-time PCR using purified DNA from cultured organisms was set to 5 fg for Leptospira spp. and C. foetus and to 50 fg for Brucella spp. The multiplex real-time PCR did not produce any non-specific amplification when tested with different strains of the 3 pathogens. This multiplex real-time PCR provides a valuable tool for diagnosis, simultaneous and rapid detection for the 3 pathogens causing abortion in bovine.

  8. Development of a real-time PCR for the detection of pathogenic Leptospira spp. in California sea lions.

    Science.gov (United States)

    Wu, Qingzhong; Prager, Katherine C; Goldstein, Tracey; Alt, David P; Galloway, Renee L; Zuerner, Richard L; Lloyd-Smith, James O; Schwacke, Lori

    2014-08-11

    Several real-time PCR assays are currently used for detection of pathogenic Leptospira spp.; however, few methods have been described for the successful evaluation of clinical urine samples. This study reports a rapid assay for the detection of pathogenic Leptospira spp. in California sea lions Zalophus californianus using real-time PCR with primers and a probe targeting the lipL32 gene. The PCR assay had high analytic sensitivity-the limit of detection was 3 genome copies per PCR volume using L. interrogans serovar Pomona DNA and 100% analytic specificity; it detected all pathogenic leptospiral serovars tested and none of the non-pathogenic Leptospira species (L. biflexa and L. meyeri serovar Semaranga), the intermediate species L. inadai, or the non-Leptospira pathogens tested. Our assay had an amplification efficiency of 1.00. Comparisons between the real-time PCR assay and culture isolation for detection of pathogenic Leptospira spp. in urine and kidney tissue samples from California sea lions showed that samples were more often positive by real-time PCR than by culture methods. Inclusion of an internal amplification control in the real-time PCR assay showed no inhibitory effects in PCR negative samples. These studies indicated that our real-time PCR assay has high analytic sensitivity and specificity for the rapid detection of pathogenic Leptospira species in urine and kidney tissue samples.

  9. Rapid quantification of semen hepatitis B virus DNA by real-time polymerase chain reaction

    Science.gov (United States)

    Qian, Wei-Ping; Tan, Yue-Qiu; Chen, Ying; Peng, Ying; Li, Zhi; Lu, Guang-Xiu; Lin, Marie C.; Kung, Hsiang-Fu; He, Ming-Ling; Shing, Li-Ka

    2005-01-01

    AIM: To examine the sensitivity and accuracy of real-time polymerase chain reaction (PCR) for the quantification of hepatitis B virus (HBV) DNA in semen. METHODS: Hepatitis B viral DNA was isolated from HBV carriers’ semen and sera using phenol extraction method and QIAamp DNA blood mini kit (Qiagen, Germany). HBV DNA was detected by conventional PCR and quantified by TaqMan technology-based real-time PCR (quantitative polymerase chain reaction (qPCR)). The detection threshold was 200 copies of HBV DNA for conventional PCR and 10 copies of HBV DNA for real time PCR per reaction. RESULTS: Both methods of phenol extraction and QIAamp DNA blood mini kit were suitable for isolating HBV DNA from semen. The value of the detection thresholds was 500 copies of HBV DNA per mL in the semen. The viral loads were 7.5 × 107 and 1.67 × 107 copies of HBV DNA per mL in two HBV infected patients’ sera, while 2.14 × 105 and 3.02 × 105 copies of HBV DNA per mL in the semen. CONCLUSION: Real-time PCR is a more sensitive and accurate method to detect and quantify HBV DNA in the semen. PMID:16149152

  10. Development of a Rapid and Sensitive Method Combining a Cellulose Ester Microfilter and a Real-Time Quantitative PCR Assay To Detect Campylobacter jejuni and Campylobacter coli in 20 Liters of Drinking Water or Low-Turbidity Waters

    Science.gov (United States)

    Tissier, Adeline; Denis, Martine; Hartemann, Philippe

    2012-01-01

    Investigations of Campylobacter jejuni and Campylobacter coli in samples of drinking water suspected of being at the origin of an outbreak very often lead to negative results. One of the reasons for this failure is the small volume of water typically used for detecting these pathogens (10 to 1,000 ml). The efficiencies of three microfilters and different elution procedures were determined using real-time quantitative PCR to propose a procedure allowing detection of Campylobacter in 20 liters of drinking water or low-turbidity water samples. The results showed that more than 80% of the bacteria inoculated in 1 liter of drinking water were retained on each microfilter. An elution with a solution containing 3% beef extract, 0.05 M glycine at pH 9, combined with direct extraction of the bacterial genomes retained on the cellulose ester microfilter, allowed recovery of 87.3% (±22% [standard deviation]) of Campylobacter per 1 liter of tap water. Recoveries obtained from 20-liter volumes of tap water spiked with a C. coli strain were 69.5% (±10.3%) and 78.5% (±15.1%) for 91 CFU and 36 CFU, respectively. Finally, tests performed on eight samples of 20 liters of groundwater collected from an alluvial well used for the production of drinking water revealed the presence of C. jejuni and C. coli genomes, whereas no bacteria were detected with the normative culture method in volumes ranging from 10 to 1,000 ml. In the absence of available epidemiological data and information on bacterial viability, these last results indicate only that the water resource is not protected from contamination by Campylobacter. PMID:22138985

  11. REAL-TIME OBJECT DETECTION IN PARALLEL THROUGH ATOMIC TRANSACTIONS

    Directory of Open Access Journals (Sweden)

    K Sivakumar

    2016-11-01

    Full Text Available Object detection and tracking is important operation involved in embedded systems like video surveillance, Traffic monitoring, campus security system, machine vision applications and other areas. Detecting and tracking multiple objects in a video or image is challenging problem in machine vision and computer vision based embedded systems. Implementation of such a object detection and tracking systems are done in sequential way of processing and also it was implemented using hardware synthesize tools like verilog HDL with FPGA, achieves considerably lesser performance in speed and it does support lesser atomic transactions. There are many object detection and tracking algorithm were proposed and implemented, among them background subtraction is one of them. This paper proposes a implementation of detecting and tracking multiple objects based on background subtraction algorithm using java and .NET and also discuss about the architecture concept for object detection through atomic transactional, modern hardware synthesizes language called Bluespec.

  12. On-site identification of meat species in processed foods by a rapid real-time polymerase chain reaction system.

    Science.gov (United States)

    Furutani, Shunsuke; Hagihara, Yoshihisa; Nagai, Hidenori

    2017-09-01

    Correct labeling of foods is critical for consumers who wish to avoid a specific meat species for religious or cultural reasons. Therefore, gene-based point-of-care food analysis by real-time Polymerase Chain Reaction (PCR) is expected to contribute to the quality control in the food industry. In this study, we perform rapid identification of meat species by our portable rapid real-time PCR system, following a very simple DNA extraction method. Applying these techniques, we correctly identified beef, pork, chicken, rabbit, horse, and mutton in processed foods in 20min. Our system was sensitive enough to detect the interfusion of about 0.1% chicken egg-derived DNA in a processed food sample. Our rapid real-time PCR system is expected to contribute to the quality control in food industries because it can be applied for the identification of meat species, and future applications can expand its functionality to the detection of genetically modified organisms or mutations. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Real-time object detection, tracking and occlusion reasoning

    Science.gov (United States)

    Divakaran, Ajay; Yu, Qian; Tamrakar, Amir; Sawhney, Harpreet Singh; Zhu, Jiejie; Javed, Omar; Liu, Jingen; Cheng, Hui; Eledath, Jayakrishnan

    2018-02-27

    A system for object detection and tracking includes technologies to, among other things, detect and track moving objects, such as pedestrians and/or vehicles, in a real-world environment, handle static and dynamic occlusions, and continue tracking moving objects across the fields of view of multiple different cameras.

  14. Real-time pose invariant logo and pattern detection

    Science.gov (United States)

    Sidla, Oliver; Kottmann, Michal; Benesova, Wanda

    2011-01-01

    The detection of pose invariant planar patterns has many practical applications in computer vision and surveillance systems. The recognition of company logos is used in market studies to examine the visibility and frequency of logos in advertisement. Danger signs on vehicles could be detected to trigger warning systems in tunnels, or brand detection on transport vehicles can be used to count company-specific traffic. We present the results of a study on planar pattern detection which is based on keypoint detection and matching of distortion invariant 2d feature descriptors. Specifically we look at the keypoint detectors of type: i) Lowe's DoG approximation from the SURF algorithm, ii) the Harris Corner Detector, iii) the FAST Corner Detector and iv) Lepetit's keypoint detector. Our study then compares the feature descriptors SURF and compact signatures based on Random Ferns: we use 3 sets of sample images to detect and match 3 logos of different structure to find out which combinations of keypoint detector/feature descriptors work well. A real-world test tries to detect vehicles with a distinctive logo in an outdoor environment under realistic lighting and weather conditions: a camera was mounted on a suitable location for observing the entrance to a parking area so that incoming vehicles could be monitored. In this 2 hour long recording we can successfully detect a specific company logo without false positives.

  15. A Metrics-Based Approach to Intrusion Detection System Evaluation for Distributed Real-Time Systems

    Science.gov (United States)

    2002-04-01

    Based Approach to Intrusion Detection System Evaluation for Distributed Real - Time Systems Authors: G. A. Fink, B. L. Chappell, T. G. Turner, and...Distributed, Security. 1 Introduction Processing and cost requirements are driving future naval combat platforms to use distributed, real - time systems of...distributed, real - time systems . As these systems grow more complex, the timing requirements do not diminish; indeed, they may become more constrained

  16. Real-time incident detection using social media data.

    Science.gov (United States)

    2016-05-09

    The effectiveness of traditional incident detection is often limited by sparse sensor coverage, and reporting incidents to emergency response systems : is labor-intensive. This research project mines tweet texts to extract incident information on bot...

  17. Real time algorithms for sharp wave ripple detection.

    Science.gov (United States)

    Sethi, Ankit; Kemere, Caleb

    2014-01-01

    Neural activity during sharp wave ripples (SWR), short bursts of co-ordinated oscillatory activity in the CA1 region of the rodent hippocampus, is implicated in a variety of memory functions from consolidation to recall. Detection of these events in an algorithmic framework, has thus far relied on simple thresholding techniques with heuristically derived parameters. This study is an investigation into testing and improving the current methods for detection of SWR events in neural recordings. We propose and profile methods to reduce latency in ripple detection. Proposed algorithms are tested on simulated ripple data. The findings show that simple realtime algorithms can improve upon existing power thresholding methods and can detect ripple activity with latencies in the range of 10-20 ms.

  18. Simplified Pan-species Real-time PCR-based Detection of Plasmodium Spp. in Blood Smear.

    Science.gov (United States)

    Hassanpour, Gholamreza; Mirhendi, Hossein; Mohebali, Mehdi; Raeisi, Ahmad; Zeraati, Hojjat; Keshavarz, Hossein

    2016-01-01

    We aimed to quicken and simplify the detection of Plasmodium in blood samples by developing and testing a pan- Plasmodium real-time PCR for accurate screening of individuals suspected of malaria. A single primer/probe set for pan-species Plasmodium -specific real time PCR targeting a conserved region of the small subunit 18S ribosomal DNA was designed and evaluated for rapid diagnosis and screening of malaria infections using dried blood smears. FTA cards were used for rapid and simple DNA extraction. The primers and probes showed a positive response with the DNA extracted from bloods infected with P. falciparum and P. vivax but not with DNA extracted from various smears from uninfected blood samples. Seven positive cases positive by both microscopy and nested PCR were found among 280 blood samples taken from in South and Southeast Iran. Five samples were identified as positive for P. vivax and two as positive for P. falciparum . All positive samples were positive by real-time PCR. Furthermore, all 38-blood samples positive by microscopy were positive by real-time PCR. No microscopy-negative samples were positive by real-time PCR. By using a simple FTA card for DNA extraction and by application of the real-time PCR developed in this study, sensitivity similar to nested-PCR and microscopy was achieved. This format simplifies the detection of Plasmodium in large numbers of samples.

  19. Simplified Pan-species Real-time PCR-based Detection of Plasmodium Spp. in Blood Smear

    Directory of Open Access Journals (Sweden)

    Gholamreza HASSANPOUR

    2016-12-01

    Full Text Available Background: We aimed to quicken and simplify the detection of Plasmodium in blood samples by developing and testing a pan-Plasmodium real-time PCR for accurate screening of individuals suspected of malaria.Methods: A single primer/probe set for pan-species Plasmodium-specific real time PCR targeting a conserved region of the small subunit 18S ribosomal DNA was designed and evaluated for rapid diagnosis and screening of malaria infections using dried blood smears. FTA cards were used for rapid and simple DNA extraction.Results: The primers and probes showed a positive response with the DNA extracted from bloods infected with P. falciparum and P. vivax but not with DNA extracted from various smears from uninfected blood samples. Seven positive cases positive by both microscopy and nested PCR were found among 280 blood samples taken from in South and Southeast Iran. Five samples were identified as positive for P. vivax and two as positive for P. falciparum. All positive samples were positive by real-time PCR. Furthermore, all 38-blood samples positive by microscopy were positive by real-time PCR. No microscopy-negative samples were positive by real-time PCR.Conclusion: By using a simple FTA card for DNA extraction and by application of the real-time PCR developed in this study, sensitivity similar to nested-PCR and microscopy was achieved. This format simplifies the detection of Plasmodium in large numbers of samples.

  20. Real time risk analysis of kick detection: Testing and validation

    International Nuclear Information System (INIS)

    Islam, Rakibul; Khan, Faisal; Venkatesan, Ramchandran

    2017-01-01

    Oil and gas development is moving into harsh and remote locations where the highest level of safety is required. A blowout is one of the most feared accidents in oil and gas developments projects. The main objective of this paper is to test and validate the kick detection of blowout risk assessment model using uniquely developed experimental results. Kick detection is a major part of the blowout risk assessment model. The accuracy and timeliness of kick detection are dependent on the monitoring of multiple downhole parameters such as downhole pressure, fluid density, fluid conductivity and mass flow rate. In the present study these four parameters are considered in different logical combinations to assess the occurrence of kick and associated blowout risk. The assessed results are compared against the experimental observations. It is observed that simultaneous monitoring of mass flow rate combined with any one the three parameters provides most reliable detection of kick and potential blowout likelihood. The current work presents the framework for a dynamic risk assessment and management model. Upon success testing of this approach at the pilot and field levels, this approach could provide a paradigm shift in drilling safety. - Highlights: • A novel dynamic risk model of kick detection and blowout prediction. • Testing and Validation of the risk model. • Application of the dynamic risk model.

  1. Fast real-time polymerase chain reaction for quantitative detection of Lactobacillus delbrueckii bacteriophages in milk.

    Science.gov (United States)

    Martín, Maria Cruz; del Rio, Beatriz; Martínez, Noelia; Magadán, Alfonso H; Alvarez, Miguel A

    2008-12-01

    One of the main microbiological problems of the dairy industry is the susceptibility of starter bacteria to virus infections. Lactobacillus delbrueckii, a component of thermophilic starter cultures used in the manufacture of several fermented dairy products, including yogurt, is also sensitive to bacteriophage attacks. To avoid the problems associated with these viruses, quick and sensitive detection methods are necessary. In the present study, a fast real-time quantitative polymerase chain reaction assay for the direct detection and quantification of L. delbrueckii phages in milk was developed. A set of primers and a TaqMan MGB probe was designed, based on the lysin gene sequence of different L. delbrueckii phages. The results show the proposed method to be a rapid (total processing time 30 min), specific and highly sensitive technique for detecting L. delbrueckii phages in milk.

  2. Real-time progressive hyperspectral image processing endmember finding and anomaly detection

    CERN Document Server

    Chang, Chein-I

    2016-01-01

    The book covers the most crucial parts of real-time hyperspectral image processing: causality and real-time capability. Recently, two new concepts of real time hyperspectral image processing, Progressive Hyperspectral Imaging (PHSI) and Recursive Hyperspectral Imaging (RHSI). Both of these can be used to design algorithms and also form an integral part of real time hyperpsectral image processing. This book focuses on progressive nature in algorithms on their real-time and causal processing implementation in two major applications, endmember finding and anomaly detection, both of which are fundamental tasks in hyperspectral imaging but generally not encountered in multispectral imaging. This book is written to particularly address PHSI in real time processing, while a book, Recursive Hyperspectral Sample and Band Processing: Algorithm Architecture and Implementation (Springer 2016) can be considered as its companion book. Includes preliminary background which is essential to those who work in hyperspectral ima...

  3. Real-Time Barcode Detection and Classification Using Deep Learning

    DEFF Research Database (Denmark)

    Hansen, Daniel Kold; Nasrollahi, Kamal; Rasmussen, Christoffer Bøgelund

    2017-01-01

    Barcodes, in their different forms, can be found on almost any packages available in the market. Detecting and then decoding of barcodes have therefore great applications. We describe how to adapt the state-of-the- art deep learning-based detector of You Only Look Once (YOLO) for the purpose...

  4. Parametric Roll - Risk Reduction through Real-time Detection

    DEFF Research Database (Denmark)

    Galeazzi, Roberto

    2014-01-01

    PAROLL is an innovative condition-monitoring system for the timely detection of parametric roll on merchant vessels. It has been invented and developed by the Technical University of Denmark. DNV GL and Wallenius Marine have supported the development and full-scale validation of this monitoring...

  5. Real - time NASBA detection of strawberry vein banding virus

    Czech Academy of Sciences Publication Activity Database

    Hanzlíková-Vašková, Dana; Špak, Josef; Klerks, M. M.; Schoen, C. D.; Thompson, J. R.; Jelkmann, W.

    2004-01-01

    Roč. 110, - (2004), s. 213-221 ISSN 0929-1873 Grant - others:EU(XE) QLRT-PL99-1553 Institutional research plan: CEZ:AV0Z5051902 Keywords : strawberry virus * detection Subject RIV: EE - Microbiology, Virology Impact factor: 1.384, year: 2004

  6. Real-time Microseismic Processing for Induced Seismicity Hazard Detection

    Energy Technology Data Exchange (ETDEWEB)

    Matzel, Eric M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2016-10-31

    Induced seismicity is inherently associated with underground fluid injections. If fluids are injected in proximity to a pre-existing fault or fracture system, the resulting elevated pressures can trigger dynamic earthquake slip, which could both damage surface structures and create new migration pathways. The goal of this research is to develop a fundamentally better approach to geological site characterization and early hazard detection. We combine innovative techniques for analyzing microseismic data with a physics-based inversion model to forecast microseismic cloud evolution. The key challenge is that faults at risk of slipping are often too small to detect during the site characterization phase. Our objective is to devise fast-running methodologies that will allow field operators to respond quickly to changing subsurface conditions.

  7. Real-time PCR for detection of Theileria equi and Babesia caballi ...

    African Journals Online (AJOL)

    Real-time PCR for detection of Theileria equi and Babesia caballi parasites in ticks. ... This study aimed to develop a real-time PCR screening test for Babesia caballi and Theileria equi in ticks. Adult D. reticulatus were ... This test is suitable for application in epidemiological surveillance of equine babesiosis and theileriosis.

  8. Research on rapid agile metrology for manufacturing based on real-time multitask operating system

    Science.gov (United States)

    Chen, Jihong; Song, Zhen; Yang, Daoshan; Zhou, Ji; Buckley, Shawn

    1996-10-01

    Rapid agile metrology for manufacturing (RAMM) using multiple non-contact sensors is likely to remain a growing trend in manufacturing. High speed inspecting systems for manufacturing is characterized by multitasks implemented in parallel and real-time events which occur simultaneously. In this paper, we introduce a real-time operating system into RAMM research. A general task model of a class-based object- oriented technology is proposed. A general multitask frame of a typical RAMM system using OPNet is discussed. Finally, an application example of a machine which inspects parts held on a carrier strip is described. With RTOS and OPNet, this machine can measure two dimensions of the contacts at 300 parts/second.

  9. Real-time PCR improves Helicobacter pylori detection in patients with peptic ulcer bleeding.

    Directory of Open Access Journals (Sweden)

    María José Ramírez-Lázaro

    Full Text Available BACKGROUND AND AIMS: Histological and rapid urease tests to detect H. pylori in biopsy specimens obtained during peptic ulcer bleeding episodes (PUB often produce false-negative results. We aimed to examine whether immunohistochemistry and real-time PCR can improve the sensitivity of these biopsies. PATIENTS AND METHODS: We selected 52 histology-negative formalin-fixed paraffin-embedded biopsy specimens obtained during PUB episodes. Additional tests showed 10 were true negatives and 42 were false negatives. We also selected 17 histology-positive biopsy specimens obtained during PUB to use as controls. We performed immunohistochemistry staining and real-time PCR for 16S rRNA, ureA, and 23S rRNA for H. pylori genes on all specimens. RESULTS: All controls were positive for H. pylori on all PCR assays and immunohistochemical staining. Regarding the 52 initially negative biopsies, all PCR tests were significantly more sensitive than immunohistochemical staining (p<0.01. Sensitivity and specificity were 55% and 80% for 16S rRNA PCR, 43% and 90% for ureA PCR, 41% and 80% for 23S rRNA PCR, and 7% and 100% for immunohistochemical staining, respectively. Combined analysis of PCR assays for two genes were significantly more sensitive than ureA or 23S rRNA PCR tests alone (p<0.05 and marginally better than 16S rRNA PCR alone. The best combination was 16S rRNA+ureA, with a sensitivity of 64% and a specificity of 80%. CONCLUSIONS: Real-time PCR improves the detection of H. pylori infection in histology-negative formalin-fixed paraffin-embedded biopsy samples obtained during PUB episodes. The low reported prevalence of H. pylori in PUB may be due to the failure of conventional tests to detect infection.

  10. Real time QRS complex detection using DFA and regular grammar.

    Science.gov (United States)

    Hamdi, Salah; Ben Abdallah, Asma; Bedoui, Mohamed Hedi

    2017-02-28

    The sequence of Q, R, and S peaks (QRS) complex detection is a crucial procedure in electrocardiogram (ECG) processing and analysis. We propose a novel approach for QRS complex detection based on the deterministic finite automata with the addition of some constraints. This paper confirms that regular grammar is useful for extracting QRS complexes and interpreting normalized ECG signals. A QRS is assimilated to a pair of adjacent peaks which meet certain criteria of standard deviation and duration. The proposed method was applied on several kinds of ECG signals issued from the standard MIT-BIH arrhythmia database. A total of 48 signals were used. For an input signal, several parameters were determined, such as QRS durations, RR distances, and the peaks' amplitudes. σRR and σQRS parameters were added to quantify the regularity of RR distances and QRS durations, respectively. The sensitivity rate of the suggested method was 99.74% and the specificity rate was 99.86%. Moreover, the sensitivity and the specificity rates variations according to the Signal-to-Noise Ratio were performed. Regular grammar with the addition of some constraints and deterministic automata proved functional for ECG signals diagnosis. Compared to statistical methods, the use of grammar provides satisfactory and competitive results and indices that are comparable to or even better than those cited in the literature.

  11. Real-time defect detection on highly reflective curved surfaces

    Science.gov (United States)

    Rosati, G.; Boschetti, G.; Biondi, A.; Rossi, A.

    2009-03-01

    This paper presents an automated defect detection system for coated plastic components for the automotive industry. This research activity came up as an evolution of a previous study which employed a non-flat mirror to illuminate and inspect high reflective curved surfaces. According to this method, the rays emitted from a light source are conveyed on the surface under investigation by means of a suitably curved mirror. After the reflection on the surface, the light rays are collected by a CCD camera, in which the coating defects appear as shadows of various shapes and dimensions. In this paper we present an evolution of the above-mentioned method, introducing a simplified mirror set-up in order to reduce the costs and the complexity of the defect detection system. In fact, a set of plane mirrors is employed instead of the curved one. Moreover, the inspection of multiple bend radius parts is investigated. A prototype of the machine vision system has been developed in order to test this simplified method. This device is made up of a light projector, a set of plane mirrors for light rays reflection, a conveyor belt for handling components, a CCD camera and a desktop PC which performs image acquisition and processing. Like in the previous system, the defects are identified as shadows inside a high brightness image. At the end of the paper, first experimental results are presented.

  12. Automated Incident Detection Using Real-Time Floating Car Data

    Directory of Open Access Journals (Sweden)

    Maarten Houbraken

    2017-01-01

    Full Text Available The aim of this paper is to demonstrate the feasibility of a live Automated Incident Detection (AID system using only Floating Car Data (FCD in one of the first large-scale FCD AID field trials. AID systems detect traffic events and alert upcoming drivers to improve traffic safety without human monitoring. These automated systems traditionally rely on traffic monitoring sensors embedded in the road. FCD allows for finer spatial granularity of traffic monitoring. However, low penetration rates of FCD probe vehicles and the data latency have historically hindered FCD AID deployment. We use a live country-wide FCD system monitoring an estimated 5.93% of all vehicles. An FCD AID system is presented and compared to the installed AID system (using loop sensor data on 2 different highways in Netherlands. Our results show the FCD AID can adequately monitor changing traffic conditions and follow the AID benchmark. The presented FCD AID is integrated with the road operator systems as part of an innovation project, making this, to the best of our knowledge, the first full chain technical feasibility trial of an FCD-only AID system. Additionally, FCD allows for AID on roads without installed sensors, allowing road safety improvements at low cost.

  13. WiFi-Based Real-Time Calibration-Free Passive Human Motion Detection

    Directory of Open Access Journals (Sweden)

    Liangyi Gong

    2015-12-01

    Full Text Available With the rapid development of WLAN technology, wireless device-free passive human detection becomes a newly-developing technique and holds more potential to worldwide and ubiquitous smart applications. Recently, indoor fine-grained device-free passive human motion detection based on the PHY layer information is rapidly developed. Previous wireless device-free passive human detection systems either rely on deploying specialized systems with dense transmitter-receiver links or elaborate off-line training process, which blocks rapid deployment and weakens system robustness. In the paper, we explore to research a novel fine-grained real-time calibration-free device-free passive human motion via physical layer information, which is independent of indoor scenarios and needs no prior-calibration and normal profile. We investigate sensitivities of amplitude and phase to human motion, and discover that phase feature is more sensitive to human motion, especially to slow human motion. Aiming at lightweight and robust device-free passive human motion detection, we develop two novel and practical schemes: short-term averaged variance ratio (SVR and long-term averaged variance ratio (LVR. We realize system design with commercial WiFi devices and evaluate it in typical multipath-rich indoor scenarios. As demonstrated in the experiments, our approach can achieve a high detection rate and low false positive rate.

  14. WiFi-Based Real-Time Calibration-Free Passive Human Motion Detection.

    Science.gov (United States)

    Gong, Liangyi; Yang, Wu; Man, Dapeng; Dong, Guozhong; Yu, Miao; Lv, Jiguang

    2015-12-21

    With the rapid development of WLAN technology, wireless device-free passive human detection becomes a newly-developing technique and holds more potential to worldwide and ubiquitous smart applications. Recently, indoor fine-grained device-free passive human motion detection based on the PHY layer information is rapidly developed. Previous wireless device-free passive human detection systems either rely on deploying specialized systems with dense transmitter-receiver links or elaborate off-line training process, which blocks rapid deployment and weakens system robustness. In the paper, we explore to research a novel fine-grained real-time calibration-free device-free passive human motion via physical layer information, which is independent of indoor scenarios and needs no prior-calibration and normal profile. We investigate sensitivities of amplitude and phase to human motion, and discover that phase feature is more sensitive to human motion, especially to slow human motion. Aiming at lightweight and robust device-free passive human motion detection, we develop two novel and practical schemes: short-term averaged variance ratio (SVR) and long-term averaged variance ratio (LVR). We realize system design with commercial WiFi devices and evaluate it in typical multipath-rich indoor scenarios. As demonstrated in the experiments, our approach can achieve a high detection rate and low false positive rate.

  15. WiFi-Based Real-Time Calibration-Free Passive Human Motion Detection

    Science.gov (United States)

    Gong, Liangyi; Yang, Wu; Man, Dapeng; Dong, Guozhong; Yu, Miao; Lv, Jiguang

    2015-01-01

    With the rapid development of WLAN technology, wireless device-free passive human detection becomes a newly-developing technique and holds more potential to worldwide and ubiquitous smart applications. Recently, indoor fine-grained device-free passive human motion detection based on the PHY layer information is rapidly developed. Previous wireless device-free passive human detection systems either rely on deploying specialized systems with dense transmitter-receiver links or elaborate off-line training process, which blocks rapid deployment and weakens system robustness. In the paper, we explore to research a novel fine-grained real-time calibration-free device-free passive human motion via physical layer information, which is independent of indoor scenarios and needs no prior-calibration and normal profile. We investigate sensitivities of amplitude and phase to human motion, and discover that phase feature is more sensitive to human motion, especially to slow human motion. Aiming at lightweight and robust device-free passive human motion detection, we develop two novel and practical schemes: short-term averaged variance ratio (SVR) and long-term averaged variance ratio (LVR). We realize system design with commercial WiFi devices and evaluate it in typical multipath-rich indoor scenarios. As demonstrated in the experiments, our approach can achieve a high detection rate and low false positive rate. PMID:26703612

  16. Reverse transcriptase real-time PCR for detection and quantification of viable Campylobacter jejuni directly from poultry faecal samples

    DEFF Research Database (Denmark)

    Bui, Thanh Xuan; Wolff, Anders; Madsen, Mogens

    2012-01-01

    Campylobacter spp. is the most common cause of bacterial diarrhoea in humans worldwide. Therefore, rapid and reliable methods fordetection and quantification of this pathogen are required. In this study, we have developed a reverse transcription quantitative real-time PCR(RT-qPCR) for detection a...

  17. Detecting EUV transients in near real time with ALEXIS

    Energy Technology Data Exchange (ETDEWEB)

    Roussel-Dupre`, D.; Bloch, J.J.; Theiler, J.; Pfafman, T.; Beauchesne, B.

    1995-12-31

    The Array of Low Energy X-ray Imaging Sensors (ALEXIS) experiment consists of a mini-satellite containing six wide angle EUV/ultrasoft X-ray telescopes (Priedhorsky et al. 1989, and Bloch et al. 1994). Its scientific objective is to map out the sky in three narrow ({Delta}E/E {approx} 5%) bandpasses around 66, 71, and 93 eV. During each 50 second satellite rotation period the six telescopes, each with a 30{degrees} field, of:view and a spatial resolution of 0.25{degrees}, scan most of the antisolar hemisphere of the sky. The project is a collaborative effort between Los Alamos National Laboratory, Sandia National Laboratory, and the University of California-Berkeley Space Sciences Laboratory. It is controlled entirely from a small ground station located at Los Alamos. The mission was launched on a Pegasus Air Launched Vehicle on April 25, 1993. An incident at launch delayed our ability to properly analyze the data until November of 1994. In January of 1995, we brought on line automated software to routinely carry out the transient search. After the data is downlinked from the satellite, the software processes and transforms it into sky maps that are automatically searched for new sources. The software then sends the results of these searches by e-mail to the science team within two hours of the downlink. This system has successfully detected the Cataclysmic Variables VW Hyi, U Gem and AR UMa in outburst, and has detected at least two unidentified short duration EUV transients (Roussel-Dupre et al 1995, Roussel-Dupre 1995).

  18. Radioactive aerosol detection station for near real-time atmospheric monitoring

    International Nuclear Information System (INIS)

    Mason, L.R.; Bohner, John D.

    1997-01-01

    A radionuclide aerosol detection station has been developed to measure radioactivity in the environment. The objective is to monitor the atmosphere for anthropogenic radioactivity that could be indicative of nuclear weapons tests to verify the Comprehensive Nuclear Test Ban Treaty. Eighty stations will form the backbone of the International Monitoring System in which stations are linked to a central analysis facility called the International Data Centre. Data are transmitted to this centre in near real-time to facilitate rapid detection. Principal process of the field measurement are collection, separation, and assay. Collection of airborne radioactivity is achieved through high-volume air sampling. Aerosols separation is accomplished by high-efficiency particulate filtration. Radionuclides assay is achieved by in-situ high resolution gamma spectrometry. These modules are integrated into a unit that provides power, control, and communication support subsystems. Station operation is semi-automatic requiring only minimal human interaction. (author). 6 refs., 3 figs., 3 tabs

  19. Real-time detection of antibiotic activity by measuring nanometer-scale bacterial deformation

    Science.gov (United States)

    Iriya, Rafael; Syal, Karan; Jing, Wenwen; Mo, Manni; Yu, Hui; Haydel, Shelley E.; Wang, Shaopeng; Tao, Nongjian

    2017-12-01

    Diagnosing antibiotic-resistant bacteria currently requires sensitive detection of phenotypic changes associated with antibiotic action on bacteria. Here, we present an optical imaging-based approach to quantify bacterial membrane deformation as a phenotypic feature in real-time with a nanometer scale (˜9 nm) detection limit. Using this approach, we found two types of antibiotic-induced membrane deformations in different bacterial strains: polymyxin B induced relatively uniform spatial deformation of Escherichia coli O157:H7 cells leading to change in cellular volume and ampicillin-induced localized spatial deformation leading to the formation of bulges or protrusions on uropathogenic E. coli CFT073 cells. We anticipate that the approach will contribute to understanding of antibiotic phenotypic effects on bacteria with a potential for applications in rapid antibiotic susceptibility testing.

  20. Real-Time Fluorescence Loop Mediated Isothermal Amplification for the Detection of Acinetobacter baumannii

    Science.gov (United States)

    Wang, Qinqin; Zhou, Yanbin; Li, Shaoli; Zhuo, Chao; Xu, Siqi; Huang, Lixia; Yang, Ling; Liao, Kang

    2013-01-01

    Background Detection of Acinetobacter baumannii has been relying primarily on bacterial culture that often fails to return useful results in time. Although DNA-based assays are more sensitive than bacterial culture in detecting the pathogen, the molecular results are often inconsistent and challenged by doubts on false positives, such as those due to system- and environment-derived contaminations. In addition, these molecular tools require expensive laboratory instruments. Therefore, establishing molecular tools for field use require simpler molecular platforms. The loop-mediated isothermal amplification method is relatively simple and can be improved for better use in a routine clinical bacteriology laboratory. A simple and portable device capable of performing both the amplification and detection (by fluorescence) of LAMP in the same platform has been developed in recent years. This method is referred to as real-time loop-mediated isothermal amplification. In this study, we attempted to utilize this method for rapid detection of A. baumannii. Methodology and Significant Findings Species-specific primers were designed to test the utility of this method. Clinical samples of A. baumannii were used to determine the sensitivity and specificity of this system compared to bacterial culture and a polymerase chain reaction method. All positive samples isolated from sputum were confirmed to be the species of Acinetobacter by 16S rRNA gene sequencing. The RealAmp method was found to be simpler and allowed real-time detection of DNA amplification, and could distinguish A. baumannii from Acinetobacter calcoaceticus and Acinetobacter genomic species 3. DNA was extracted by simple boiling method. Compared to bacterial culture, the sensitivity and specificity of RealAmp in detecting A. baumannii was 98.9% and 75.0%, respectively. Conclusion The RealAmp assay only requires a single unit, and the assay positivity can be verified by visual inspection. Therefore, this assay has

  1. Rapid Large Earthquake and Run-up Characterization in Quasi Real Time

    Science.gov (United States)

    Bravo, F. J.; Riquelme, S.; Koch, P.; Cararo, S.

    2017-12-01

    Several test in quasi real time have been conducted by the rapid response group at CSN (National Seismological Center) to characterize earthquakes in Real Time. These methods are known for its robustness and realibility to create Finite Fault Models. The W-phase FFM Inversion, The Wavelet Domain FFM and The Body Wave and FFM have been implemented in real time at CSN, all these algorithms are running automatically and triggered by the W-phase Point Source Inversion. Dimensions (Large and Width ) are predefined by adopting scaling laws for earthquakes in subduction zones. We tested the last four major earthquakes occurred in Chile using this scheme: The 2010 Mw 8.8 Maule Earthquake, The 2014 Mw 8.2 Iquique Earthquake, The 2015 Mw 8.3 Illapel Earthquake and The 7.6 Melinka Earthquake. We obtain many solutions as time elapses, for each one of those we calculate the run-up using an analytical formula. Our results are in agreements with some FFM already accepted by the sicentific comunnity aswell as run-up observations in the field.

  2. Detection of Listeria monocytogenes in ready-to-eat food by Step One real-time polymerase chain reaction.

    Science.gov (United States)

    Pochop, Jaroslav; Kačániová, Miroslava; Hleba, Lukáš; Lopasovský, L'ubomír; Bobková, Alica; Zeleňáková, Lucia; Stričík, Michal

    2012-01-01

    The aim of this study was to follow contamination of ready-to-eat food with Listeria monocytogenes by using the Step One real time polymerase chain reaction (PCR). We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and MicroSEQ® Listeria monocytogenes Detection Kit for the real-time PCR performance. In 30 samples of ready-to-eat milk and meat products without incubation we detected strains of Listeria monocytogenes in five samples (swabs). Internal positive control (IPC) was positive in all samples. Our results indicated that the real-time PCR assay developed in this study could sensitively detect Listeria monocytogenes in ready-to-eat food without incubation.

  3. Real-time implementation of logo detection on open source BeagleBoard

    Science.gov (United States)

    George, M.; Kehtarnavaz, N.; Estevez, L.

    2011-03-01

    This paper presents the real-time implementation of our previously developed logo detection and tracking algorithm on the open source BeagleBoard mobile platform. This platform has an OMAP processor that incorporates an ARM Cortex processor. The algorithm combines Scale Invariant Feature Transform (SIFT) with k-means clustering, online color calibration and moment invariants to robustly detect and track logos in video. Various optimization steps that are carried out to allow the real-time execution of the algorithm on BeagleBoard are discussed. The results obtained are compared to the PC real-time implementation results.

  4. DIRADTM - a system for real time detection and identification of radioactive objects

    International Nuclear Information System (INIS)

    Guillot, L.; Reboli, A.

    2009-01-01

    The authors present the DIRAD system (DIRAD stands for Detection and Identification of Radionuclides), an automatic system for real time identification of a radioactive anomaly and its interpretation in terms of risk level. It can be adapted to different contexts: pedestrian control, parcel or luggage control, road traffic control, and so on. In case of risk detection, an alert is transmitted in real time to a supervision station along with the whole set of spectral data

  5. Real-time Detection of Antihydrogen Annihilations and Applications to Spectroscopy

    Directory of Open Access Journals (Sweden)

    Stracka Simone

    2014-04-01

    Full Text Available A detection scheme based on real-time measurement of antihydrogen annihilations during radiation injection is presented, which allows an efficient use of the trapped atoms for laser and microwave spectroscopy. The application of real-time detection of H¯$\\bar H$ annihilations to microwave spectroscopy, which yielded the first evidence of microwave induced spin-flip transitions in trapped antihydrogen [1], is reported.

  6. Multispectral fluorometric sensor for real time in-situ detection of marine petroleum spills

    International Nuclear Information System (INIS)

    Andrews, J.M.; Lieberman, S.H.

    1998-01-01

    This paper describes the development of a fluorescence based in-situ sensor system for real time monitoring and detection of petroleum hydrocarbon contaminants in the marine environment. The system consists of an array of underwater sensors deployed just below the water surface. The sensors can detect floating product (surface sheen) from below the surface as well as detect emulsified or dissolved phase petroleum in the water column. Data from each of the sensors is transmitted to a central base station computer for display, logging, and analysis. The primary intended use of the system is to protect marine facilities from accidental, petroleum discharges by providing responding authorities with immediate notification of the occurrence of a leak or spill. The detection of petroleum is based upon the fluorescence of polycyclic aromatic hydrocarbons found within petroleum derived products. The sensors utilize broadband ultraviolet excitation from a pulsed xenon lamp to generate fluorescence in contaminated sea water. The intensity of the resulting fluorescence emission is proportional to both the oil concentration in water, and/or the oil film thickness on the water surface. Multispectral fluorescence emission information is used to distinguish between several possible petroleum classes and eliminate false positive interference from non-petroleum based fluorophores such as chlorophyll. Real time qualitative identification yields an important advantage in terms of rapidly resolving questions of spill origin or in determining an appropriate response. To enable long term underwater deployment, the optical energy of the ultraviolet excitation source also serves to prevent the occurrence of biofouling on the surface of the optical window. The results of initial testing in San Diego Harbor and at the Ohmsett wave tank facility in New Jersey demonstrate the system's ability to detect petroleum products under a variety of conditions, including the presence of strong harbor

  7. Development of Capillary Loop Convective Polymerase Chain Reaction Platform with Real-Time Fluorescence Detection

    Directory of Open Access Journals (Sweden)

    Wen-Pin Chou

    2017-02-01

    Full Text Available Polymerase chain reaction (PCR has been one of the principal techniques of molecular biology and diagnosis for decades. Conventional PCR platforms, which work by rapidly heating and cooling the whole vessel, need complicated hardware designs, and cause energy waste and high cost. On the other hand, partial heating on the various locations of vessels to induce convective solution flows by buoyancy have been used for DNA amplification in recent years. In this research, we develop a new convective PCR platform, capillary loop convective polymerase chain reaction (clcPCR, which can generate one direction flow and make the PCR reaction more stable. The U-shaped loop capillaries with 1.6 mm inner diameter are designed as PCR reagent containers. The clcPCR platform utilizes one isothermal heater for heating the bottom of the loop capillary and a CCD device for detecting real-time amplifying fluorescence signals. The stable flow was generated in the U-shaped container and the amplification process could be finished in 25 min. Our experiments with different initial concentrations of DNA templates demonstrate that clcPCR can be applied for precise quantification. Multiple sample testing and real-time quantification will be achieved in future studies.

  8. Development and validation of a real-time PCR assay for the detection of anguillid herpesvirus 1.

    Science.gov (United States)

    van Beurden, S J; Voorbergen-Laarman, M A; Roozenburg, I; van Tellingen, J; Haenen, O L M; Engelsma, M Y

    2016-01-01

    Anguillid herpesvirus 1 (AngHV1) causes a haemorrhagic disease with increased mortality in wild and farmed European eel, Anguilla anguilla (L.) and Japanese eel Anguilla japonica, Temminck & Schlegel). Detection of AngHV1 is currently based on virus isolation in cell culture, antibody-based typing assays or conventional PCR. We developed, optimized and concisely validated a diagnostic TaqMan probe based real-time PCR assay for the detection of AngHV1. The primers and probe target AngHV1 open reading frame 57, encoding the capsid protease and scaffold protein. Compared to conventional PCR, the developed real-time PCR is faster, less labour-intensive and has a reduced risk of cross-contamination. The real-time PCR assay was shown to be analytically sensitive and specific and has a high repeatability, efficiency and r(2) -value. The diagnostic performance of the assay was determined by testing 10% w/v organ suspensions and virus cultures from wild and farmed European eels from the Netherlands by conventional and real-time PCR. The developed real-time PCR assay is a useful tool for the rapid and sensitive detection of AngHV1 in 10% w/v organ suspensions from wild and farmed European eels. © 2015 John Wiley & Sons Ltd.

  9. Real-time biscuit tile image segmentation method based on edge detection.

    Science.gov (United States)

    Matić, Tomislav; Aleksi, Ivan; Hocenski, Željko; Kraus, Dieter

    2018-05-01

    In this paper we propose a novel real-time Biscuit Tile Segmentation (BTS) method for images from ceramic tile production line. BTS method is based on signal change detection and contour tracing with a main goal of separating tile pixels from background in images captured on the production line. Usually, human operators are visually inspecting and classifying produced ceramic tiles. Computer vision and image processing techniques can automate visual inspection process if they fulfill real-time requirements. Important step in this process is a real-time tile pixels segmentation. BTS method is implemented for parallel execution on a GPU device to satisfy the real-time constraints of tile production line. BTS method outperforms 2D threshold-based methods, 1D edge detection methods and contour-based methods. Proposed BTS method is in use in the biscuit tile production line. Copyright © 2018 ISA. Published by Elsevier Ltd. All rights reserved.

  10. Real-time sensor failure detection by dynamic modelling of a PWR plant

    International Nuclear Information System (INIS)

    Turkcan, E.; Ciftcioglu, O.

    1992-06-01

    Signal validation and sensor failure detection is an important problem in real-time nuclear power plant (NPP) surveillance. Although conventional sensor redundancy, in a way, is a solution, identification of faulty sensor is necessary for further preventive actions to be taken. A comprehensive solution for the system so that any sensory reading is verified by its model based estimated counterpart, in real-time. Such a realization is accomplished by means of dynamic system's states estimation methodology using Kalman filter modelling technique. The method is investigated by means of real-time data of the steam generator of Borssele nuclear power plant and the method has proved to be satisfactory for real-time sensor failure detection as well as model validation verification. (author). 5 refs.; 6 figs.; 1 tab

  11. Real-time detection of musical onsets with linear prediction and sinusoidal modeling

    Science.gov (United States)

    Glover, John; Lazzarini, Victor; Timoney, Joseph

    2011-12-01

    Real-time musical note onset detection plays a vital role in many audio analysis processes, such as score following, beat detection and various sound synthesis by analysis methods. This article provides a review of some of the most commonly used techniques for real-time onset detection. We suggest ways to improve these techniques by incorporating linear prediction as well as presenting a novel algorithm for real-time onset detection using sinusoidal modelling. We provide comprehensive results for both the detection accuracy and the computational performance of all of the described techniques, evaluated using Modal, our new open source library for musical onset detection, which comes with a free database of samples with hand-labelled note onsets.

  12. FPGA-Based Real-Time Motion Detection for Automated Video Surveillance Systems

    Directory of Open Access Journals (Sweden)

    Sanjay Singh

    2016-03-01

    Full Text Available Design of automated video surveillance systems is one of the exigent missions in computer vision community because of their ability to automatically select frames of interest in incoming video streams based on motion detection. This research paper focuses on the real-time hardware implementation of a motion detection algorithm for such vision based automated surveillance systems. A dedicated VLSI architecture has been proposed and designed for clustering-based motion detection scheme. The working prototype of a complete standalone automated video surveillance system, including input camera interface, designed motion detection VLSI architecture, and output display interface, with real-time relevant motion detection capabilities, has been implemented on Xilinx ML510 (Virtex-5 FX130T FPGA platform. The prototyped system robustly detects the relevant motion in real-time in live PAL (720 × 576 resolution video streams directly coming from the camera.

  13. Improvement of a real-time RT-PCR assay for the detection of enterovirus RNA

    Directory of Open Access Journals (Sweden)

    Bruynseels Peggy

    2009-07-01

    Full Text Available Abstract We describe an improvement of an earlier reported real-time RT-PCR assay for the detection of enterovirus RNA, based on the 5' exonuclease digestion of a dual-labeled fluorogenic probe by Taq DNA polymerase. A different extraction method, real-time RT-PCR instrument and primer set were evaluated. Our data show that the optimized assay yields a higher sensitivity and reproducibility and resulted in a significant reduced hands-on time per sample.

  14. Automated real-time detection of tonic-clonic seizures using a wearable EMG device

    DEFF Research Database (Denmark)

    Beniczky, Sándor; Conradsen, Isa; Henning, Oliver

    2018-01-01

    OBJECTIVE: To determine the accuracy of automated detection of generalized tonic-clonic seizures (GTCS) using a wearable surface EMG device. METHODS: We prospectively tested the technical performance and diagnostic accuracy of real-time seizure detection using a wearable surface EMG device....... The seizure detection algorithm and the cutoff values were prespecified. A total of 71 patients, referred to long-term video-EEG monitoring, on suspicion of GTCS, were recruited in 3 centers. Seizure detection was real-time and fully automated. The reference standard was the evaluation of video-EEG recordings...

  15. UMTS rapid response real-time seismic networks: implementation and strategies at INGV

    Science.gov (United States)

    Govoni, A.; Margheriti, L.; Moretti, M.; Lauciani, V.; Sensale, G.; Bucci, A.; Criscuoli, F.

    2015-12-01

    Universal Mobile Telecommunications System (UMTS) and its evolutions are nowadays the most affordable and widespread data communication infrastructure available almost world wide. Moreover the always growing cellular phone market is pushing the development of new devices with higher performances and lower power consumption. All these characteristics make UMTS really useful for the implementation of an "easy to deploy" temporary real-time seismic station. Despite these remarkable features, there are many drawbacks that must be properly taken in account to effectively transmit the seismic data: Internet security, signal and service availability, power consumption. - Internet security: exposing seismological data services and seismic stations to the Internet is dangerous, attack prone and can lead to downtimes in the services, so we setup a dedicated Virtual Private Network (VPN) service to protect all the connected devices. - Signal and service availability: while for temporary experiment a carefull planning and an accurate site selection can minimize the problem, this is not always the case with rapid response networks. Moreover, as with any other leased line, the availability of the UMTS service during a seismic crisis is basically unpredictable. Nowadays in Italy during a major national emergency a Committee of the Italian Civil Defense ensures unified management and coordination of emergency activities. Inside it the telecom companies are committed to give support to the crisis management improving the standards in their communication networks. - Power consumption: it is at least of the order of that of the seismic station and, being related to data flow and signal quality is largely unpredictable. While the most secure option consists in adding a second independent solar power supply to the seismic station, this is not always a very convenient solution since it doubles the cost and doubles the equipment on site. We found that an acceptable trade-off is to add an

  16. Improving multi-GNSS ultra-rapid orbit determination for real-time precise point positioning

    Science.gov (United States)

    Li, Xingxing; Chen, Xinghan; Ge, Maorong; Schuh, Harald

    2018-03-01

    Currently, with the rapid development of multi-constellation Global Navigation Satellite Systems (GNSS), the real-time positioning and navigation are undergoing dramatic changes with potential for a better performance. To provide more precise and reliable ultra-rapid orbits is critical for multi-GNSS real-time positioning, especially for the three merging constellations Beidou, Galileo and QZSS which are still under construction. In this contribution, we present a five-system precise orbit determination (POD) strategy to fully exploit the GPS + GLONASS + BDS + Galileo + QZSS observations from CDDIS + IGN + BKG archives for the realization of hourly five-constellation ultra-rapid orbit update. After adopting the optimized 2-day POD solution (updated every hour), the predicted orbit accuracy can be obviously improved for all the five satellite systems in comparison to the conventional 1-day POD solution (updated every 3 h). The orbit accuracy for the BDS IGSO satellites can be improved by about 80, 45 and 50% in the radial, cross and along directions, respectively, while the corresponding accuracy improvement for the BDS MEO satellites reaches about 50, 20 and 50% in the three directions, respectively. Furthermore, the multi-GNSS real-time precise point positioning (PPP) ambiguity resolution has been performed by using the improved precise satellite orbits. Numerous results indicate that combined GPS + BDS + GLONASS + Galileo (GCRE) kinematic PPP ambiguity resolution (AR) solutions can achieve the shortest time to first fix (TTFF) and highest positioning accuracy in all coordinate components. With the addition of the BDS, GLONASS and Galileo observations to the GPS-only processing, the GCRE PPP AR solution achieves the shortest average TTFF of 11 min with 7{°} cutoff elevation, while the TTFF of GPS-only, GR, GE and GC PPP AR solution is 28, 15, 20 and 17 min, respectively. As the cutoff elevation increases, the reliability and accuracy of GPS-only PPP AR solutions

  17. Detection of Histoplasma capsulatum from clinical specimens by cycling probe-based real-time PCR and nested real-time PCR.

    Science.gov (United States)

    Muraosa, Yasunori; Toyotome, Takahito; Yahiro, Maki; Watanabe, Akira; Shikanai-Yasuda, Maria Aparecida; Kamei, Katsuhiko

    2016-05-01

    We developed new cycling probe-based real-time PCR and nested real-time PCR assays for the detection of Histoplasma capsulatum that were designed to detect the gene encoding N-acetylated α-linked acidic dipeptidase (NAALADase), which we previously identified as an H. capsulatum antigen reacting with sera from patients with histoplasmosis. Both assays specifically detected the DNAs of all H. capsulatum strains but not those of other fungi or human DNA. The limited of detection (LOD) of the real-time PCR assay was 10 DNA copies when using 10-fold serial dilutions of the standard plasmid DNA and 50 DNA copies when using human serum spiked with standard plasmid DNA. The nested real-time PCR improved the LOD to 5 DNA copies when using human serum spiked with standard plasmid DNA, which represents a 10-fold higher than that observed with the real-time PCR assay. To assess the ability of the two assays to diagnose histoplasmosis, we analyzed a small number of clinical specimens collected from five patients with histoplasmosis, such as sera (n = 4), formalin-fixed paraffin-embedded (FFPE) tissue (n = 4), and bronchoalveolar lavage fluid (BALF) (n = 1). Although clinical sensitivity of the real-time PCR assay was insufficiently sensitive (33%), the nested real-time PCR assay increased the clinical sensitivity (77%), suggesting it has a potential to be a useful method for detecting H. capsulatum DNA in clinical specimens. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae

    Directory of Open Access Journals (Sweden)

    Kirsty F. Smith

    2014-03-01

    Full Text Available The identification of toxin-producing dinoflagellates for monitoring programmes and bio-compound discovery requires considerable taxonomic expertise. It can also be difficult to morphologically differentiate toxic and non-toxic species or strains. Various molecular methods have been used for dinoflagellate identification and detection, and this study describes the development of eight real-time polymerase chain reaction (PCR assays targeting the large subunit ribosomal RNA (LSU rRNA gene of species from the genera Gymnodinium, Karenia, Karlodinium, and Takayama. Assays proved to be highly specific and sensitive, and the assay for G. catenatum was further developed for quantification in response to a bloom in Manukau Harbour, New Zealand. The assay estimated cell densities from environmental samples as low as 0.07 cells per PCR reaction, which equated to three cells per litre. This assay not only enabled conclusive species identification but also detected the presence of cells below the limit of detection for light microscopy. This study demonstrates the usefulness of real-time PCR as a sensitive and rapid molecular technique for the detection and quantification of micro-algae from environmental samples.

  19. Rapid and accurate identification by real-time PCR of biotoxin-producing dinoflagellates from the family gymnodiniaceae.

    Science.gov (United States)

    Smith, Kirsty F; de Salas, Miguel; Adamson, Janet; Rhodes, Lesley L

    2014-03-07

    The identification of toxin-producing dinoflagellates for monitoring programmes and bio-compound discovery requires considerable taxonomic expertise. It can also be difficult to morphologically differentiate toxic and non-toxic species or strains. Various molecular methods have been used for dinoflagellate identification and detection, and this study describes the development of eight real-time polymerase chain reaction (PCR) assays targeting the large subunit ribosomal RNA (LSU rRNA) gene of species from the genera Gymnodinium, Karenia, Karlodinium, and Takayama. Assays proved to be highly specific and sensitive, and the assay for G. catenatum was further developed for quantification in response to a bloom in Manukau Harbour, New Zealand. The assay estimated cell densities from environmental samples as low as 0.07 cells per PCR reaction, which equated to three cells per litre. This assay not only enabled conclusive species identification but also detected the presence of cells below the limit of detection for light microscopy. This study demonstrates the usefulness of real-time PCR as a sensitive and rapid molecular technique for the detection and quantification of micro-algae from environmental samples.

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

    Science.gov (United States)

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

    2018-03-01

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

  1. Detection of Low Molecular Weight Adulterants in Beverages by Direct Analysis in Real Time Mass Spectrometry.

    Science.gov (United States)

    Sisco, Edward; Dake, Jeffrey

    2016-04-14

    Direct Analysis in Real Time Mass Spectrometry (DART-MS) has been used to detect the presence of non-narcotic adulterants in beverages. The non-narcotic adulterants that were examined in this work incorporated a number low molecular weight alcohols, acetone, ammonium hydroxide, and sodium hypochlorite. Analysis of the adulterants was completed by pipetting 1 µL deposits onto glass microcapillaries along with an appropriate dopant species followed by introduction into the DART gas stream. It was found that detection of these compounds in the complex matrices of common beverages (soda, energy drinks, etc.) was simplified through the use of a dopant species to allow for adduct formation with the desired compound(s) of interest. Other parameters that were investigated included DART gas stream temperature, in source collision induced dissociation, ion polarity, and DART needle voltage. Sensitivities of the technique were found to range from 0.001 % volume fraction to 0.1 % volume fraction, comparable to traditional analyses completed using headspace gas chromatography mass spectrometry (HS-GC/MS). Once a method was established using aqueous solutions, , fifteen beverages were spiked with each of the nine adulterants, to simulate real world detection, and in nearly all cases the adulterant could be detected either in pure form, or complexed with the added dopant species. This technique provides a rapid way to directly analyze beverages believed to be contaminated with non-narcotic adulterants at sensitivities similar to or exceeding those of traditional confirmatory analyses.

  2. Near real time detection of deforestation in the Brazilian Amazon using MODIS imagery

    Directory of Open Access Journals (Sweden)

    Egídio Arai

    2007-06-01

    Full Text Available The objective of this paper is to provide near real time information about deforestation detection (DETER in the entire Brazilian Amazon using MODIS high temporal resolution images. It is part of the operational deforestation monitoring project to estimate the annual deforestation rate in the Brazilian Amazon (PRODES. A rapid deforestation detection method was designed to support land use policies in this region. In order to evaluate the proposed method a test site was selected covering a Landsat ETM+ scene (227/68 located in Mato Grosso State. For this purpose a multitemporal series of MODIS surface reflectance images (MOD09 and the corresponding ETM+ images from June to October 2002 were analyzed. It was found that small deforested areas (lower than 15 ha were detected by MODIS images with lower accuracy when compared with ETM+ images. As the deforested areas increase MODIS and ETM+ results tend to converge. This procedure showed to be adequate to operationally detect and monitor deforested areas and has been used since 2004 as part of a government plan to control the Amazon deforestation.

  3. Feathered Detectives: Real-Time GPS Tracking of Scavenging Gulls Pinpoints Illegal Waste Dumping.

    Directory of Open Access Journals (Sweden)

    Joan Navarro

    Full Text Available Urban waste impacts human and environmental health, and waste management has become one of the major challenges of humanity. Concurrently with new directives due to manage this human by-product, illegal dumping has become one of the most lucrative activities of organized crime. Beyond economic fraud, illegal waste disposal strongly enhances uncontrolled dissemination of human pathogens, pollutants and invasive species. Here, we demonstrate the potential of novel real-time GPS tracking of scavenging species to detect environmental crime. Specifically, we were able to detect illegal activities at an officially closed dump, which was visited recurrently by 5 of 19 GPS-tracked yellow-legged gulls (Larus michahellis. In comparison with conventional land-based surveys, GPS tracking allows a much wider and cost-efficient spatiotemporal coverage, even of the most hazardous sites, while GPS data accessibility through the internet enables rapid intervention. Our results suggest that multi-species guilds of feathered detectives equipped with GPS and cameras could help fight illegal dumping at continental scales. We encourage further experimental studies, to infer waste detection thresholds in gulls and other scavenging species exploiting human waste dumps.

  4. Real time RT-PCR assay for detection of different serotypes of FMDV in Egypt

    Directory of Open Access Journals (Sweden)

    Laila El-Shehawy

    Full Text Available Aim: The present study indicated that rRT-PCR could be provided for the detection of FMDV in infected, contact and carrier cattle and also provide a rapid sensitive tool aiming to aid in rapid disease detection and control. Foot and Mouth disease virus serotypes O and A still existing in Egypt. In January 2012, sever outbreaks struck the animal population in most Egyptian 1 governorates. The causative virus was identified as FMDV SAT2. Material and Methods: Five samples of tongue epithelium (ET and five oesophageal-pharyngeal (OP fluid samples were collected from FMD suspected cattle in infected farm at El-Fayoum and 20 OP samples from in-contact cattle at the same farm in addition to 30 OP samples from apparently healthy cattle at three different localities in El-Fayoum governorate (12 from Fayoum; 9 from Sinoras and 9 from Edsa in order to detect carrier cattle. All of these samples were collected during November and December 2011 and January 2012. Results: All the ET and OP samples were inoculated on BHK cell culture and baby mice. The obtained results were identified using complement fixation test in addition to real-time reverse transcriptase polymerase chain reaction (rRT-PCR. In the infected farm at El-Fayoum FMDV type SAT2 was detected in cattle which are considered as the first introduction of this type while FMDV type O and SAT2 were detected in the in-contact cattle in the same farm. The sensitivity of rRT-PCR was cleared in the in-contact cattle as 13 out of 20 OP samples were positive to FMDV by rRT-PCR while 11 out of 20 OP samples were positive to FMDV by CFT. The OP samples collected from apparently healthy cattle from Fayoum, Sinoras and Edsa localities in Fayoum governorate demonstrate the circulation of the FMDV type A, O and the recent SAT2 in carrier cattle which threaten cattle population in Fayoum governorate. Also the sensitivity of real time RT-PCR over the CFT in detection of FMDV carrier cattle was clearly noticed in

  5. Status of IGS Ultra-Rapid Products for Real-Time Applications

    Science.gov (United States)

    Ray, J.; Griffiths, J.

    2008-12-01

    Since November 2000 the International GNSS Service (IGS) has produced Ultra-rapid (IGU) products for near real-time and real-time applications. They include GPS orbits, satellite clocks, and Earth rotation parameters for a sliding 48-hr period. The first day of each update is based on the most recent GPS observational data from the IGS hourly tracking network. At the time of release, these observed products have an initial latency of 3 hr. The second day of each update consists of predictions. So the predictions between about 3 and 9 hr into the second half are relevant for true real-time uses. Originally updated twice daily, the IGU products since April 2004 have been issued four times per day, at 3, 9, 15, and 21 UTC. Up to seven Analysis Centers (ACs) contribute to the IGU combinations: Astronomical Institute of the University of Berne (AIUB), European Space Operations Center (ESOC), Geodetic Observatory Pecny (GOP), GeoForschungsZentrum (GFZ) Potsdam, Natural Resources Canada (NRC), Scripps Insitution of Oceanography (SIO), U.S. Naval Observatory (USNO). This redundancy affords a high measure of reliability and enhanced orbit accuracy. IGU orbit precision has improved markedly since late 2007. This is due to a combination of factors: decommissioning of the old, poorly behaved PRN29 in October 2007; upgraded procedures implemented by GOP around the same time, by SIO in spring 2008, and by USNO in June 2008; better handling of maneuvered satellites at the combination level starting June 2008; and stricter AC rejection criteria since July 2008. As a consequence, the weighted 1D RMS residual of the IGU orbit predictions over their first 6 hr is currently about 20 to 30 mm (after a Helmert transformation) compared to the IGS Rapid orbits, averaged over the constellation. The median residual is about 15 to 20 mm. When extended to the full 24 hr prediction period, the IGU orbit errors approximately double. Systematic rotational offsets are probably more important than

  6. Exploring inter-frame correlation analysis and wavelet-domain modeling for real-time caption detection in streaming video

    Science.gov (United States)

    Li, Jia; Tian, Yonghong; Gao, Wen

    2008-01-01

    In recent years, the amount of streaming video has grown rapidly on the Web. Often, retrieving these streaming videos offers the challenge of indexing and analyzing the media in real time because the streams must be treated as effectively infinite in length, thus precluding offline processing. Generally speaking, captions are important semantic clues for video indexing and retrieval. However, existing caption detection methods often have difficulties to make real-time detection for streaming video, and few of them concern on the differentiation of captions from scene texts and scrolling texts. In general, these texts have different roles in streaming video retrieval. To overcome these difficulties, this paper proposes a novel approach which explores the inter-frame correlation analysis and wavelet-domain modeling for real-time caption detection in streaming video. In our approach, the inter-frame correlation information is used to distinguish caption texts from scene texts and scrolling texts. Moreover, wavelet-domain Generalized Gaussian Models (GGMs) are utilized to automatically remove non-text regions from each frame and only keep caption regions for further processing. Experiment results show that our approach is able to offer real-time caption detection with high recall and low false alarm rate, and also can effectively discern caption texts from the other texts even in low resolutions.

  7. Soil Baiting, Rapid PCR Assay and Quantitative Real Time PCR to Diagnose Late Blight of Potato in Quarantine Programs

    Directory of Open Access Journals (Sweden)

    Touseef Hussain

    2018-05-01

    Full Text Available Phytophthora infestans (mont de Bary is a pathogen of great concern across the globe, and accurate detection is an important component in responding to the outbreaks of potential disease. Although the molecular diagnostic protocol used in regulatory programs has been evaluated but till date methods implying direct comparison has rarely used. In this study, a known area soil samples from potato fields where light blight appear every year (both A1 and A2 mating type was assayed by soil bait method, PCR assay detection and quantification of the inoculums. Suspected disease symptoms appeared on bait tubers were further confirmed by rapid PCR, inoculums were quantified through Real Time PCR, which confirms presence of P. infestans. These diagnostic methods can be highly correlated with one another. Potato tuber baiting increased the sensitivity of the assay compared with direct extraction of DNA from tuber and soil samples. Our study determines diagnostic sensitivity and specificity of the assays to determine the performance of each method. Overall, molecular techniques based on different types of PCR amplification and Real-time PCR can lead to high throughput, faster and more accurate detection method which can be used in quarantine programmes in potato industry and diagnostic laboratory.

  8. Detection of three porcine vesicular viruses using multiplex real-time primer-probe energy transfer

    DEFF Research Database (Denmark)

    Rasmussen, Thomas Bruun; Uttenthal, Åse; Aguero, M.

    2006-01-01

    Rapid identification of the etiologic agent in infected animals is important for the control of an outbreak of vesicular disease in livestock. We have in the present study developed a multiplex real-time reverse transcription-PCR, based on primer-probe energy transfer (PriProET), for simultaneous...

  9. Molecular detection of Plasmodium knowlesi in a Dutch traveler by real-time PCR.

    Science.gov (United States)

    Link, Lonneke; Bart, Aldert; Verhaar, Nienke; van Gool, Tom; Pronk, Marjolijn; Scharnhorst, Volkher

    2012-07-01

    Plasmodium knowlesi infection with low parasitemia presents a diagnostic challenge, as rapid diagnostic tests are often negative and identification to the species level by microscopy is difficult. P. knowlesi malaria in a traveler is described, and real-time PCR is demonstrated to support fast and reliable diagnosis and identification to the species level.

  10. Research on conditional characteristics vision real-time detection system for conveyor belt longitudinal tear

    NARCIS (Netherlands)

    Qiao, Tiezhu; Li, Xinyu; Pang, Y.; Lü, Yuxiang; Wang, Feng; Jin, Baoquan

    2017-01-01

    Conveyor belt longitudinal tear is one of the most serious problems in coal mining. Existing systems cannot realise lossless and real-time detection for longitudinal tear of conveyor belt. Currently, visual detecting systems are proposed by many researchers and are becoming the future trend. A

  11. Real-time DDoS attack detection for Cisco IOS using NetFlow

    NARCIS (Netherlands)

    van der Steeg, Daniël; Hofstede, R.J.; Sperotto, Anna; Pras, Aiko

    Flow-based DDoS attack detection is typically performed by analysis applications that are installed on or close to a flow collector. Although this approach allows for easy deployment, it makes detection far from real-time and susceptible to DDoS attacks for the following reasons. First, the fact

  12. Detection of Campylobacter spp. in chicken fecal samples by real-time PCR

    DEFF Research Database (Denmark)

    Lund, Marianne; Nordentoft, Steen; Pedersen, Karl

    2004-01-01

    A real-time PCR assay for detecting thermophilic Campylobacter spp. directly in chicken feces has been developed. DNA was isolated from fecal material by using magnetic beads followed by PCR with a prealiquoted PCR mixture, which had been stored at -18degreesC. Campylobacter could be detected...

  13. Real-time pedestrian detection with the videos of car camera

    Directory of Open Access Journals (Sweden)

    Yunling Zhang

    2015-12-01

    Full Text Available Pedestrians in the vehicle path are in danger of being hit, thus causing severe injury to pedestrians and vehicle occupants. Therefore, real-time pedestrian detection with the video of vehicle-mounted camera is of great significance to vehicle–pedestrian collision warning and traffic safety of self-driving car. In this article, a real-time scheme was proposed based on integral channel features and graphics processing unit. The proposed method does not need to resize the input image. Moreover, the computationally expensive convolution of the detectors and the input image was converted into the dot product of two larger matrixes, which can be computed effectively using a graphics processing unit. The experiments showed that the proposed method could be employed to detect pedestrians in the video of car camera at 20+ frames per second with acceptable error rates. Thus, it can be applied in real-time detection tasks with the videos of car camera.

  14. Comparison between digital PCR and real-time PCR in detection of Salmonella typhimurium in milk.

    Science.gov (United States)

    Wang, Meng; Yang, Junjie; Gai, Zhongtao; Huo, Shengnan; Zhu, Jianhua; Li, Jun; Wang, Ranran; Xing, Sheng; Shi, Guosheng; Shi, Feng; Zhang, Lei

    2018-02-02

    As a kind of zero-tolerance foodborne pathogens, Salmonella typhimurium poses a great threat to quality of food products and public health. Hence, rapid and efficient approaches to identify Salmonella typhimurium are urgently needed. Combined with PCR and fluorescence technique, real-time PCR (qPCR) and digital PCR (ddPCR) are regarded as suitable tools for detecting foodborne pathogens. To compare the effect between qPCR and ddPCR in detecting Salmonella typhimurium, a series of nucleic acid, pure strain culture and spiking milk samples were applied and the resistance to inhibitors referred in this article as well. Compared with qPCR, ddPCR exhibited more sensitive (10 -4 ng/μl or 10 2 cfu/ml) and less pre-culturing time (saving 2h). Moreover, ddPCR had stronger resistance to inhibitors than qPCR, yet absolute quantification hardly performed when target's concentration over 1ng/μl or 10 6 cfu/ml. This study provides an alternative strategy in detecting foodborne Salmonella typhimurium. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Real-time 3-dimensional contrast-enhanced ultrasound in detecting hemorrhage of blunt renal trauma.

    Science.gov (United States)

    Xu, Rui-Xue; Li, Ye-Kuo; Li, Ting; Wang, Sha-Sha; Yuan, Gui-Zhong; Zhou, Qun-Fang; Zheng, Hai-Rong; Yan, Fei

    2013-10-01

    The objective of this study is to evaluate the diagnostic value of real-time 3-dimensional contrast-enhanced ultrasound in the hemorrhage of blunt renal trauma. Eighteen healthy New Zealand white rabbits were randomly divided into 3 groups. Blunt renal trauma was performed on each group by using minitype striker. Ultrasonography, color Doppler flow imaging, and contrast-enhanced 2-dimensional and real-time 3-dimensional ultrasound were applied before and after the strike. The time to shock and blood pressure were subjected to statistical analysis. Then, a comparative study of ultrasound and pathology was carried out. All the struck kidneys were traumatic. In the ultrasonography, free fluid was found under the renal capsule. In the color Doppler flow imaging, active hemorrhage was not identified. In 2-dimensional contrast-enhanced ultrasound, active hemorrhage of the damaged kidney was characterized. Real-time 3-dimensional contrast-enhanced ultrasound showed a real-time and stereoscopic ongoing bleeding of the injured kidney. The wider the hemorrhage area in 4-dimensional contrast-enhanced ultrasound was, the faster the blood pressure decreased. Real-time 3-dimensional contrast-enhanced ultrasound is a promising noninvasive tool for stereoscopically and vividly detecting ongoing hemorrhage of blunt renal trauma in real time. © 2013.

  16. Rapid determination of ginkgolic acids in Ginkgo biloba kernels and leaves by direct analysis in real time-mass spectrometry.

    Science.gov (United States)

    Huang, Zhongping; Xu, Yueting; Huang, Yilei; Liu, Charles; Jiang, Kezhi; Wang, Lili

    2017-12-01

    A novel method based on direct analysis in real time integrated with mass spectrometry was established and applied into rapid determination of ginkgolic acids in Ginkgo biloba kernels and leaves. Instrument parameter settings were optimized to obtain the sensitive and accurate determination of ginkgolic acids. At the sample introduction speed of 0.2 mm/s, high intensity of [M-H] - ions for ginkgolic acids were observed in the negative ion mode by utilization of high-purity helium gas at 450°C. Two microliters of methanol extract of G. biloba kernels or leaves dropped on the surface of Quick-Strip module was analyzed after solvent evaporated to dryness. A series of standard solutions of ginkgolic acid 13:0 in the range of 2-50 mg/L were analyzed with a correlation coefficient r = 0.9981 and relative standard deviation (n = 5) from 12.5 to 13.7%. The limit of detection was 0.5 mg/L. The results of direct analysis in real time-mass spectrometry were in agreement with those observed by thermochemolysis gas chromatography. The proposed method demonstrated significant potential in the application of the high-throughput screening and rapid analysis for ginkgolic acids in dietary supplements. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Real-Time GPS Monitoring for Earthquake Rapid Assessment in the San Francisco Bay Area

    Science.gov (United States)

    Guillemot, C.; Langbein, J. O.; Murray, J. R.

    2012-12-01

    The U.S. Geological Survey Earthquake Science Center has deployed a network of eight real-time Global Positioning System (GPS) stations in the San Francisco Bay area and is implementing software applications to continuously evaluate the status of the deformation within the network. Real-time monitoring of the station positions is expected to provide valuable information for rapidly estimating source parameters should a large earthquake occur in the San Francisco Bay area. Because earthquake response applications require robust data access, as a first step we have developed a suite of web-based applications which are now routinely used to monitor the network's operational status and data streaming performance. The web tools provide continuously updated displays of important telemetry parameters such as data latency and receive rates, as well as source voltage and temperature information within each instrument enclosure. Automated software on the backend uses the streaming performance data to mitigate the impact of outages, radio interference and bandwidth congestion on deformation monitoring operations. A separate set of software applications manages the recovery of lost data due to faulty communication links. Displacement estimates are computed in real-time for various combinations of USGS, Plate Boundary Observatory (PBO) and Bay Area Regional Deformation (BARD) network stations. We are currently comparing results from two software packages (one commercial and one open-source) used to process 1-Hz data on the fly and produce estimates of differential positions. The continuous monitoring of telemetry makes it possible to tune the network to minimize the impact of transient interruptions of the data flow, from one or more stations, on the estimated positions. Ongoing work is focused on using data streaming performance history to optimize the quality of the position, reduce drift and outliers by switching to the best set of stations within the network, and

  18. Ultrasensitive microchip based on smart microgel for real-time online detection of trace threat analytes.

    Science.gov (United States)

    Lin, Shuo; Wang, Wei; Ju, Xiao-Jie; Xie, Rui; Liu, Zhuang; Yu, Hai-Rong; Zhang, Chuan; Chu, Liang-Yin

    2016-02-23

    Real-time online detection of trace threat analytes is critical for global sustainability, whereas the key challenge is how to efficiently convert and amplify analyte signals into simple readouts. Here we report an ultrasensitive microfluidic platform incorporated with smart microgel for real-time online detection of trace threat analytes. The microgel can swell responding to specific stimulus in flowing solution, resulting in efficient conversion of the stimulus signal into significantly amplified signal of flow-rate change; thus highly sensitive, fast, and selective detection can be achieved. We demonstrate this by incorporating ion-recognizable microgel for detecting trace Pb(2+), and connecting our platform with pipelines of tap water and wastewater for real-time online Pb(2+) detection to achieve timely pollution warning and terminating. This work provides a generalizable platform for incorporating myriad stimuli-responsive microgels to achieve ever-better performance for real-time online detection of various trace threat molecules, and may expand the scope of applications of detection techniques.

  19. A real-time PCR assay for detection and quantification of Verticillium dahliae in spinach seed.

    Science.gov (United States)

    Duressa, Dechassa; Rauscher, Gilda; Koike, Steven T; Mou, Beiquan; Hayes, Ryan J; Maruthachalam, Karunakaran; Subbarao, Krishna V; Klosterman, Steven J

    2012-04-01

    Verticillium dahliae is a soilborne fungus that causes Verticillium wilt on multiple crops in central coastal California. Although spinach crops grown in this region for fresh and processing commercial production do not display Verticillium wilt symptoms, spinach seeds produced in the United States or Europe are commonly infected with V. dahliae. Planting of the infected seed increases the soil inoculum density and may introduce exotic strains that contribute to Verticillium wilt epidemics on lettuce and other crops grown in rotation with spinach. A sensitive, rapid, and reliable method for quantification of V. dahliae in spinach seed may help identify highly infected lots, curtail their planting, and minimize the spread of exotic strains via spinach seed. In this study, a quantitative real-time polymerase chain reaction (qPCR) assay was optimized and employed for detection and quantification of V. dahliae in spinach germplasm and 15 commercial spinach seed lots. The assay used a previously reported V. dahliae-specific primer pair (VertBt-F and VertBt-R) and an analytical mill for grinding tough spinach seed for DNA extraction. The assay enabled reliable quantification of V. dahliae in spinach seed, with a sensitivity limit of ≈1 infected seed per 100 (1.3% infection in a seed lot). The quantification was highly reproducible between replicate samples of a seed lot and in different real-time PCR instruments. When tested on commercial seed lots, a pathogen DNA content corresponding to a quantification cycle value of ≥31 corresponded with a percent seed infection of ≤1.3%. The assay is useful in qualitatively assessing seed lots for V. dahliae infection levels, and the results of the assay can be helpful to guide decisions on whether to apply seed treatments.

  20. Detection and differentiation of Mycobacterium tuberculosis and nontuberculous mycobacterial isolates by real-time PCR.

    Science.gov (United States)

    Shrestha, Nabin K; Tuohy, Marion J; Hall, Gerri S; Reischl, Udo; Gordon, Steven M; Procop, Gary W

    2003-11-01

    Mycobacteria cause a variety of illnesses that differ in severity and public health implications. The differentiation of Mycobacterium tuberculosis from nontuberculous mycobacteria (NTM) is of primary importance for infection control and choice of antimicrobial therapy. Despite advances in molecular diagnostics, the ability to rapidly diagnose M. tuberculosis infections by PCR is still inadequate, largely because of the possibility of false-negative reactions. We designed and validated a real-time PCR for mycobacteria by using the LightCycler system with 18 reference strains and 168 clinical mycobacterial isolates. All clinically significant mycobacteria were detected; the mean melting temperatures (with 99.9% confidence intervals [99.9% CI] in parentheses) for the different mycobacteria were as follows: M. tuberculosis, 64.35 degrees C (63.27 to 65.42 degrees C); M. kansasii, 59.20 degrees C (58.07 to 60.33 degrees C); M. avium, 57.82 degrees C (57.05 to 58.60 degrees C); M. intracellulare, 54.46 degrees C (53.69 to 55.23 degrees C); M. marinum, 58.91 degrees C (58.28 to 59.55 degrees C); rapidly growing mycobacteria, 53.09 degrees C (50.97 to 55.20 degrees C) or 43.19 degrees C (42.19 to 44.49 degrees C). This real-time PCR assay with melting curve analysis consistently accurately detected and differentiated M. tuberculosis from NTM. Detection of an NTM helps ensure that the negative result for M. tuberculosis is a true negative. The specific melting temperature also provides a suggestion of the identity of the NTM present, when the most commonly encountered mycobacterial species are considered. In a parallel comparison, both the LightCycler assay and the COBAS Amplicor M. tuberculosis assay correctly categorized 48 of 50 specimens that were proven by culture to contain M. tuberculosis, and the LightCycler assay correctly characterized 3 of 3 specimens that contained NTM.

  1. Automated Detection of Short Optical Transients of Astrophysical Origin in Real Time

    Directory of Open Access Journals (Sweden)

    Marcin Sokołowski

    2010-01-01

    Full Text Available The detection of short optical transients of astrophysical origin in real time is an important task for existing robotic telescopes. The faster a new optical transient is detected, the earlier follow-up observations can be started. The sooner the object is identified, the more data can be collected before the source fades away, particularly in the most interesting early period of the transient. In this the real-time pipeline designed for identification of optical flashes with the “Pi of the Sky” project will be presented in detail together with solutions used by other experiments.

  2. Sensitive detection of African swine fever virus using real-time PCR with a 5' conjugated minor groove binding probe

    DEFF Research Database (Denmark)

    McKillan, John; McMenamy, Michael; Hjertner, Bernt

    2010-01-01

    sensitive than the conventional PCR recommended by the OIE. Linear range was ten logs from 2 × 101 to 2 × 1010. The assay is rapid with an amplification time just over 2 h. The development of this assay provides a useful tool for the specific diagnosis of ASF in statutory or emergency testing programs......The design of a 5′ conjugated minor groove binder (MGB) probe real-time PCR assay is described for the rapid, sensitive and specific detection of African swine fever virus (ASFV) DNA. The assay is designed against the 9GL region and is capable of detecting 20 copies of a DNA standard. It does...

  3. On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System.

    Science.gov (United States)

    DeShields, Joseph B; Bomberger, Rachel A; Woodhall, James W; Wheeler, David L; Moroz, Natalia; Johnson, Dennis A; Tanaka, Kiwamu

    2018-02-23

    On-site diagnosis of plant diseases can be a useful tool for growers for timely decisions enabling the earlier implementation of disease management strategies that reduce the impact of the disease. Presently in many diagnostic laboratories, the polymerase chain reaction (PCR), particularly real-time PCR, is considered the most sensitive and accurate method for plant pathogen detection. However, laboratory-based PCRs typically require expensive laboratory equipment and skilled personnel. In this study, soil-borne pathogens of potato are used to demonstrate the potential for on-site molecular detection. This was achieved using a rapid and simple protocol comprising of magnetic bead-based nucleic acid extraction, portable real-time PCR (fluorogenic probe-based assay). The portable real-time PCR approach compared favorably with a laboratory-based system, detecting as few as 100 copies of DNA from Spongospora subterranea. The portable real-time PCR method developed here can serve as an alternative to laboratory-based approaches and a useful on-site tool for pathogen diagnosis.

  4. Real-Time PCR Typing of Escherichia coli Based on Multiple Single Nucleotide Polymorphisms--a Convenient and Rapid Method.

    Science.gov (United States)

    Lager, Malin; Mernelius, Sara; Löfgren, Sture; Söderman, Jan

    2016-01-01

    Healthcare-associated infections caused by Escherichia coli and antibiotic resistance due to extended-spectrum beta-lactamase (ESBL) production constitute a threat against patient safety. To identify, track, and control outbreaks and to detect emerging virulent clones, typing tools of sufficient discriminatory power that generate reproducible and unambiguous data are needed. A probe based real-time PCR method targeting multiple single nucleotide polymorphisms (SNP) was developed. The method was based on the multi locus sequence typing scheme of Institute Pasteur and by adaptation of previously described typing assays. An 8 SNP-panel that reached a Simpson's diversity index of 0.95 was established, based on analysis of sporadic E. coli cases (ESBL n = 27 and non-ESBL n = 53). This multi-SNP assay was used to identify the sequence type 131 (ST131) complex according to the Achtman's multi locus sequence typing scheme. However, it did not fully discriminate within the complex but provided a diagnostic signature that outperformed a previously described detection assay. Pulsed-field gel electrophoresis typing of isolates from a presumed outbreak (n = 22) identified two outbreaks (ST127 and ST131) and three different non-outbreak-related isolates. Multi-SNP typing generated congruent data except for one non-outbreak-related ST131 isolate. We consider multi-SNP real-time PCR typing an accessible primary generic E. coli typing tool for rapid and uniform type identification.

  5. Ubiquitous health monitoring and real-time cardiac arrhythmias detection: a case study.

    Science.gov (United States)

    Li, Jian; Zhou, Haiying; Zuo, Decheng; Hou, Kun-Mean; De Vaulx, Christophe

    2014-01-01

    As the symptoms and signs of heart diseases that cause sudden cardiac death, cardiac arrhythmia has attracted great attention. Due to limitations in time and space, traditional approaches to cardiac arrhythmias detection fail to provide a real-time continuous monitoring and testing service applicable in different environmental conditions. Integrated with the latest technologies in ECG (electrocardiograph) analysis and medical care, the pervasive computing technology makes possible the ubiquitous cardiac care services, and thus brings about new technical challenges, especially in the formation of cardiac care architecture and realization of the real-time automatic ECG detection algorithm dedicated to care devices. In this paper, a ubiquitous cardiac care prototype system is presented with its architecture framework well elaborated. This prototype system has been tested and evaluated in all the clinical-/home-/outdoor-care modes with a satisfactory performance in providing real-time continuous cardiac arrhythmias monitoring service unlimitedly adaptable in time and space.

  6. Real-time detection with AdaBoost-svm combination in various face orientation

    Science.gov (United States)

    Fhonna, R. P.; Nasution, M. K. M.; Tulus

    2018-03-01

    Most of the research has used algorithm AdaBoost-SVM for face detection. However, to our knowledge so far there is no research has been facing detection on real-time data with various orientations using the combination of AdaBoost and Support Vector Machine (SVM). Characteristics of complex and diverse face variations and real-time data in various orientations, and with a very complex application will slow down the performance of the face detection system this becomes a challenge in this research. Face orientation performed on the detection system, that is 900, 450, 00, -450, and -900. This combination method is expected to be an effective and efficient solution in various face orientations. The results showed that the highest average detection rate is on the face detection oriented 00 and the lowest detection rate is in the face orientation 900.

  7. NEAR REAL-TIME AUTOMATIC MARINE VESSEL DETECTION ON OPTICAL SATELLITE IMAGES

    Directory of Open Access Journals (Sweden)

    G. Máttyus

    2013-05-01

    Full Text Available Vessel monitoring and surveillance is important for maritime safety and security, environment protection and border control. Ship monitoring systems based on Synthetic-aperture Radar (SAR satellite images are operational. On SAR images the ships made of metal with sharp edges appear as bright dots and edges, therefore they can be well distinguished from the water. Since the radar is independent from the sun light and can acquire images also by cloudy weather and rain, it provides a reliable service. Vessel detection from spaceborne optical images (VDSOI can extend the SAR based systems by providing more frequent revisit times and overcoming some drawbacks of the SAR images (e.g. lower spatial resolution, difficult human interpretation. Optical satellite images (OSI can have a higher spatial resolution thus enabling the detection of smaller vessels and enhancing the vessel type classification. The human interpretation of an optical image is also easier than as of SAR image. In this paper I present a rapid automatic vessel detection method which uses pattern recognition methods, originally developed in the computer vision field. In the first step I train a binary classifier from image samples of vessels and background. The classifier uses simple features which can be calculated very fast. For the detection the classifier is slided along the image in various directions and scales. The detector has a cascade structure which rejects most of the background in the early stages which leads to faster execution. The detections are grouped together to avoid multiple detections. Finally the position, size(i.e. length and width and heading of the vessels is extracted from the contours of the vessel. The presented method is parallelized, thus it runs fast (in minutes for 16000 × 16000 pixels image on a multicore computer, enabling near real-time applications, e.g. one hour from image acquisition to end user.

  8. Near Real-Time Automatic Marine Vessel Detection on Optical Satellite Images

    Science.gov (United States)

    Máttyus, G.

    2013-05-01

    Vessel monitoring and surveillance is important for maritime safety and security, environment protection and border control. Ship monitoring systems based on Synthetic-aperture Radar (SAR) satellite images are operational. On SAR images the ships made of metal with sharp edges appear as bright dots and edges, therefore they can be well distinguished from the water. Since the radar is independent from the sun light and can acquire images also by cloudy weather and rain, it provides a reliable service. Vessel detection from spaceborne optical images (VDSOI) can extend the SAR based systems by providing more frequent revisit times and overcoming some drawbacks of the SAR images (e.g. lower spatial resolution, difficult human interpretation). Optical satellite images (OSI) can have a higher spatial resolution thus enabling the detection of smaller vessels and enhancing the vessel type classification. The human interpretation of an optical image is also easier than as of SAR image. In this paper I present a rapid automatic vessel detection method which uses pattern recognition methods, originally developed in the computer vision field. In the first step I train a binary classifier from image samples of vessels and background. The classifier uses simple features which can be calculated very fast. For the detection the classifier is slided along the image in various directions and scales. The detector has a cascade structure which rejects most of the background in the early stages which leads to faster execution. The detections are grouped together to avoid multiple detections. Finally the position, size(i.e. length and width) and heading of the vessels is extracted from the contours of the vessel. The presented method is parallelized, thus it runs fast (in minutes for 16000 × 16000 pixels image) on a multicore computer, enabling near real-time applications, e.g. one hour from image acquisition to end user.

  9. Clinical evaluation of β-tubulin real-time PCR for rapid diagnosis of dermatophytosis, a comparison with mycological methods.

    Science.gov (United States)

    Motamedi, Marjan; Mirhendi, Hossein; Zomorodian, Kamiar; Khodadadi, Hossein; Kharazi, Mahboobeh; Ghasemi, Zeinab; Shidfar, Mohammad Reza; Makimura, Koichi

    2017-10-01

    Following our previous report on evaluation of the beta tubulin real-time PCR for detection of dermatophytosis, this study aimed to compare the real-time PCR assay with conventional methods for the clinical assessment of its diagnostic performance. Samples from a total of 853 patients with suspected dermatophyte lesions were subjected to direct examination (all samples), culture (499 samples) and real-time PCR (all samples). Fungal DNA was extracted directly from clinical samples using a conical steel bullet, followed by purification with a commercial kit and subjected to the Taq-Man probe-based real-time PCR. The study showed that among the 499 specimens for which all three methods were used, 156 (31.2%), 128 (25.6%) and 205 (41.0%) were found to be positive by direct microscopy, culture and real-time PCR respectively. Real-time PCR significantly increased the detection rate of dermatophytes compared with microscopy (288 vs 229) with 87% concordance between the two methods. The sensitivity, specificity, positive predictive value, and negative predictive value of the real-time PCR was 87.5%, 85%, 66.5% and 95.2% respectively. Although real-time PCR performed better on skin than on nail samples, it should not yet fully replace conventional diagnosis. © 2017 Blackwell Verlag GmbH.

  10. Detection of Salmonella spp. in veterinary samples by combining selective enrichment and real-time PCR.

    Science.gov (United States)

    Goodman, Laura B; McDonough, Patrick L; Anderson, Renee R; Franklin-Guild, Rebecca J; Ryan, James R; Perkins, Gillian A; Thachil, Anil J; Glaser, Amy L; Thompson, Belinda S

    2017-11-01

    Rapid screening for enteric bacterial pathogens in clinical environments is essential for biosecurity. Salmonella found in veterinary hospitals, particularly Salmonella enterica serovar Dublin, can pose unique challenges for culture and testing because of its poor growth. Multiple Salmonella serovars including Dublin are emerging threats to public health given increasing prevalence and antimicrobial resistance. We adapted an automated food testing method to veterinary samples and evaluated the performance of the method in a variety of matrices including environmental samples ( n = 81), tissues ( n = 52), feces ( n = 148), and feed ( n = 29). A commercial kit was chosen as the basis for this approach in view of extensive performance characterizations published by multiple independent organizations. A workflow was established for efficiently and accurately testing veterinary matrices and environmental samples by use of real-time PCR after selective enrichment in Rappaport-Vassiliadis soya (RVS) medium. Using this method, the detection limit for S. Dublin improved by 100-fold over subculture on selective agars (eosin-methylene blue, brilliant green, and xylose-lysine-deoxycholate). Overall, the procedure was effective in detecting Salmonella spp. and provided next-day results.

  11. Microdroplet sandwich real-time rt-PCR for detection of pandemic and seasonal influenza subtypes.

    Directory of Open Access Journals (Sweden)

    Stephanie L Angione

    Full Text Available As demonstrated by the recent 2012/2013 flu epidemic, the continual emergence of new viral strains highlights the need for accurate medical diagnostics in multiple community settings. If rapid, robust, and sensitive diagnostics for influenza subtyping were available, it would help identify epidemics, facilitate appropriate antiviral usage, decrease inappropriate antibiotic usage, and eliminate the extra cost of unnecessary laboratory testing and treatment. Here, we describe a droplet sandwich platform that can detect influenza subtypes using real-time reverse-transcription polymerase chain reaction (rtRT-PCR. Using clinical samples collected during the 2010/11 season, we effectively differentiate between H1N1p (swine pandemic, H1N1s (seasonal, and H3N2 with an overall assay sensitivity was 96%, with 100% specificity for each subtype. Additionally, we demonstrate the ability to detect viral loads as low as 10(4 copies/mL, which is two orders of magnitude lower than viral loads in typical infected patients. This platform performs diagnostics in a miniaturized format without sacrificing any sensitivity, and can thus be easily developed into devices which are ideal for small clinics and pharmacies.

  12. [Quantitative fluorogenic real-time PCR assay for respiratory syncytial virus detection].

    Science.gov (United States)

    Zhang, Qi-wei; You, Shang-you; Sun, Ji-min; Wu, Qi; Yu, Chun-hua; Zhang, Chu-yu

    2005-07-01

    To Establish a rapid and objective quantitative fluorogenic real-time PCR assay for early detection of human respiratory syncytial virus (hRSV). Two pairs of primers and one TaqMan Fluorogenic probe that are specific for the recognition of the most conservative N gene of hRSV for virus detection with LighCycler PCR in 93 nasopharyngeal secretion specimens collected from infants and young children. The assay was compared with virus isolation, routine PCR, nested PCR, and enzyme-linked immunosorbent assay (ELISA). This TaqMan assay had a sensitivity of 1 x 10(2) cDNA copies/microl with a dynamic range between 1 x 10(2) and 1 x 10(7) cDNA copies/microl, which was the same as that of nested PCR, but 10 times more sensitive than routine PCR. The specificity of the assay was evaluated by comparing hRSV with polivirus type 1, coxsackie virus type 2, influenza A, influenza B and adenovirus type 7. A PCR product of the expected size (195 bp) was produced and fluorescence signal detected for hRSV, but not for any of the other viruses. The results in LightCycler and Rotor-Gene instrument were consistent. Forty-four specimens (43.9%) were hRSV-positive with this assay and 4 (4/93,4.3%) were hRSV-positive with ELISA, showing rather low correlation between the two methods. No visible relation was found between the concentration of hRSV RNA and severity of the disease. This assay is rapid, sensitive, specific and quantitative, and has the potential of wide application for early diagnosis of hRSV infection and evaluation of the therapeutic effect.

  13. Detecting changes in real-time data: a user's guide to optimal detection.

    Science.gov (United States)

    Johnson, P; Moriarty, J; Peskir, G

    2017-08-13

    The real-time detection of changes in a noisily observed signal is an important problem in applied science and engineering. The study of parametric optimal detection theory began in the 1930s, motivated by applications in production and defence. Today this theory, which aims to minimize a given measure of detection delay under accuracy constraints, finds applications in domains including radar, sonar, seismic activity, global positioning, psychological testing, quality control, communications and power systems engineering. This paper reviews developments in optimal detection theory and sequential analysis, including sequential hypothesis testing and change-point detection, in both Bayesian and classical (non-Bayesian) settings. For clarity of exposition, we work in discrete time and provide a brief discussion of the continuous time setting, including recent developments using stochastic calculus. Different measures of detection delay are presented, together with the corresponding optimal solutions. We emphasize the important role of the signal-to-noise ratio and discuss both the underlying assumptions and some typical applications for each formulation.This article is part of the themed issue 'Energy management: flexibility, risk and optimization'. © 2017 The Author(s).

  14. Video-based real-time on-street parking occupancy detection system

    Science.gov (United States)

    Bulan, Orhan; Loce, Robert P.; Wu, Wencheng; Wang, YaoRong; Bernal, Edgar A.; Fan, Zhigang

    2013-10-01

    Urban parking management is receiving significant attention due to its potential to reduce traffic congestion, fuel consumption, and emissions. Real-time parking occupancy detection is a critical component of on-street parking management systems, where occupancy information is relayed to drivers via smart phone apps, radio, Internet, on-road signs, or global positioning system auxiliary signals. Video-based parking occupancy detection systems can provide a cost-effective solution to the sensing task while providing additional functionality for traffic law enforcement and surveillance. We present a video-based on-street parking occupancy detection system that can operate in real time. Our system accounts for the inherent challenges that exist in on-street parking settings, including illumination changes, rain, shadows, occlusions, and camera motion. Our method utilizes several components from video processing and computer vision for motion detection, background subtraction, and vehicle detection. We also present three traffic law enforcement applications: parking angle violation detection, parking boundary violation detection, and exclusion zone violation detection, which can be integrated into the parking occupancy cameras as a value-added option. Our experimental results show that the proposed parking occupancy detection method performs in real-time at 5 frames/s and achieves better than 90% detection accuracy across several days of videos captured in a busy street block under various weather conditions such as sunny, cloudy, and rainy, among others.

  15. Rapid screening of β-Globin gene mutations by Real-Time PCR in ...

    African Journals Online (AJOL)

    Introduction of the real time PCR has made a revolution in the time taken for the PCR reactions. We present a method for the diagnosis of the common mutations of the B-thalassemia in Egyptian children & families. The procedure depends on the real-time PCR using specific fluorescently labeled hybridization probes.

  16. ICARES: a real-time automated detection tool for clusters of infectious diseases in the Netherlands.

    NARCIS (Netherlands)

    Groeneveld, Geert H; Dalhuijsen, Anton; Kara-Zaïtri, Chakib; Hamilton, Bob; de Waal, Margot W; van Dissel, Jaap T; van Steenbergen, Jim E

    2017-01-01

    Clusters of infectious diseases are frequently detected late. Real-time, detailed information about an evolving cluster and possible associated conditions is essential for local policy makers, travelers planning to visit the area, and the local population. This is currently illustrated in the Zika

  17. Real-time underwater object detection based on an electrically scanned high-resolution sonar

    DEFF Research Database (Denmark)

    Henriksen, Lars

    1994-01-01

    The paper describes an approach to real time detection and tracking of underwater objects, using image sequences from an electrically scanned high-resolution sonar. The use of a high resolution sonar provides a good estimate of the location of the objects, but strains the computers on board, beca...

  18. A Real-Time Plagiarism Detection Tool for Computer-Based Assessments

    Science.gov (United States)

    Jeske, Heimo J.; Lall, Manoj; Kogeda, Okuthe P.

    2018-01-01

    Aim/Purpose: The aim of this article is to develop a tool to detect plagiarism in real time amongst students being evaluated for learning in a computer-based assessment setting. Background: Cheating or copying all or part of source code of a program is a serious concern to academic institutions. Many academic institutions apply a combination of…

  19. Real-time PCR detection of Brucella spp. DNA in lesions and viscera of bovine carcasses.

    Science.gov (United States)

    Sola, Marília Cristina; da Veiga Jardim, Eurione A G; de Freitas, Marcius Ribeiro; de Mesquita, Albenones José

    2014-09-01

    This study reports a real-time PCR assay for the detection of Brucella spp. associated with the FTA® Elute method in lesions observed during sanitary inspections in beef slaughter. Of the total 276 samples, 78 (28.3%) tested positive and 198 (71.7%) negative for Brucella spp. The real-time PCR technique associated with the FTA® Elute method proved to be an important tool for the diagnosis, judgment about and disposal of carcasses and viscera of slaughtered animals. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Rapid quality assessment of Radix Aconiti Preparata using direct analysis in real time mass spectrometry.

    Science.gov (United States)

    Zhu, Hongbin; Wang, Chunyan; Qi, Yao; Song, Fengrui; Liu, Zhiqiang; Liu, Shuying

    2012-11-08

    This study presents a novel and rapid method to identify chemical markers for the quality control of Radix Aconiti Preparata, a world widely used traditional herbal medicine. In the method, the samples with a fast extraction procedure were analyzed using direct analysis in real time mass spectrometry (DART MS) combined with multivariate data analysis. At present, the quality assessment approach of Radix Aconiti Preparata was based on the two processing methods recorded in Chinese Pharmacopoeia for the purpose of reducing the toxicity of Radix Aconiti and ensuring its clinical therapeutic efficacy. In order to ensure the safety and effectivity in clinical use, the processing degree of Radix Aconiti should be well controlled and assessed. In the paper, hierarchical cluster analysis and principal component analysis were performed to evaluate the DART MS data of Radix Aconiti Preparata samples in different processing times. The results showed that the well processed Radix Aconiti Preparata, unqualified processed and the raw Radix Aconiti could be clustered reasonably corresponding to their constituents. The loading plot shows that the main chemical markers having the most influence on the discrimination amongst the qualified and unqualified samples were mainly some monoester diterpenoid aconitines and diester diterpenoid aconitines, i.e. benzoylmesaconine, hypaconitine, mesaconitine, neoline, benzoylhypaconine, benzoylaconine, fuziline, aconitine and 10-OH-mesaconitine. The established DART MS approach in combination with multivariate data analysis provides a very flexible and reliable method for quality assessment of toxic herbal medicine. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. The development and application of the two real-time RT-PCR assays to detect the pathogen of HFMD.

    Directory of Open Access Journals (Sweden)

    Aili Cui

    Full Text Available Large-scale Hand, Foot, and Mouth Disease (HFMD outbreaks have frequently occurred in China since 2008, affecting more than one million children and causing several hundred children deaths every year. The pathogens of HFMD are mainly human enteroviruses (HEVs. Among them, human enterovirus 71 (HEV71 and coxsackievirus A16 (CVA16 are the most common pathogens of HFMD. However, other HEVs could also cause HFMD. To rapidly detect HEV71 and CVA16, and ensure detection of all HEVs causing HFMD, two real-time hybridization probe-based RT-PCR assays were developed in this study. One is a multiplex real-time RT-PCR assay, which was developed to detect and differentiate HEV71 specifically from CVA16 directly from clinical specimens within 1-2 h, and the other is a broad-spectrum real-time RT-PCR assay, which targeted almost all HEVs. The experiments confirmed that the two assays have high sensitivity and specificity, and the sensitivity was up to 0.1 TCID50/ml for detection of HEVs, HEV71, and CVA16, respectively. A total of 213 clinical specimens were simultaneously detected by three kinds of assays, including the two real-time RT-PCR assays, direct conventional RT-PCR assay, and virus isolation assay on human rhabdomyosarcoma cells (RD cells. The total positive rate of both HEV71 and CVA16 was 69.48% with real-time RT-PCR assay, 47.42% with RT-PCR assay, and 34.58% with virus isolation assay. One HFMD clinical specimen was positive for HEV, but negative for HEV71 or CVA16, which was identified as Echovirus 11 (Echo11 by virus isolation, RT-PCR, and sequencing for the VP1 gene. The two real-time RT-PCR assays had been applied in 31 provincial HFMD labs to detect the pathogens of HFMD, which has contributed to the rapid identification of the pathogens in the early stages of HFMD outbreaks, and helped to clarify the etiologic agents of HFMD in China.

  2. Performances of the New Real Time Tsunami Detection Algorithm applied to tide gauges data

    Science.gov (United States)

    Chierici, F.; Embriaco, D.; Morucci, S.

    2017-12-01

    Real-time tsunami detection algorithms play a key role in any Tsunami Early Warning System. We have developed a new algorithm for tsunami detection (TDA) based on the real-time tide removal and real-time band-pass filtering of seabed pressure time series acquired by Bottom Pressure Recorders. The TDA algorithm greatly increases the tsunami detection probability, shortens the detection delay and enhances detection reliability with respect to the most widely used tsunami detection algorithm, while containing the computational cost. The algorithm is designed to be used also in autonomous early warning systems with a set of input parameters and procedures which can be reconfigured in real time. We have also developed a methodology based on Monte Carlo simulations to test the tsunami detection algorithms. The algorithm performance is estimated by defining and evaluating statistical parameters, namely the detection probability, the detection delay, which are functions of the tsunami amplitude and wavelength, and the occurring rate of false alarms. In this work we present the performance of the TDA algorithm applied to tide gauge data. We have adapted the new tsunami detection algorithm and the Monte Carlo test methodology to tide gauges. Sea level data acquired by coastal tide gauges in different locations and environmental conditions have been used in order to consider real working scenarios in the test. We also present an application of the algorithm to the tsunami event generated by Tohoku earthquake on March 11th 2011, using data recorded by several tide gauges scattered all over the Pacific area.

  3. European validation of Real-Time PCR method for detection of Salmonella spp. in pork meat.

    Science.gov (United States)

    Delibato, Elisabetta; Rodriguez-Lazaro, David; Gianfranceschi, Monica; De Cesare, Alessandra; Comin, Damiano; Gattuso, Antonietta; Hernandez, Marta; Sonnessa, Michele; Pasquali, Frédérique; Sreter-Lancz, Zuzsanna; Saiz-Abajo, María-José; Pérez-De-Juan, Javier; Butrón, Javier; Prukner-Radovcic, Estella; Horvatek Tomic, Danijela; Johannessen, Gro S; Jakočiūnė, Džiuginta; Olsen, John E; Chemaly, Marianne; Le Gall, Francoise; González-García, Patricia; Lettini, Antonia Anna; Lukac, Maja; Quesne, Segolénè; Zampieron, Claudia; De Santis, Paola; Lovari, Sarah; Bertasi, Barbara; Pavoni, Enrico; Proroga, Yolande T R; Capuano, Federico; Manfreda, Gerardo; De Medici, Dario

    2014-08-01

    The classical microbiological method for detection of Salmonella spp. requires more than five days for final confirmation, and consequently there is a need for an alternative methodology for detection of this pathogen particularly in those food categories with a short shelf-life. This study presents an international (at European level) ISO 16140-based validation study of a non-proprietary Real-Time PCR-based method that can generate final results the day following sample analysis. It is based on an ISO compatible enrichment coupled to an easy and inexpensive DNA extraction and a consolidated Real-Time PCR assay. Thirteen laboratories from seven European Countries participated to this trial, and pork meat was selected as food model. The limit of detection observed was down to 10 CFU per 25 g of sample, showing excellent concordance and accordance values between samples and laboratories (100%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (100%) when the results obtained for the Real-Time PCR-based methods were compared to those of the ISO 6579:2002 standard method. The results of this international trial demonstrate that the evaluated Real-Time PCR-based method represents an excellent alternative to the ISO standard. In fact, it shows an equal and solid performance as well as it reduces dramatically the extent of the analytical process, and can be easily implemented routinely by the Competent Authorities and Food Industry laboratories. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Rapid quality assessment of Radix Aconiti Preparata using direct analysis in real time mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Zhu Hongbin; Wang Chunyan; Qi Yao [Changchun Center of Mass Spectrometry and Chemical Biology Laboratory, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022 (China); University of Chinese Academy of Sciences, Beijing 100039 (China); Song Fengrui, E-mail: songfr@ciac.jl.cn [Changchun Center of Mass Spectrometry and Chemical Biology Laboratory, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022 (China); Liu Zhiqiang; Liu Shuying [Changchun Center of Mass Spectrometry and Chemical Biology Laboratory, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022 (China)

    2012-11-08

    Highlights: Black-Right-Pointing-Pointer DART MS combined with PCA and HCA was used to rapidly identify markers of Radix Aconiti. Black-Right-Pointing-Pointer The DART MS behavior of six aconitine-type alkaloids was investigated. Black-Right-Pointing-Pointer Chemical markers were recognized between the qualified and unqualified samples. Black-Right-Pointing-Pointer DART MS was shown to be an effective tool for quality control of Radix Aconiti Preparata. - Abstract: This study presents a novel and rapid method to identify chemical markers for the quality control of Radix Aconiti Preparata, a world widely used traditional herbal medicine. In the method, the samples with a fast extraction procedure were analyzed using direct analysis in real time mass spectrometry (DART MS) combined with multivariate data analysis. At present, the quality assessment approach of Radix Aconiti Preparata was based on the two processing methods recorded in Chinese Pharmacopoeia for the purpose of reducing the toxicity of Radix Aconiti and ensuring its clinical therapeutic efficacy. In order to ensure the safety and effectivity in clinical use, the processing degree of Radix Aconiti should be well controlled and assessed. In the paper, hierarchical cluster analysis and principal component analysis were performed to evaluate the DART MS data of Radix Aconiti Preparata samples in different processing times. The results showed that the well processed Radix Aconiti Preparata, unqualified processed and the raw Radix Aconiti could be clustered reasonably corresponding to their constituents. The loading plot shows that the main chemical markers having the most influence on the discrimination amongst the qualified and unqualified samples were mainly some monoester diterpenoid aconitines and diester diterpenoid aconitines, i.e. benzoylmesaconine, hypaconitine, mesaconitine, neoline, benzoylhypaconine, benzoylaconine, fuziline, aconitine and 10-OH-mesaconitine. The established DART MS approach in

  5. Rapid quality assessment of Radix Aconiti Preparata using direct analysis in real time mass spectrometry

    International Nuclear Information System (INIS)

    Zhu Hongbin; Wang Chunyan; Qi Yao; Song Fengrui; Liu Zhiqiang; Liu Shuying

    2012-01-01

    Highlights: ► DART MS combined with PCA and HCA was used to rapidly identify markers of Radix Aconiti. ► The DART MS behavior of six aconitine-type alkaloids was investigated. ► Chemical markers were recognized between the qualified and unqualified samples. ► DART MS was shown to be an effective tool for quality control of Radix Aconiti Preparata. - Abstract: This study presents a novel and rapid method to identify chemical markers for the quality control of Radix Aconiti Preparata, a world widely used traditional herbal medicine. In the method, the samples with a fast extraction procedure were analyzed using direct analysis in real time mass spectrometry (DART MS) combined with multivariate data analysis. At present, the quality assessment approach of Radix Aconiti Preparata was based on the two processing methods recorded in Chinese Pharmacopoeia for the purpose of reducing the toxicity of Radix Aconiti and ensuring its clinical therapeutic efficacy. In order to ensure the safety and effectivity in clinical use, the processing degree of Radix Aconiti should be well controlled and assessed. In the paper, hierarchical cluster analysis and principal component analysis were performed to evaluate the DART MS data of Radix Aconiti Preparata samples in different processing times. The results showed that the well processed Radix Aconiti Preparata, unqualified processed and the raw Radix Aconiti could be clustered reasonably corresponding to their constituents. The loading plot shows that the main chemical markers having the most influence on the discrimination amongst the qualified and unqualified samples were mainly some monoester diterpenoid aconitines and diester diterpenoid aconitines, i.e. benzoylmesaconine, hypaconitine, mesaconitine, neoline, benzoylhypaconine, benzoylaconine, fuziline, aconitine and 10-OH-mesaconitine. The established DART MS approach in combination with multivariate data analysis provides a very flexible and reliable method for quality

  6. Effect of a Real-Time Electronic Dashboard on a Rapid Response System.

    Science.gov (United States)

    Fletcher, Grant S; Aaronson, Barry A; White, Andrew A; Julka, Reena

    2017-11-20

    A rapid response system (RRS) may have limited effectiveness when inpatient providers fail to recognize signs of early patient decompensation. We evaluated the impact of an electronic medical record (EMR)-based alerting dashboard on outcomes associated with RRS activation. We used a repeated treatment study in which the dashboard display was successively turned on and off each week for ten 2-week cycles over a 20-week period on the inpatient acute care wards of an academic medical center. The Rapid Response Team (RRT) dashboard displayed all hospital patients in a single view ranked by severity score, updated in real time. The dashboard could be seen within the EMR by any provider, including RRT members. The primary outcomes were the incidence rate ratio (IRR) of all RRT activations, unexpected ICU transfers, cardiopulmonary arrests and deaths on general medical-surgical wards (wards). We conducted an exploratory analysis of first RRT activations. There were 6736 eligible admissions during the 20-week study period. There was no change in overall RRT activations (IRR = 1.14, p = 0.07), but a significant increase in first RRT activations (IRR = 1.20, p = 0.04). There were no significant differences in unexpected ICU transfers (IRR = 1.15, p = 0.25), cardiopulmonary arrests on general wards (IRR = 1.46, p = 0.43), or deaths on general wards (IRR = 0.96, p = 0.89). The introduction of the RRT dashboard was associated with increased initial RRT activations but not overall activations, unexpected ICU transfers, cardiopulmonary arrests, or death. The RRT dashboard is a novel tool to help providers recognize patient decompensation and may improve initial RRT notification.

  7. Optimization of a Real Time PCR based method for the detection of Listeria monocytogenes in pork meat.

    Science.gov (United States)

    Gattuso, Antonietta; Gianfranceschi, Monica Virginia; Sonnessa, Michele; Delibato, Elisabetta; Marchesan, Massimo; Hernandez, Marta; De Medici, Dario; Rodriguez-Lazaro, David

    2014-08-01

    The aim of this study was to optimize a Real-Time PCR protocol for a rapid detection of Listeria monocytogenes in pork meat, using reduced volumes of primary selective enrichment broth and times of incubation to decrease the cost and time for analysis. Forty-five samples of pork meat were artificially contaminated with two different levels of L. monocytogenes (1-10 CFU per sample and 10-100 CFU per sample), homogenized in three different volumes of Half Fraser Broth (1:3; 1:5 and 1:10) and incubated at 30°C ± 1°C for 5h, 8h and 24h. The detection was conducted in parallel by Real-Time PCR and the ISO standard 11290-1 methods. L. monocytogenes was detected in all the samples after 24h by Real-Time PCR method, also using reduced volumes of Half Fraser Broth. This represents a clear advantage as the time to final detection and the inherent costs were significantly reduced compared to the ISO reference method. All samples artificially contaminated were correctly detected also after 8 of incubation at 30°C ± 1°C in Half Fraser Broth and 24h in Fraser Broth at 37°C ± 1°C using cultural method. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Real-time vehicle detection and tracking in video based on faster R-CNN

    Science.gov (United States)

    Zhang, Yongjie; Wang, Jian; Yang, Xin

    2017-08-01

    Vehicle detection and tracking is a significant part in auxiliary vehicle driving system. Using the traditional detection method based on image information has encountered enormous difficulties, especially in complex background. To solve this problem, a detection method based on deep learning, Faster R-CNN, which has very high detection accuracy and flexibility, is introduced. An algorithm of target tracking with the combination of Camshift and Kalman filter is proposed for vehicle tracking. The computation time of Faster R-CNN cannot achieve realtime detection. We use multi-thread technique to detect and track vehicle by parallel computation for real-time application.

  9. Rapid diagnostic tests as a source of DNA for Plasmodium species-specific real-time PCR

    Directory of Open Access Journals (Sweden)

    Van Esbroeck Marjan

    2011-03-01

    Full Text Available Abstract Background This study describes the use of malaria rapid diagnostic tests (RDTs as a source of DNA for Plasmodium species-specific real-time PCR. Methods First, the best method to recover DNA from RDTs was investigated and then the applicability of this DNA extraction method was assessed on 12 different RDT brands. Finally, two RDT brands (OptiMAL Rapid Malaria Test and SDFK60 malaria Ag Plasmodium falciparum/Pan test were comprehensively evaluated on a panel of clinical samples submitted for routine malaria diagnosis at ITM. DNA amplification was done with the 18S rRNA real-time PCR targeting the four Plasmodium species. Results of PCR on RDT were compared to those obtained by PCR on whole blood samples. Results Best results were obtained by isolating DNA from the proximal part of the nitrocellulose component of the RDT strip with a simple DNA elution method. The PCR on RDT showed a detection limit of 0.02 asexual parasites/μl, which was identical to the same PCR on whole blood. For all 12 RDT brands tested, DNA was detected except for one brand when a low parasite density sample was applied. In RDTs with a plastic seal covering the nitrocellulose strip, DNA extraction was hampered. PCR analysis on clinical RDT samples demonstrated correct identification for single species infections for all RDT samples with asexual parasites of P. falciparum (n = 60, Plasmodium vivax (n = 10, Plasmodium ovale (n = 10 and Plasmodium malariae (n = 10. Samples with only gametocytes were detected in all OptiMAL and in 10 of the 11 SDFK60 tests. None of the negative samples (n = 20 gave a signal by PCR on RDT. With PCR on RDT, higher Ct-values were observed than with PCR on whole blood, with a mean difference of 2.68 for OptiMAL and 3.53 for SDFK60. Mixed infections were correctly identified with PCR on RDT in 4/5 OptiMAL tests and 2/5 SDFK60 tests. Conclusions RDTs are a reliable source of DNA for Plasmodium real-time PCR. This study demonstrates the

  10. Detection of Ophiocordyceps sinensis in soil by quantitative real-time PCR.

    Science.gov (United States)

    Peng, Qingyun; Zhong, Xin; Lei, Wei; Zhang, Guren; Liu, Xin

    2013-03-01

    Ophiocordyceps sinensis, one of the best known entomopathogenic fungi in traditional Chinese medicine, parasitizes larvae of the moth genus Thitarodes, which lives in soil tunnels. However, little is known about the spatial distribution of O. sinensis in the soil. We established a protocol for DNA extraction, purification, and quantification of O. sinensis in soil with quantitative real-time PCR targeting the internal transcribed spacer region. The method was assessed using 34 soil samples from Tibet. No inhibitory effects in purified soil DNA extracts were detected. The standard curve method for absolute DNA quantification generated crossing point values that were strongly and linearly correlated to the log10 of the initial amount of O. sinensis genomic DNA (r(2) = 0.999) over 7 orders of magnitude (4 × 10(1) to 4 × 10(7) fg). The amplification efficiency and y-intercept value of the standard curve were 1.953 and 37.70, respectively. The amount of O. sinensis genomic DNA decreased with increasing soil depth and horizontal distance from a sclerotium (P protocol is rapid, specific, sensitive, and provides a powerful tool for quantification of O. sinensis from soil.

  11. Real-time detection and characterization of nuclear explosion using broadband analyses of regional seismic stations

    Science.gov (United States)

    Prastowo, T.; Madlazim

    2018-01-01

    This preliminary study aims to propose a new method of real-time detection and characterization of nuclear explosions by analyzing broadband seismic waveforms acquired from a network of regional seismic stations. Signal identification generated by a nuclear test was differentiated from natural sources of either earthquakes or other natural seismo-tectonic events by verifying crucial parameters, namely source depth, type of first motion, and P-wave domination of the broadband seismic wavesunder consideration. We examined and analyzed a recently hypothetical nuclear test performed by the North Koreangovernment that occurred on September 3, 2017 as a vital point to study. From spectral analyses, we found that the source of corresponding signals associated with detonations of the latest underground nuclear test was at a much shallower depth below the surface relatively compared with that of natural earthquakes, the suspected nuclear explosions produced compressional waves with radially directed outward from the source for their first motions, and the waves were only dominated by P-components. The results are then discussed in the context of potential uses of the proposed methodology for human-induced disaster early warning system and/or the need of rapid response purposes for minimizing the disaster risks.

  12. Detection panel for identification of twelve hemorrhagic viruses using real-time RT-PCR.

    Science.gov (United States)

    Fajfr, M; Neubauerová, V; Pajer, P; Kubíčková, P; Růžek, D

    2014-09-01

    Viral hemorrhagic fevers are caused by viruses from four viral families and develop diseases with high fatality rates. However, no commercial diagnostic assay for these pathogens is available. We developed real-time RT-PCR assays for viruses Ebola, Marburg, Lassa, Guanarito, Machupo, Junin, Sabiá, Seoul, Puumala, Hantaan, Crimean-Congo hemorrhagic fever virus and Rift Valley fever virus. The assays were optimized for identical reaction conditions and can be performed using several types of real-time PCR instruments, both capillary and plate, including a portable Ruggedized Advanced Pathogen Identification Device (R.A.P.I.D.) (Idaho Technology, Inc.). In combination with primers and probes from previously published studies, we present a simple system for rapid identification of hemorrhagic filoviruses, arenaviruses and bunyaviruses with sufficient sensitivity for first contact laboratory and diagnosis under field conditions.

  13. Memory Efficient VLSI Implementation of Real-Time Motion Detection System Using FPGA Platform

    Directory of Open Access Journals (Sweden)

    Sanjay Singh

    2017-06-01

    Full Text Available Motion detection is the heart of a potentially complex automated video surveillance system, intended to be used as a standalone system. Therefore, in addition to being accurate and robust, a successful motion detection technique must also be economical in the use of computational resources on selected FPGA development platform. This is because many other complex algorithms of an automated video surveillance system also run on the same platform. Keeping this key requirement as main focus, a memory efficient VLSI architecture for real-time motion detection and its implementation on FPGA platform is presented in this paper. This is accomplished by proposing a new memory efficient motion detection scheme and designing its VLSI architecture. The complete real-time motion detection system using the proposed memory efficient architecture along with proper input/output interfaces is implemented on Xilinx ML510 (Virtex-5 FX130T FPGA development platform and is capable of operating at 154.55 MHz clock frequency. Memory requirement of the proposed architecture is reduced by 41% compared to the standard clustering based motion detection architecture. The new memory efficient system robustly and automatically detects motion in real-world scenarios (both for the static backgrounds and the pseudo-stationary backgrounds in real-time for standard PAL (720 × 576 size color video.

  14. Real-time vibration-based structural damage detection using one-dimensional convolutional neural networks

    Science.gov (United States)

    Abdeljaber, Osama; Avci, Onur; Kiranyaz, Serkan; Gabbouj, Moncef; Inman, Daniel J.

    2017-02-01

    Structural health monitoring (SHM) and vibration-based structural damage detection have been a continuous interest for civil, mechanical and aerospace engineers over the decades. Early and meticulous damage detection has always been one of the principal objectives of SHM applications. The performance of a classical damage detection system predominantly depends on the choice of the features and the classifier. While the fixed and hand-crafted features may either be a sub-optimal choice for a particular structure or fail to achieve the same level of performance on another structure, they usually require a large computation power which may hinder their usage for real-time structural damage detection. This paper presents a novel, fast and accurate structural damage detection system using 1D Convolutional Neural Networks (CNNs) that has an inherent adaptive design to fuse both feature extraction and classification blocks into a single and compact learning body. The proposed method performs vibration-based damage detection and localization of the damage in real-time. The advantage of this approach is its ability to extract optimal damage-sensitive features automatically from the raw acceleration signals. Large-scale experiments conducted on a grandstand simulator revealed an outstanding performance and verified the computational efficiency of the proposed real-time damage detection method.

  15. Real Time Vision System for Obstacle Detection and Localization on FPGA

    OpenAIRE

    Alhamwi , Ali; Vandeportaele , Bertrand; Piat , Jonathan

    2015-01-01

    International audience; Obstacle detection is a mandatory function for a robot navigating in an indoor environment especially when interaction with humans is done in a cluttered environment. Commonly used vision-based solutions like SLAM (Simultaneous Localization and Mapping) or optical flow tend to be computation intensive and require powerful computation resources to meet low speed real-time constraints. Solutions using LIDAR (Light Detection And Ranging) sensors are more robust but not co...

  16. Comparison of Real Time Polymerase Chain Reaction with Microscopy and Antigen Detection Assay for the Diagnosis of Malaria

    International Nuclear Information System (INIS)

    Khan, S. A.; Ahmed, S.; Khan, F. A.; Shamshad, G. U.; Joyia, Z.; Mushahid, N.; Saeed, S.

    2013-01-01

    Objective: To determine the sensitivity of a real time polymerase chain reaction (PCR) for malaria diagnosis and to compare its accuracy with microscopy and an antigen based rapid diagnostic test (OptiMal). Study Design: Cross-sectional analytical study. Place and Duration of Study: Military Hospital, Armed Forces Institute of Transfusion and Armed Forces Institute of Pathology, Rawalpindi, from July to December 2011. Methodology: Venous blood samples of 300 clinically suspected patients of malaria were tested for malaria parasite by microscopy and OptiMal; and malaria parasite index was calculated for the positive samples. Plasmodium genus specific real time PCR was performed on all specimens, targeting small subunit rRNA gene. Diagnostic accuracy of three tests was compared and cost analysis was done. Results: Out of 300 patients, malaria parasite was detected in 110, 106 and 123 patients by microscopy, OptiMAL and PCR respectively. Real time PCR was 100% sensitive while microscopy and OptiMal had sensitivity of 89.4% and 86.2% respectively. All methods were 100% specific. The cost per test was calculated to be 0.2, 2.75 and 3.30 US dollar by microscopy, OptiMal and PCR respectively, excluding the once capital cost on PCR equipment. Conclusion: Genus specific real time PCR for the diagnosis of malaria was successfully established as a highly sensitive and affordable technology that should be incorporated in the diagnostic algorithm in this country. (author)

  17. Incremental Activation Detection for Real-Time fMRI Series Using Robust Kalman Filter

    Directory of Open Access Journals (Sweden)

    Liang Li

    2014-01-01

    Full Text Available Real-time functional magnetic resonance imaging (rt-fMRI is a technique that enables us to observe human brain activations in real time. However, some unexpected noises that emerged in fMRI data collecting, such as acute swallowing, head moving and human manipulations, will cause much confusion and unrobustness for the activation analysis. In this paper, a new activation detection method for rt-fMRI data is proposed based on robust Kalman filter. The idea is to add a variation to the extended kalman filter to handle the additional sparse measurement noise and a sparse noise term to the measurement update step. Hence, the robust Kalman filter is designed to improve the robustness for the outliers and can be computed separately for each voxel. The algorithm can compute activation maps on each scan within a repetition time, which meets the requirement for real-time analysis. Experimental results show that this new algorithm can bring out high performance in robustness and in real-time activation detection.

  18. Development of real-time PCR for detection and quantitation of Streptococcus parauberis.

    Science.gov (United States)

    Nguyen, T L; Lim, Y J; Kim, D-H; Austin, B

    2016-01-01

    Streptococcus parauberis is an increasing threat to aquaculture of olive flounder, Paralichthys olivaceus Temminck & Schlegel, in South Korea. We developed a real-time polymerase chain reaction (PCR) method using the TaqMan probe assay to detect and quantify S. parauberis by targeting the gyrB gene sequences, which are effective for molecular analysis of the genus Streptococcus. Our real-time PCR assay is capable of detecting 10 fg of genomic DNA per reaction. The intra- and interassay coefficient of variation (CV) values ranged from 0.42-1.95%, demonstrating that the assay has good reproducibility. There was not any cross-reactivity to Streptococcus iniae or to other streptococcal/lactococcal fish pathogens, such as S. agalactiae and Lactococcus garvieae, indicating that the assay is highly specific to S. parauberis. The results of the real-time PCR assay corresponded well to those of conventional culture assays for S. parauberis from inoculated tissue homogenates (r = 0.957; P < 0.05). Hence, this sensitive and specific real-time PCR is a valuable tool for diagnostic quantitation of S. parauberis in clinical samples. © 2014 John Wiley & Sons Ltd.

  19. Real-time PCR detection of Listeria monocytogenes in infant formula and lettuce following macrophage-based isolation and enrichment.

    Science.gov (United States)

    Day, J B; Basavanna, U

    2015-01-01

    To develop a rapid detection procedure for Listeria monocytogenes in infant formula and lettuce using a macrophage-based enrichment protocol and real-time PCR. A macrophage cell culture system was employed for the isolation and enrichment of L. monocytogenes from infant formula and lettuce for subsequent identification using real-time PCR. Macrophage monolayers were exposed to infant formula and lettuce contaminated with a serial dilution series of L. monocytogenes. As few as approx. 10 CFU ml(-1) or g(-1) of L. monocytogenes were detected in infant formula and lettuce after 16 h postinfection by real-time PCR. Internal positive PCR controls were utilized to eliminate the possibility of false-negative results. Co-inoculation with Listeria innocua did not reduce the L. monocytogenes detection sensitivity. Intracellular L. monocytogenes could also be isolated on Listeria selective media from infected macrophage lysates for subsequent confirmation. The detection method is highly sensitive and specific for L. monocytogenes in infant formula and lettuce and establishes a rapid identification time of 20 and 48 h for presumptive and confirmatory identification, respectively. The method is a promising alternative to many currently used q-PCR detection methods which employ traditional selective media for enrichment of contaminated food samples. Macrophage enrichment of L. monocytogenes eliminates PCR inhibitory food elements and contaminating food microflora which produce cleaner samples that increase the rapidity and sensitivity of detection. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

  20. Design of Wearable Breathing Sound Monitoring System for Real-Time Wheeze Detection

    Directory of Open Access Journals (Sweden)

    Shih-Hong Li

    2017-01-01

    Full Text Available In the clinic, the wheezing sound is usually considered as an indicator symptom to reflect the degree of airway obstruction. The auscultation approach is the most common way to diagnose wheezing sounds, but it subjectively depends on the experience of the physician. Several previous studies attempted to extract the features of breathing sounds to detect wheezing sounds automatically. However, there is still a lack of suitable monitoring systems for real-time wheeze detection in daily life. In this study, a wearable and wireless breathing sound monitoring system for real-time wheeze detection was proposed. Moreover, a breathing sounds analysis algorithm was designed to continuously extract and analyze the features of breathing sounds to provide the objectively quantitative information of breathing sounds to professional physicians. Here, normalized spectral integration (NSI was also designed and applied in wheeze detection. The proposed algorithm required only short-term data of breathing sounds and lower computational complexity to perform real-time wheeze detection, and is suitable to be implemented in a commercial portable device, which contains relatively low computing power and memory. From the experimental results, the proposed system could provide good performance on wheeze detection exactly and might be a useful assisting tool for analysis of breathing sounds in clinical diagnosis.

  1. A sensitive, reproducible, and economic real-time reverse transcription PCR detecting avian metapneumovirus subtypes A and B.

    Science.gov (United States)

    Franzo, G; Drigo, M; Lupini, C; Catelli, E; Laconi, A; Listorti, V; Bonci, M; Naylor, C J; Martini, M; Cecchinato, M

    2014-06-01

    Use of real-time PCR is increasing in the diagnosis of infectious disease due to its sensitivity, specificity, and speed of detection. These characteristics make it particularly suited for the diagnosis of viral infections, like avian metapneumovirus (AMPV), for which effective control benefits from continuously updated knowledge of the epidemiological situation. Other real-time reverse transcription (RT)-PCRs have been published based on highly specific fluorescent dye-labeled probes, but they have high initial cost, complex validation, and a marked susceptibility to the genetic variability of their target sequence. With this in mind, we developed and validated a SYBR Green I-based quantitative RT-PCR for the detection of the two most prevalent AMPV subtypes (i.e., subtypes A and B). The assay demonstrated an analytical sensitivity comparable with that of a previously published real-time RT-PCR and the ability to detect RNA equivalent to approximately 0.5 infectious doses for both A and B subtypes. The high efficiency and linearity between viral titer and crossing point displayed for both subtypes make it suited for viral quantification. Optimization of reaction conditions and the implementation of melting curve analysis guaranteed the high specificity of the assay. The stable melting temperature difference between the two subtypes indicated the possibility of subtyping through melting temperature analysis. These characteristics make our assay a sensitive, specific, and rapid tool, enabling contemporaneous detection, quantification, and discrimination of AMPV subtype A and B.

  2. Near real-time shadow detection and removal in aerial motion imagery application

    Science.gov (United States)

    Silva, Guilherme F.; Carneiro, Grace B.; Doth, Ricardo; Amaral, Leonardo A.; Azevedo, Dario F. G. de

    2018-06-01

    This work presents a method to automatically detect and remove shadows in urban aerial images and its application in an aerospace remote monitoring system requiring near real-time processing. Our detection method generates shadow masks and is accelerated by GPU programming. To obtain the shadow masks, we converted images from RGB to CIELCh model, calculated a modified Specthem ratio, and applied multilevel thresholding. Morphological operations were used to reduce shadow mask noise. The shadow masks are used in the process of removing shadows from the original images using the illumination ratio of the shadow/non-shadow regions. We obtained shadow detection accuracy of around 93% and shadow removal results comparable to the state-of-the-art while maintaining execution time under real-time constraints.

  3. SMA Diagnosis: Detection of SMN1 Deletion with Real-Time mCOP-PCR System Using Fresh Blood DNA.

    Science.gov (United States)

    Niba, Emma Tabe Eko; Ar Rochmah, Mawaddah; Harahap, Nur Imma Fatimah; Awano, Hiroyuki; Morioka, Ichiro; Iijima, Kazumoto; Saito, Toshio; Saito, Kayoko; Takeuchi, Atsuko; Lai, Poh San; Bouike, Yoshihiro; Nishio, Hisahide; Shinohara, Masakazu

    2017-12-18

    Spinal muscular atrophy (SMA) is one of the most common autosomal recessive disorders. The symptoms are caused by defects of lower motor neurons in the spinal cord. More than 95% of SMA patients are homozygous for survival motor neuron 1 (SMN1) deletion. We previously developed a screening system for SMN1 deletion based on a modified competitive oligonucleotide priming-PCR (mCOP-PCR) technique using dried blood spot (DBS) on filter paper. This system is convenient for mass screening in the large population and/or first-tier diagnostic method of the patients in the remote areas. However, this system was still time-consuming and effort-taking, because it required pre-amplification procedure to avoid non-specific amplification and gel-electrophoresis to detect the presence or absence of SMN1 deletion. When the fresh blood samples are used instead of DBS, or when the gel-electrophoresis is replaced by real-time PCR, we may have a simpler and more rapid diagnostic method for SMA. To establish a simpler and more rapid diagnostic method of SMN1 deletion using fresh blood DNA. DNA samples extracted from fresh blood and stored at 4 ℃ for 1 month. The samples were assayed using a real-time mCOP-PCR system without pre-amplification procedures. DNA samples had already been genotyped by PCR-restriction fragment length polymorphism (PCR-RFLP), showing the presence or absence of SMN1 exon 7. The DNA samples were directly subjected to the mCOP-PCR step. The amplification of mCOP-PCR was monitored in a real-time PCR apparatus. The genotyping results of the real-time mCOP-PCR system using fresh blood DNA were completely matched with those of PCR-RFLP. In this real-time mCOP-PCR system using fresh blood-DNA, it took only four hours from extraction of DNA to detection of the presence or absence of SMN1 deletion, while it took more than 12 hours in PCR-RFLP. Our real-time mCOP-PCR system using fresh blood DNA was rapid and accurate, suggesting it may be useful for the first

  4. Rapidly-steered single-element ultrasound for real-time volumetric imaging and guidance

    Science.gov (United States)

    Stauber, Mark; Western, Craig; Solek, Roman; Salisbury, Kenneth; Hristov, Dmitre; Schlosser, Jeffrey

    2016-03-01

    Volumetric ultrasound (US) imaging has the potential to provide real-time anatomical imaging with high soft-tissue contrast in a variety of diagnostic and therapeutic guidance applications. However, existing volumetric US machines utilize "wobbling" linear phased array or matrix phased array transducers which are costly to manufacture and necessitate bulky external processing units. To drastically reduce cost, improve portability, and reduce footprint, we propose a rapidly-steered single-element volumetric US imaging system. In this paper we explore the feasibility of this system with a proof-of-concept single-element volumetric US imaging device. The device uses a multi-directional raster-scan technique to generate a series of two-dimensional (2D) slices that were reconstructed into three-dimensional (3D) volumes. At 15 cm depth, 90° lateral field of view (FOV), and 20° elevation FOV, the device produced 20-slice volumes at a rate of 0.8 Hz. Imaging performance was evaluated using an US phantom. Spatial resolution was 2.0 mm, 4.7 mm, and 5.0 mm in the axial, lateral, and elevational directions at 7.5 cm. Relative motion of phantom targets were automatically tracked within US volumes with a mean error of -0.3+/-0.3 mm, -0.3+/-0.3 mm, and -0.1+/-0.5 mm in the axial, lateral, and elevational directions, respectively. The device exhibited a mean spatial distortion error of 0.3+/-0.9 mm, 0.4+/-0.7 mm, and -0.3+/-1.9 in the axial, lateral, and elevational directions. With a production cost near $1000, the performance characteristics of the proposed system make it an ideal candidate for diagnostic and image-guided therapy applications where form factor and low cost are paramount.

  5. Real-time patient survey data during routine clinical activities for rapid-cycle quality improvement.

    Science.gov (United States)

    Wofford, James Lucius; Campos, Claudia L; Jones, Robert E; Stevens, Sheila F

    2015-03-12

    Surveying patients is increasingly important for evaluating and improving health care delivery, but practical survey strategies during routine care activities have not been available. We examined the feasibility of conducting routine patient surveys in a primary care clinic using commercially available technology (Web-based survey creation, deployment on tablet computers, cloud-based management of survey data) to expedite and enhance several steps in data collection and management for rapid quality improvement cycles. We used a Web-based data management tool (survey creation, deployment on tablet computers, real-time data accumulation and display of survey results) to conduct four patient surveys during routine clinic sessions over a one-month period. Each survey consisted of three questions and focused on a specific patient care domain (dental care, waiting room experience, care access/continuity, Internet connectivity). Of the 727 available patients during clinic survey days, 316 patients (43.4%) attempted the survey, and 293 (40.3%) completed the survey. For the four 3-question surveys, the average time per survey was overall 40.4 seconds, with a range of 5.4 to 20.3 seconds for individual questions. Yes/No questions took less time than multiple choice questions (average 9.6 seconds versus 14.0). Average response time showed no clear pattern by order of questions or by proctor strategy, but monotonically increased with number of words in the question (30 words)-8.0, 11.8, 16.8, seconds, respectively. This technology-enabled data management system helped capture patient opinions, accelerate turnaround of survey data, with minimal impact on a busy primary care clinic. This new model of patient survey data management is feasible and sustainable in a busy office setting, supports and engages clinicians in the quality improvement process, and harmonizes with the vision of a learning health care system.

  6. Design and implement of infrared small target real-time detection system based on pipeline technology

    Science.gov (United States)

    Sun, Lihui; Wang, Yongzhong; He, Yongqiang

    2007-01-01

    The detection for motive small target in infrared image sequence has become a hot topic nowadays. Background suppress algorithm based on minim gradient median filter and temporal recursion target detection algorithm are introduced. On the basis of contents previously mentioned, a four stages pipeline structure infrared small target detection process system, which aims at characters of algorithm complexity, large amounts of data to process, high frame frequency and exigent real-time character in this kind of application, is designed and implemented. The logical structure of the system was introduced and the function and signals flows are programmed. The system is composed of two FPGA chips and two DSP chips of TI. According to the function of each part, the system is divided into image preprocess stage, target detection stage, track relation stage and image output stage. The experiment of running algorithms on the system presented in this paper proved that the system could meet acquisition and process of 50Hz 240x320 digital image and the system could real time detect small target with a signal-noise ratio more than 3 reliably. The system achieves the characters of large amount of memory, high real-time processing, excellent extension and favorable interactive interface.

  7. Rapid identification of ST131 Escherichia coli by a novel multiplex real-time allelic discrimination assay.

    Science.gov (United States)

    François, Patrice; Bonetti, Eve-Julie; Fankhauser, Carolina; Baud, Damien; Cherkaoui, Abdessalam; Schrenzel, Jacques; Harbarth, Stephan

    2017-09-01

    Escherichia coli sequence type 131 is increasingly described in severe hospital infections. We developed a rapid real-time allelic discrimination assay for the rapid identification of E. coli ST131 isolates. This rapid assay represents an affordable alternative to sequence-based strategies before completing characterization of potentially highly virulent isolates of E. coli. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Improved detection of canine Angiostrongylus vasorum infection using real-time PCR and indirect ELISA.

    Science.gov (United States)

    Jefferies, Ryan; Morgan, Eric R; Helm, Jenny; Robinson, Matthew; Shaw, Susan E

    2011-12-01

    This study reports the development of a real-time PCR assay and an indirect ELISA to improve on current detection of canine Angiostrongylus vasorum infection. A highly specific fluorescent probe-based, real-time PCR assay was developed to target the A. vasorum second internal transcribed spacer region and detected DNA in EDTA blood, lung tissue, broncho-alveolar larvage fluid, endotracheal mucus, pharyngeal swabs and faecal samples. PCR was fast (∼1 h), highly efficient when using EDTA blood samples, consistently detected a single molecule of parasite DNA and did not amplify DNA from other parasitic nematodes or definitive host species. An indirect ELISA was also developed using the soluble protein fraction from adult A. vasorum worms. Some cross-reactive antigen recognition was observed when tested against sera from dogs infected with Crenosoma vulpis (n = 8), Toxocara canis (n = 5) and Dirofilaria immitis (n = 5). This was largely overcome by setting the cut-off for a positive result at an appropriately high level. Field evaluation of the real-time PCR and ELISA was conducted by testing sera and EDTA blood from dogs with suspected A. vasorum infection (n = 148) and compared with the Baermann's larval migration test in faeces. Thirty-one dogs were positive by at least one test. Of these, 20 (65%) were detected by the Baermann method, 18 (58%) by blood PCR, 24 (77%) by ELISA and 28 (90%) by blood PCR and ELISA together. Combined testing using real-time PCR and ELISA therefore improved the detection rate of A. vasorum infection and holds promise for improved clinical diagnosis and epidemiological investigation.

  9. Development of quantitative real-time PCR for detection and enumeration of Enterobacteriaceae.

    Science.gov (United States)

    Takahashi, Hajime; Saito, Rumi; Miya, Satoko; Tanaka, Yuichiro; Miyamura, Natsumi; Kuda, Takashi; Kimura, Bon

    2017-04-04

    The family Enterobacteriaceae, members of which are widely distributed in the environment, includes many important human pathogens. In this study, a rapid real-time PCR method targeting rplP, coding for L16 protein, a component of the ribosome large subunit, was developed for enumerating Enterobacteriaceae strains, and its efficiency was evaluated using naturally contaminated food products. The rplP-targeted real-time PCR amplified Enterobacteriaceae species with Ct values of 14.0-22.8, whereas the Ct values for non-Enterobacteriaceae species were >30, indicating the specificity of this method for the Enterobacteriaceae. Using a calibration curve of Ct=-3.025 (log CFU/g)+37.35, which was calculated from individual plots of the cell numbers in different concentrations of 5 Enterobacteriaceae species, the rplP-targeted real-time PCR was applied to 51 food samples. A Enterobacteriaceae species in foods rapidly and accurately, and therefore, it can be used for the microbiological risk analysis of foods. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Rapid real-time PCR assay for culture and tissue identification of Geomyces destructans: the etiologic agent of bat geomycosis (white nose syndrome).

    Science.gov (United States)

    Chaturvedi, Sudha; Rudd, Robert J; Davis, April; Victor, Tanya R; Li, Xiaojiang; Appler, Kim A; Rajkumar, Sunanda S; Chaturvedi, Vishnu

    2011-10-01

    Geomyces destructans is the etiologic agent of bat geomycosis, commonly referred to as white nose syndrome (WNS). This infection has caused severe morbidity and mortality in little brown bats (Myotis lucifugus) and has also spread to other bat species with significant decline in the populations. Currently, G. destructans infection is identified by culture, ITS-PCR, and histopathology. We hypothesized that a real-time PCR assay would considerably improve detection of G. destructans in bats. The 100 bp sequence of the Alpha-L-Rhamnosidase gene was validated as a target for real-time PCR. The assay sensitivity was determined from serial dilution of DNA extracted from G. destructans conidia (5 × 10(-1)-5 × 10(7)), and the specificity was tested using DNA from 30 closely and distantly related fungi and 5 common bacterial pathogens. The real-time PCR assay was highly sensitive with detection limit of two G. destructans conidia per reaction at 40 PCR cycles. The assay was also highly specific as none of the other fungal or bacterial DNA cross-reacted in the real-time PCR assay. One hundred and forty-seven bat tissue samples, suspected of infection with G. destructans, were used to compare the real-time PCR assay to other methods employed for the detection of G. destructans. Real-time PCR was highly sensitive with 80 of 147 (55%) samples testing positive for G. destructans DNA. In comparison, histopathology examination revealed 64/147 (44%) positive samples. The internal transcribed spacer (ITS)-PCR yielded positive amplicon for G. destructans from 37 tissue samples (25%). The least sensitive assay was the fungal culture with only 17 tissue samples (12%) yielding G. destructans in culture. The data suggested that the real-time PCR assay is highly promising for rapid, sensitive, and specific identification of G. destructans. Further trials and inter-laboratory comparisons of this novel assay are recommended to improve the diagnosis of bat geomycosis.

  11. Direct Analysis in Real-time Mass Spectrometry for Rapid Identification of Traditional Chinese Medicines with Coumarins as Primary Characteristics.

    Science.gov (United States)

    Chen, Zhiyong; Yang, Yuanyuan; Tao, Hongxun; Liao, Liping; Li, Ye; Zhang, Zijia

    2017-05-01

    The increasing popularity of traditional Chinese medicines (TCMs) necessitates rapid and reliable methods for controlling their quality. Direct analysis in real-time mass spectrometry (DART-MS) represents a novel approach to analysing TCMs. To develop a quick and reliable method of identifying TCMs with coumarins as primary characteristics. DART-MS coupled with ion trap mass spectrometry was employed to rapidly identify TCMs with coumarins as primary characteristics and to explore the ionisation mechanisms of simple coumarins, furocoumarins and pyranocoumarins in detail. With minimal sample pretreatment, mass spectra of Fraxini Cortex, Angelicae Pubescentis Radix, Peucedani Radix and Psoraleae Fructus samples were obtained within seconds. The operating parameters of the DART ion source (e.g. grid electrode voltage and ionisation gas temperature) were carefully investigated to obtain high-quality mass spectra. The mass spectra of samples and DART-MS/MS spectra of marker compounds were used to identify sample materials. Successful authentication was achieved by analysing the same materials of different origins. Some simple coumarins, furocoumarins and pyranocoumarins can be directly detected by DART-MS as marker compounds. Our results demonstrated that DART-MS can provide a rapid and reliable method for the identification of TCMs containing different configurations of coumarins; the method may also be applicable to other plants. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  12. A Novel Reverse-Transcriptase Real-Time PCR Method for Quantification of Viable Vibrio Parahemolyticus in Raw Shrimp Based on a Rapid Construction of Standard Curve Method

    OpenAIRE

    Mengtong Jin; Haiquan Liu; Wenshuo Sun; Qin Li; Zhaohuan Zhang; Jibing Li; Yingjie Pan; Yong Zhao

    2015-01-01

    Vibrio parahemolyticus is an important pathogen that leads to food illness associated seafood. Therefore, rapid and reliable methods to detect and quantify the total viable V. parahaemolyticus in seafood are needed. In this assay, a RNA-based real-time reverse-transcriptase PCR (RT-qPCR) without an enrichment step has been developed for detection and quantification of the total viable V. parahaemolyticus in shrimp. RNA standards with the target segments were synthesized in vitro with T7 RNA p...

  13. Integration of Simulink, MARTe and MDSplus for rapid development of real-time applications

    Energy Technology Data Exchange (ETDEWEB)

    Manduchi, G., E-mail: gabriele.manduchi@igi.cnr.it [Consorzio RFX (CNR, ENEA, INFN, Università di Padova, Acciaierie Venete SpA), Padova (Italy); Luchetta, A.; Taliercio, C. [Consorzio RFX (CNR, ENEA, INFN, Università di Padova, Acciaierie Venete SpA), Padova (Italy); Neto, A.; Sartori, F. [Fusion for Energy, Barcelona (Spain); De Tommasi, G. [Fusion for Energy, Barcelona (Spain); Consorzio CREATE/DIETI, Università degli Studi di Napoli Federico II, Via Claudio 21, 80125 Napoli (Italy)

    2015-10-15

    Highlights: • The integration of two frameworks for real-time control and data acquisition is described. • The integration may significantly fasten the development of system components. • The system includes also a code generator for the integration of code written in Simulink. • A real-time control systemcan be implemented without the need of writing any line of code. - Abstract: Simulink is a graphical data flow programming tool for modeling and simulating dynamic systems. A component of Simulink, called Simulink Coder, generates C code from Simulink diagrams. MARTe is a framework for the implementation of real-time systems, currently in use in several fusion experiments. MDSplus is a framework widely used in the fusion community for the management of data. The three systems provide a solution to different facets of the same process, that is, real-time plasma control development. Simulink diagrams will describe the algorithms used in control, which will be implemented as MARTe GAMs and which will use parameters read from and produce results written to MDSplus pulse files. The three systems have been integrated in order to provide a tool suitable to speed up the development of real-time control applications. In particular, it will be shown how from a Simulink diagram describing a given algorithm to be used in a control system, it is possible to generate in an automated way the corresponding MARTe and MDSplus components that can be assembled to implement the target system.

  14. Integration of Simulink, MARTe and MDSplus for rapid development of real-time applications

    International Nuclear Information System (INIS)

    Manduchi, G.; Luchetta, A.; Taliercio, C.; Neto, A.; Sartori, F.; De Tommasi, G.

    2015-01-01

    Highlights: • The integration of two frameworks for real-time control and data acquisition is described. • The integration may significantly fasten the development of system components. • The system includes also a code generator for the integration of code written in Simulink. • A real-time control systemcan be implemented without the need of writing any line of code. - Abstract: Simulink is a graphical data flow programming tool for modeling and simulating dynamic systems. A component of Simulink, called Simulink Coder, generates C code from Simulink diagrams. MARTe is a framework for the implementation of real-time systems, currently in use in several fusion experiments. MDSplus is a framework widely used in the fusion community for the management of data. The three systems provide a solution to different facets of the same process, that is, real-time plasma control development. Simulink diagrams will describe the algorithms used in control, which will be implemented as MARTe GAMs and which will use parameters read from and produce results written to MDSplus pulse files. The three systems have been integrated in order to provide a tool suitable to speed up the development of real-time control applications. In particular, it will be shown how from a Simulink diagram describing a given algorithm to be used in a control system, it is possible to generate in an automated way the corresponding MARTe and MDSplus components that can be assembled to implement the target system.

  15. Real-time Alarm Monitoring System for Detecting Driver Fatigue in Wireless Areas

    Directory of Open Access Journals (Sweden)

    Rongrong Fu

    2017-04-01

    Full Text Available The purpose of this paper was to develop a real-time alarm monitoring system that can detect the fatigue driving state through wireless communication. The drivers’ electroencephalogram (EEG signals were recorded from occipital electrodes. Seven EEG rhythms with different frequency bands as gamma, hbeta, beta, sigma, alpha, theta and delta waves were extracted. They were simultaneously assessed using relative operating characteristic (ROC curves and grey relational analysis to select one as the fatigue feature. The research results showed that the performance of theta wave was the best one. Therefore, theta wave was used as fatigue feature in the following alarm device. The real-time alarm monitoring system based on the result has been developed, once the threshold was settled by using the data of the first ten minutes driving period. The developed system can detect driver fatigue and give alarm to indicate the onset of fatigue automatically.

  16. Real-time billboard trademark detection and recognition in sports video

    Science.gov (United States)

    Bu, Jiang; Lao, Song-Yan; Bai, Liang

    2013-03-01

    Nowadays, different applications like automatic video indexing, keyword based video search and TV commercials can be developed by detecting and recognizing the billboard trademark. We propose a hierarchical solution for real-time billboard trademark recognition in various sports video, billboard frames are detected in the first level, fuzzy decision tree with easily-computing features are employed to accelerate the process, while in the second level, color and regional SIFT features are combined for the first time to describe the appearance of trademarks, and the shared nearest neighbor (SNN) clustering with x2 distance is utilized instead of traditional K-means clustering to construct the SIFT vocabulary, at last, Latent Semantic Analysis (LSA) based SIFT vocabulary matching is performed on the template trademark and the candidate regions in billboard frame. The preliminary experiments demonstrate the effectiveness of the hierarchical solution, and real time constraints are also met by our solution.

  17. Real-time PCR detection of aldoxime dehydratase genes in nitrile-degrading microorganisms.

    Science.gov (United States)

    Dooley-Cullinane, Tríona Marie; O'Reilly, Catherine; Coffey, Lee

    2017-02-01

    Aldoxime dehydratase catalyses the conversion of aldoximes to their corresponding nitriles. Utilization of the aldoxime-nitrile metabolising enzyme pathway can facilitate the move towards a greener chemistry. In this work, a real-time PCR assay was developed for the detection of aldoxime dehydratase genes in aldoxime/nitrile metabolising microorganisms which have been purified from environmental sources. A conventional PCR assay was also designed allowing gene confirmation via sequencing. Aldoxime dehydratase genes were identified in 30 microorganisms across 11 genera including some not previously shown to harbour the gene. The assay displayed a limit of detection of 1 pg/μL DNA or 7 CFU/reaction. This real-time PCR assay should prove valuable in the high-throughput screening of micro-organisms for novel aldoxime dehydratase genes towards pharmaceutical and industrial applications.

  18. Real-time DSP implementation for MRF-based video motion detection.

    Science.gov (United States)

    Dumontier, C; Luthon, F; Charras, J P

    1999-01-01

    This paper describes the real time implementation of a simple and robust motion detection algorithm based on Markov random field (MRF) modeling, MRF-based algorithms often require a significant amount of computations. The intrinsic parallel property of MRF modeling has led most of implementations toward parallel machines and neural networks, but none of these approaches offers an efficient solution for real-world (i.e., industrial) applications. Here, an alternative implementation for the problem at hand is presented yielding a complete, efficient and autonomous real-time system for motion detection. This system is based on a hybrid architecture, associating pipeline modules with one asynchronous module to perform the whole process, from video acquisition to moving object masks visualization. A board prototype is presented and a processing rate of 15 images/s is achieved, showing the validity of the approach.

  19. Versatile real-time interferometer phase-detection system using high-speed digital techniques

    International Nuclear Information System (INIS)

    Mendell, D.S.; Willett, G.W.

    1977-01-01

    This paper describes the basic design and philosophy of a versatile real-time interferometer phase-detection system to be used on the 2XIIB and TMX magnetic-fusion experiments at Lawrence Livermore Laboratory. This diagnostics system is a satellite to a host computer and uses high-speed emitter-coupled logic techniques to derive data on real-time phase relationships. The system's input signals can be derived from interferometer outputs over a wide range of reference frequencies. An LSI-11 microcomputer is the interface between the high-speed phase-detection logic, buffer memory, human interaction, and host computer. Phase data on a storage CRT is immediately displayed after each experimental fusion shot. An operator can interrogate this phase data more closely from an interactive control panel, and the host computer can be simultaneously examining the system's buffer memory or arming the system for the next shot

  20. Rapid qualitative urinary tract infection pathogen identification by SeptiFast real-time PCR.

    Directory of Open Access Journals (Sweden)

    Lutz E Lehmann

    2011-02-01

    Full Text Available Urinary tract infections (UTI are frequent in outpatients. Fast pathogen identification is mandatory for shortening the time of discomfort and preventing serious complications. Urine culture needs up to 48 hours until pathogen identification. Consequently, the initial antibiotic regimen is empirical.To evaluate the feasibility of qualitative urine pathogen identification by a commercially available real-time PCR blood pathogen test (SeptiFast® and to compare the results with dipslide and microbiological culture.Pilot study with prospectively collected urine samples.University hospital.82 prospectively collected urine samples from 81 patients with suspected UTI were included. Dipslide urine culture was followed by microbiological pathogen identification in dipslide positive samples. In parallel, qualitative DNA based pathogen identification (SeptiFast® was performed in all samples.61 samples were SeptiFast® positive, whereas 67 samples were dipslide culture positive. The inter-methodological concordance of positive and negative findings in the gram+, gram- and fungi sector was 371/410 (90%, 477/492 (97% and 238/246 (97%, respectively. Sensitivity and specificity of the SeptiFast® test for the detection of an infection was 0.82 and 0.60, respectively. SeptiFast® pathogen identifications were available at least 43 hours prior to culture results.The SeptiFast® platform identified bacterial DNA in urine specimens considerably faster compared to conventional culture. For UTI diagnosis sensitivity and specificity is limited by its present qualitative setup which does not allow pathogen quantification. Future quantitative assays may hold promise for PCR based UTI pathogen identification as a supplementation of conventional culture methods.

  1. Comprehensive and Rapid Real-Time PCR Analysis of 21 Foodborne Outbreaks

    Directory of Open Access Journals (Sweden)

    Hiroshi Fukushima

    2009-01-01

    Full Text Available A set of four duplex SYBR Green I PCR (SG-PCR assay combined with DNA extraction using QIAamp DNA Stool Mini kit was evaluated for the detection of foodborne bacteria from 21 foodborne outbreaks. The causative pathogens were detected in almost all cases in 2 hours or less. The first run was for the detection of 8 main foodborne pathogens in 5 stool specimens within 2 hours and the second run was for the detection of other unusual suspect pathogens within a further 45 minutes. After 2 to 4 days, the causative agents were isolated and identified. The results proved that for comprehensive and rapid molecular diagnosis in foodborne outbreaks, Duplex SG-PCR assay is not only very useful, but is also economically viable for one-step differentiation of causative pathogens in fecal specimens obtained from symptomatic patients. This then allows for effective diagnosis and management of foodborne outbreaks.

  2. Real-time detection and discrimination of visual perception using electrocorticographic signals

    Science.gov (United States)

    Kapeller, C.; Ogawa, H.; Schalk, G.; Kunii, N.; Coon, W. G.; Scharinger, J.; Guger, C.; Kamada, K.

    2018-06-01

    Objective. Several neuroimaging studies have demonstrated that the ventral temporal cortex contains specialized regions that process visual stimuli. This study investigated the spatial and temporal dynamics of electrocorticographic (ECoG) responses to different types and colors of visual stimulation that were presented to four human participants, and demonstrated a real-time decoder that detects and discriminates responses to untrained natural images. Approach. ECoG signals from the participants were recorded while they were shown colored and greyscale versions of seven types of visual stimuli (images of faces, objects, bodies, line drawings, digits, and kanji and hiragana characters), resulting in 14 classes for discrimination (experiment I). Additionally, a real-time system asynchronously classified ECoG responses to faces, kanji and black screens presented via a monitor (experiment II), or to natural scenes (i.e. the face of an experimenter, natural images of faces and kanji, and a mirror) (experiment III). Outcome measures in all experiments included the discrimination performance across types based on broadband γ activity. Main results. Experiment I demonstrated an offline classification accuracy of 72.9% when discriminating among the seven types (without color separation). Further discrimination of grey versus colored images reached an accuracy of 67.1%. Discriminating all colors and types (14 classes) yielded an accuracy of 52.1%. In experiment II and III, the real-time decoder correctly detected 73.7% responses to face, kanji and black computer stimuli and 74.8% responses to presented natural scenes. Significance. Seven different types and their color information (either grey or color) could be detected and discriminated using broadband γ activity. Discrimination performance maximized for combined spatial-temporal information. The discrimination of stimulus color information provided the first ECoG-based evidence for color-related population

  3. Real-time instrument-failure detection in the LOFT pressurizer using functional redundancy

    International Nuclear Information System (INIS)

    Tylee, J.L.

    1982-07-01

    The functional redundancy approach to detecting instrument failures in a pressurized water reactor (PWR) pressurizer is described and evaluated. This real-time method uses a bank of Kalman filters (one for each instrument) to generate optimal estimates of the pressurizer state. By performing consistency checks between the output of each filter, failed instruments can be identified. Simulation results and actual pressurizer data are used to demonstrate the capabilities of the technique

  4. A compact multifunctional microfluidic platform for exploring cellular dynamics in real-time using electrochemical detection

    DEFF Research Database (Denmark)

    Zor, Kinga; Heiskanen, Arto; Caviglia, Claudia

    2014-01-01

    and electrochemical analysis platform with in-built fluid handling and detection, enabling complete cell based assays comprising on-line electrode cleaning, sterilization, surface functionalization, cell seeding, cultivation and electrochemical real-time monitoring of cellular dynamics. To demonstrate the versatility...... capability. The here presented platform is aimed at applications utilizing cell based assays, ranging from e.g. monitoring of drug effects in pharmacological studies, characterization of neural stem cell differentiation, and screening of genetically modified microorganisms to environmental monitoring....

  5. Deep Learning for real-time gravitational wave detection and parameter estimation: Results with Advanced LIGO data

    Science.gov (United States)

    George, Daniel; Huerta, E. A.

    2018-03-01

    The recent Nobel-prize-winning detections of gravitational waves from merging black holes and the subsequent detection of the collision of two neutron stars in coincidence with electromagnetic observations have inaugurated a new era of multimessenger astrophysics. To enhance the scope of this emergent field of science, we pioneered the use of deep learning with convolutional neural networks, that take time-series inputs, for rapid detection and characterization of gravitational wave signals. This approach, Deep Filtering, was initially demonstrated using simulated LIGO noise. In this article, we present the extension of Deep Filtering using real data from LIGO, for both detection and parameter estimation of gravitational waves from binary black hole mergers using continuous data streams from multiple LIGO detectors. We demonstrate for the first time that machine learning can detect and estimate the true parameters of real events observed by LIGO. Our results show that Deep Filtering achieves similar sensitivities and lower errors compared to matched-filtering while being far more computationally efficient and more resilient to glitches, allowing real-time processing of weak time-series signals in non-stationary non-Gaussian noise with minimal resources, and also enables the detection of new classes of gravitational wave sources that may go unnoticed with existing detection algorithms. This unified framework for data analysis is ideally suited to enable coincident detection campaigns of gravitational waves and their multimessenger counterparts in real-time.

  6. Development of a real-time PCR to detect Demodex canis DNA in different tissue samples.

    Science.gov (United States)

    Ravera, Ivan; Altet, Laura; Francino, Olga; Bardagí, Mar; Sánchez, Armand; Ferrer, Lluís

    2011-02-01

    The present study reports the development of a real-time polymerase chain reaction (PCR) to detect Demodex canis DNA on different tissue samples. The technique amplifies a 166 bp of D. canis chitin synthase gene (AB 080667) and it has been successfully tested on hairs extracted with their roots and on formalin-fixed paraffin embedded skin biopsies. The real-time PCR amplified on the hairs of all 14 dogs with a firm diagnosis of demodicosis and consistently failed to amplify on negative controls. Eleven of 12 skin biopsies with a morphologic diagnosis of canine demodicosis were also positive. Sampling hairs on two skin points (lateral face and interdigital skin), D. canis DNA was detected on nine of 51 healthy dogs (17.6%) a much higher percentage than previously reported with microscopic studies. Furthermore, it is foreseen that if the number of samples were increased, the percentage of positive dogs would probably also grow. Moreover, in four of the six dogs with demodicosis, the samples taken from non-lesioned skin were positive. This finding, if confirmed in further studies, suggests that demodicosis is a generalized phenomenon in canine skin, due to proliferation of local mite populations, even though macroscopic lesions only appear in certain areas. The real-time PCR technique to detect D. canis DNA described in this work is a useful tool to advance our understanding of canine demodicosis.

  7. Classification of acoustic emission signals for drive systems coupling crack detection in semi-real time

    International Nuclear Information System (INIS)

    Godinez, V.; Shu, F.; Finlayson, R.; O'Donnell, B.; Anastasopoulos, A.; Tsimogiannis, A.

    2004-01-01

    Early detection of mechanical failure in helicopter drive train components is a key safety and economical issue with both military and civil sectors of aviation. Of these components, couplings are particularly critical. The objective of this work is to demonstrate the feasibility of designing and developing a reliable, real time monitoring methodology based on Supervised Pattern Recognition (SPR) for early detection of cracks in couplings used in helicopter and engine drive systems. Within this framework, a portable Acoustic Emission (AE) system was used, equipped with a semi-real time SPR software package. Results from AE tests performed in a gearbox-testing bench at different speeds and different torque values are presented. These results indicate that the energy content of different frequency bands in the AE signals power spectra is strongly correlated with the introduction of EDM notches in the main gear. Further tests indicate that a strong shift in the frequency of the AE signals is observed after spalling occurred in the pinion gear. The variation of displacement and velocity between signal classes are discussed as a potential feature in characterizing crack severity. Finally, a scope of the work for optimizing the methodology in detecting and evaluating coupling cracking in real time will be presented. (author)

  8. Real-time detection and elimination of nonorthogonality error in interference fringe processing

    International Nuclear Information System (INIS)

    Hu Haijiang; Zhang Fengdeng

    2011-01-01

    In the measurement system of interference fringe, the nonorthogonality error is a main error source that influences the precision and accuracy of the measurement system. The detection and elimination of the error has been an important target. A novel method that only uses the cross-zero detection and the counting is proposed to detect and eliminate the nonorthogonality error in real time. This method can be simply realized by means of the digital logic device, because it does not invoke trigonometric functions and inverse trigonometric functions. And it can be widely used in the bidirectional subdivision systems of a Moire fringe and other optical instruments.

  9. Quantitative Real-time PCR detection of putrescine-producing Gram-negative bacteria

    Directory of Open Access Journals (Sweden)

    Kristýna Maršálková

    2017-01-01

    Full Text Available Biogenic amines are indispensable components of living cells; nevertheless these compounds could be toxic for human health in higher concentrations. Putrescine is supposed to be the major biogenic amine associated with microbial food spoilage. Development of reliable, fast and culture-independent molecular methods to detect bacteria producing biogenic amines deserves the attention, especially of the food industry in purpose to protect health. The objective of this study was to verify the newly designed primer sets for detection of two inducible genes adiA and speF together in Salmonella enterica and Escherichia coli genome by Real-time PCR. These forenamed genes encode enzymes in the metabolic pathway which leads to production of putrescine in Gram-negative bacteria. Moreover, relative expression of these genes was studied in E. coli CCM 3954 strain using Real-time PCR. In this study, sets of new primers for the detection two inducible genes (speF and adiA in Salmonella enterica and E. coli by Real-time PCR were designed and tested. Amplification efficiency of a Real-time PCR was calculated from the slope of the standard curves (adiA, speF, gapA. An efficiency in a range from 95 to 105 % for all tested reactions was achieved. The gene expression (R of adiA and speF genes in E. coli was varied depending on culture conditions. The highest gene expression of adiA and speF was observed at 6, 24 and 36 h (RadiA ~ 3, 5, 9; RspeF ~11, 10, 9; respectively after initiation of growth of this bacteria in nutrient broth medium enchired with amino acids. The results show that these primers could be used for relative quantification analysis of E. coli.

  10. On-site detection of Xylella fastidiosa in host plants and in “spy insects” using the real-time loop-mediated isothermal amplification method

    Directory of Open Access Journals (Sweden)

    Thaer YASEEN

    2015-12-01

    Full Text Available A recent severe outbreak of Xylella fastidiosa associated with ‘olive quick decline syndrome’ (OQDS was reported in Apulia (Southern Italy. In this study an on-site real-time loop-mediated isothermal amplification (real-time LAMP was developed for detecting X. fastidiosa in host plants and insects. A marked simplification of the DNA extraction procedure was obtained by heating the samples in a portable Smart-Dart device and using an optimized enhancer reaction buffer. The connection to a tablet or Smartphone allowed to visualize the results of the reaction in real time. Compared to PCR and ELISA, with which it showed comparable results in terms of sensitivity and reliability in the X. fastidiosa detection, this simplified real-time LAMP procedure proved to be “user friendly”, displaying the advantages to be an on-site detection method of easy handling, rapid execution and low cost.

  11. Development of a Real-Time Resistance Measurement for Vibrio parahaemolyticus Detection by the Lecithin-Dependent Hemolysin Gene

    Science.gov (United States)

    Xiang, Guiming; Pu, Xiaoyun; Jiang, Dongneng; Liu, Linlin; Liu, Chang; Liu, Xiaobo

    2013-01-01

    The marine bacterium Vibrio parahaemolyticus (V. parahaemolyticus) causes gastroenteritis in humans via the ingestion of raw or undercooked contaminated seafood, and early diagnosis and prompt treatment are important for the prevention of V. parahaemolyticus-related diseases. In this study, a real-time resistance measurement based on loop-mediated isothermal amplification (LAMP), electrochemical ion bonding (Crystal violet and Mg2+), real-time monitoring, and derivative analysis was developed. V. parahaemolyticus DNA was first amplified by LAMP, and the products (DNA and pyrophosphate) represented two types of negative ions that could combine with a positive dye (Crystal violet) and positive ions (Mg2+) to increase the resistance of the reaction liquid. This resistance was measured in real-time using a specially designed resistance electrode, thus permitting the quantitative detection of V. parahaemolyticus. The results were obtained in 1–2 hours, with a minimum bacterial density of 10 CFU.mL−1 and high levels of accuracy (97%), sensitivity (96.08%), and specificity (97.96%) when compared to cultivation methods. Therefore, this simple and rapid method has a potential application in the detection of V. parahaemolyticus on a gene chip or in point-of-care testing. PMID:23991096

  12. High throughput detection of Coxiella burnetii by real-time PCR with internal control system and automated DNA preparation

    Directory of Open Access Journals (Sweden)

    Kramme Stefanie

    2008-05-01

    Full Text Available Abstract Background Coxiella burnetii is the causative agent of Q-fever, a widespread zoonosis. Due to its high environmental stability and infectivity it is regarded as a category B biological weapon agent. In domestic animals infection remains either asymptomatic or presents as infertility or abortion. Clinical presentation in humans can range from mild flu-like illness to acute pneumonia and hepatitis. Endocarditis represents the most common form of chronic Q-fever. In humans serology is the gold standard for diagnosis but is inadequate for early case detection. In order to serve as a diagnostic tool in an eventual biological weapon attack or in local epidemics we developed a real-time 5'nuclease based PCR assay with an internal control system. To facilitate high-throughput an automated extraction procedure was evaluated. Results To determine the minimum number of copies that are detectable at 95% chance probit analysis was used. Limit of detection in blood was 2,881 copies/ml [95%CI, 2,188–4,745 copies/ml] with a manual extraction procedure and 4,235 copies/ml [95%CI, 3,143–7,428 copies/ml] with a fully automated extraction procedure, respectively. To demonstrate clinical application a total of 72 specimens of animal origin were compared with respect to manual and automated extraction. A strong correlation between both methods was observed rendering both methods suitable. Testing of 247 follow up specimens of animal origin from a local Q-fever epidemic rendered real-time PCR more sensitive than conventional PCR. Conclusion A sensitive and thoroughly evaluated real-time PCR was established. Its high-throughput mode may show a useful approach to rapidly screen samples in local outbreaks for other organisms relevant for humans or animals. Compared to a conventional PCR assay sensitivity of real-time PCR was higher after testing samples from a local Q-fever outbreak.

  13. Standardisation and evaluation of a quantitative multiplex real-time PCR assay for the rapid identification of Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    Feroze Ahmed Ganaie

    2015-01-01

    Full Text Available Rapid diagnosis of Streptococcus pneumoniae can play a significant role in decreasing morbidity and mortality of infection. The accurate diagnosis of pneumococcal disease is hampered by the difficulties in growing the isolates from clinical specimens and also by misidentification. Molecular methods have gained popularity as they offer improvement in the detection of causative pathogens with speed and ease. The present study aims at validating and standardising the use of 4 oligonucleotide primer-probe sets (pneumolysin [ply], autolysin [lytA], pneumococcal surface adhesion A [psaA] and Spn9802 [DNA fragment] in a single-reaction mixture for the detection and discrimination of S. pneumoniae. Here, we validate a quantitative multiplex real-time PCR (qmPCR assay with a panel consisting of 43 S. pneumoniae and 29 non-pneumococcal isolates, 20 culture positive, 26 culture negative and 30 spiked serum samples. A standard curve was obtained using S. pneumoniae ATCC 49619 strain and glyceraldehyde 3-phosphate dehydrogenase (GAPDH gene was used as an endogenous internal control. The experiment showed high sensitivity with lower limit of detection equivalent to 4 genome copies/µl. The efficiency of the reaction was 100% for ply, lytA, Spn9802 and 97% for psaA. The test showed sensitivity and specificity of 100% with culture isolates and serum specimens. This study demonstrates that qmPCR analysis of sera using 4 oligonucleotide primers appears to be an appropriate method for the genotypic identification of S. pneumoniae infection.

  14. Implementation of a FPGA-Based Feature Detection and Networking System for Real-time Traffic Monitoring

    OpenAIRE

    Chen, Jieshi; Schafer, Benjamin Carrion; Ho, Ivan Wang-Hei

    2016-01-01

    With the growing demand of real-time traffic monitoring nowadays, software-based image processing can hardly meet the real-time data processing requirement due to the serial data processing nature. In this paper, the implementation of a hardware-based feature detection and networking system prototype for real-time traffic monitoring as well as data transmission is presented. The hardware architecture of the proposed system is mainly composed of three parts: data collection, feature detection,...

  15. Evaluation of a multiplex real-time PCR assay for the detection of respiratory viruses in clinical specimens.

    Science.gov (United States)

    Rheem, Insoo; Park, Joowon; Kim, Tae-Hyun; Kim, Jong Wan

    2012-11-01

    In this study, we evaluated the analytical performance and clinical potential of a one-step multiplex real-time PCR assay for the simultaneous detection of 14 types of respiratory viruses using the AdvanSure RV real-time PCR Kit (LG Life Sciences, Korea). Three hundred and twenty clinical specimens were tested with the AdvanSure RV real-time PCR Kit and conventional multiplex reverse transcription (RT)-PCR assay. The assay results were analyzed and the one-step AdvanSure RV real-time PCR Kit was compared with the conventional multiplex RT-PCR assay with respect to the sensitivity and specificity of the detection of respiratory viruses. The limit of detection (LOD) was 1.31 plaque-forming units (PFU)/mL for human rhinoviruses (hRVs), 4.93 PFU/mL for human coronavirus HCoV-229E/NL63, 2.67 PFU/mL for human coronavirus HCoV-OC43, 18.20 PFU/mL for parainfluenza virus 1 (PIV)-1, 24.57 PFU/mL for PIV-2, 1.73 PFU/mL for PIV-3, 1.79 PFU/mL for influenza virus group (Flu) A, 59.51 PFU/mL for FluB, 5.46 PFU/mL for human respiratory syncytial virus (hRSV)-A, 17.23 PFU/mL for hRSV-B, 9.99 PFU/mL for human adenovirus (ADVs). The cross-reactivity test for this assay against 23 types of non-respiratory viruses showed negative results for all viruses tested. The agreement between the one-step AdvanSure multiplex real-time PCR assay and the conventional multiplex RT-PCR assay was 98%. The one-step AdvanSure RV multiplex real-time PCR assay is a simple assay with high potential for specific, rapid and sensitive laboratory diagnosis of respiratory viruses compared to conventional multiplex RT-PCR.

  16. A wearable smartphone-based platform for real-time cardiovascular disease detection via electrocardiogram processing.

    Science.gov (United States)

    Oresko, Joseph J; Duschl, Heather; Cheng, Allen C

    2010-05-01

    Cardiovascular disease (CVD) is the single leading cause of global mortality and is projected to remain so. Cardiac arrhythmia is a very common type of CVD and may indicate an increased risk of stroke or sudden cardiac death. The ECG is the most widely adopted clinical tool to diagnose and assess the risk of arrhythmia. ECGs measure and display the electrical activity of the heart from the body surface. During patients' hospital visits, however, arrhythmias may not be detected on standard resting ECG machines, since the condition may not be present at that moment in time. While Holter-based portable monitoring solutions offer 24-48 h ECG recording, they lack the capability of providing any real-time feedback for the thousands of heart beats they record, which must be tediously analyzed offline. In this paper, we seek to unite the portability of Holter monitors and the real-time processing capability of state-of-the-art resting ECG machines to provide an assistive diagnosis solution using smartphones. Specifically, we developed two smartphone-based wearable CVD-detection platforms capable of performing real-time ECG acquisition and display, feature extraction, and beat classification. Furthermore, the same statistical summaries available on resting ECG machines are provided.

  17. Deep Learning for Real-Time Capable Object Detection and Localization on Mobile Platforms

    Science.gov (United States)

    Particke, F.; Kolbenschlag, R.; Hiller, M.; Patiño-Studencki, L.; Thielecke, J.

    2017-10-01

    Industry 4.0 is one of the most formative terms in current times. Subject of research are particularly smart and autonomous mobile platforms, which enormously lighten the workload and optimize production processes. In order to interact with humans, the platforms need an in-depth knowledge of the environment. Hence, it is required to detect a variety of static and non-static objects. Goal of this paper is to propose an accurate and real-time capable object detection and localization approach for the use on mobile platforms. A method is introduced to use the powerful detection capabilities of a neural network for the localization of objects. Therefore, detection information of a neural network is combined with depth information from a RGB-D camera, which is mounted on a mobile platform. As detection network, YOLO Version 2 (YOLOv2) is used on a mobile robot. In order to find the detected object in the depth image, the bounding boxes, predicted by YOLOv2, are mapped to the corresponding regions in the depth image. This provides a powerful and extremely fast approach for establishing a real-time-capable Object Locator. In the evaluation part, the localization approach turns out to be very accurate. Nevertheless, it is dependent on the detected object itself and some additional parameters, which are analysed in this paper.

  18. Fast real-time PCR for the detection of crustacean allergen in foods.

    Science.gov (United States)

    Herrero, Beatriz; Vieites, Juan M; Espiñeira, Montserrat

    2012-02-29

    Crustaceans are one of the most common allergens causing severe food reaction. These food allergens are a health problem, and they have become very important; there are various regulations that establish that labeling must be present regarding these allergens to warn consumers. In the present work a fast real-time PCR, by a LNA probe, was developed. This allows the detection of crustaceans in all kinds of products, including processed products in which very aggressive treatments of temperature and pressure during the manufacturing process are used. This methodology provides greater sensitivity and specificity and reduces the analysis time of real-time PCR to 40 min. This methodology was further validated by means of simulating products likely to contain this allergen. For this, products present on the market were spiked with crustacean cooking water. The assay is a potential tool in issues related to the labeling of products and food security to protect the allergic consumer.

  19. Sensitive and specific detection of classical scrapie prions in the brain of goats by real-time quaking-induced conversion

    Science.gov (United States)

    The real-time quaking-induced conversion (RT-QuIC) is a rapid, specific, and sensitive prion seeding activity detection assay that uses recombinant prion protein (rPrPSen) to detect sub-infectious levels of the abnormal isoforms of the prion protein (PrPSc). Although RT-QuIC has been successfully us...

  20. FRB microstructure revealed by the real-time detection of FRB170827

    OpenAIRE

    Farah, W.; Flynn, C.; Bailes, M.; Jameson, A.; Bannister, K. W.; Barr, E. D.; Bateman, T.; Bhandari, S.; Caleb, M.; Campbell-Wilson, D.; Chang, S. -W.; Deller, A.; Green, A. J.; Hunstead, R.; Jankowski, F.

    2018-01-01

    We report a new Fast Radio Burst (FRB) discovered in real-time as part of the UTMOST project at the Molonglo Observatory Synthesis Radio Telescope (MOST). FRB170827 is the first detected with our low-latency ($< 24$ s), machine-learning-based FRB detection system. The FRB discovery was accompanied by the capture of voltage data at the native time and frequency resolution of the observing system, enabling coherent dedispersion and detailed off-line analysis, which have unveiled fine temporal a...

  1. Real-time Multiresolution Crosswalk Detection with Walk Light Recognition for the Blind

    Directory of Open Access Journals (Sweden)

    ROMIC, K.

    2018-02-01

    Full Text Available Real-time image processing and object detection techniques have a great potential to be applied in digital assistive tools for the blind and visually impaired persons. In this paper, algorithm for crosswalk detection and walk light recognition is proposed with the main aim to help blind person when crossing the road. The proposed algorithm is optimized to work in real-time on portable devices using standard cameras. Images captured by camera are processed while person is moving and decision about detected crosswalk is provided as an output along with the information about walk light if one is present. Crosswalk detection method is based on multiresolution morphological image processing, while the walk light recognition is performed by proposed 6-stage algorithm. The main contributions of this paper are accurate crosswalk detection with small processing time due to multiresolution processing and the recognition of the walk lights covering only small amount of pixels in image. The experiment is conducted using images from video sequences captured in realistic situations on crossings. The results show 98.3% correct crosswalk detections and 89.5% correct walk lights recognition with average processing speed of about 16 frames per second.

  2. Towards Real-Time Detection of Freezing of Gait Using Wavelet Transform on Wireless Accelerometer Data

    Directory of Open Access Journals (Sweden)

    Saba Rezvanian

    2016-04-01

    Full Text Available Injuries associated with fall incidences continue to pose a significant burden to persons with Parkinson’s disease (PD both in terms of human suffering and economic loss. Freezing of gait (FOG, which is one of the symptoms of PD, is a common cause of falls in this population. Although a significant amount of work has been performed to characterize/detect FOG using both qualitative and quantitative methods, there remains paucity of data regarding real-time detection of FOG, such as the requirements for minimum sensor nodes, sensor placement locations, and appropriate sampling period and update time. Here, the continuous wavelet transform (CWT is employed to define an index for correctly identifying FOG. Since the CWT method uses both time and frequency components of a waveform in comparison to other methods utilizing only the frequency component, we hypothesized that using this method could lead to a significant improvement in the accuracy of FOG detection. We tested the proposed index on the data of 10 PD patients who experience FOG. Two hundred and thirty seven (237 FOG events were identified by the physiotherapists. The results show that the index could discriminate FOG in the anterior–posterior axis better than other two axes, and is robust to the update time variability. These results suggest that real time detection of FOG may be realized by using CWT of a single shank sensor with window size of 2 s and update time of 1 s (82.1% and 77.1% for the sensitivity and specificity, respectively. Although implicated, future studies should examine the utility of this method in real-time detection of FOG.

  3. Real-time risk assessment in seismic early warning and rapid response: a feasibility study in Bishkek (Kyrgyzstan)

    Science.gov (United States)

    Picozzi, M.; Bindi, D.; Pittore, M.; Kieling, K.; Parolai, S.

    2013-04-01

    Earthquake early warning systems (EEWS) are considered to be an effective, pragmatic, and viable tool for seismic risk reduction in cities. While standard EEWS approaches focus on the real-time estimation of an earthquake's location and magnitude, innovative developments in EEWS include the capacity for the rapid assessment of damage. Clearly, for all public authorities that are engaged in coordinating emergency activities during and soon after earthquakes, real-time information about the potential damage distribution within a city is invaluable. In this work, we present a first attempt to design an early warning and rapid response procedure for real-time risk assessment. In particular, the procedure uses typical real-time information (i.e., P-wave arrival times and early waveforms) derived from a regional seismic network for locating and evaluating the size of an earthquake, information which in turn is exploited for extracting a risk map representing the potential distribution of damage from a dataset of predicted scenarios compiled for the target city. A feasibility study of the procedure is presented for the city of Bishkek, the capital of Kyrgyzstan, which is surrounded by the Kyrgyz seismic network by mimicking the ground motion associated with two historical events that occurred close to Bishkek, namely the 1911 Kemin ( M = 8.2; ±0.2) and the 1885 Belovodsk ( M = 6.9; ±0.5) earthquakes. Various methodologies from previous studies were considered when planning the implementation of the early warning and rapid response procedure for real-time risk assessment: the Satriano et al. (Bull Seismol Soc Am 98(3):1482-1494, 2008) approach to real-time earthquake location; the Caprio et al. (Geophys Res Lett 38:L02301, 2011) approach for estimating moment magnitude in real time; the EXSIM method for ground motion simulation (Motazedian and Atkinson, Bull Seismol Soc Am 95:995-1010, 2005); the Sokolov (Earthquake Spectra 161: 679-694, 2002) approach for estimating

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

    Science.gov (United States)

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

    2008-08-01

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

  5. Detection of medically important Candida species by absolute quantitation real-time polymerase chain reaction.

    Science.gov (United States)

    Than, Leslie Thian Lung; Chong, Pei Pei; Ng, Kee Peng; Seow, Heng Fong

    2015-01-01

    The number of invasive candidiasis cases has risen especially with an increase in the number of immunosuppressed and immunocom promised patients. The early detection of Candida species which is specific and sensitive is important in determining the correct administration of antifungal drugs to patients. This study aims to develop a method for the detection, identification and quantitation of medically important Candida species through quantitative polymerase chain reaction (qPCR). The isocitrate lyase (ICL) gene which is not found in mammals was chosen as the target gene of real-time PCR. Absolute quantitation of the gene copy number was achieved by constructing the plasmid containing the ICL gene which is used to generate standard curve. Twenty fungal species, two bacterial species and human DNA were tested to check the specificity of the detection method. All eight Candida species were successfully detected, identified and quantitated based on the ICL gene. A seven-log range of the gene copy number and a minimum detection limit of 10(3) copies were achieved. A one-tube absolute quantification real-time PCR that differentiates medically important Candida species via individual unique melting temperature was achieved. Analytical sensitivity and specificity were not compromised.

  6. TractorEYE: Vision-based Real-time Detection for Autonomous Vehicles in Agriculture

    DEFF Research Database (Denmark)

    Christiansen, Peter

    ) using a smaller memory footprint and 7.3-times faster processing. Low memory footprint and fast processing makes DeepAnomaly suitable for real-time applications running on an embedded GPU. FieldSAFE is a multi-modal dataset for detection of static and moving obstacles in agriculture. The dataset...... (four for rgb camera, one for thermal camera and one for a Multi-beam lidar) and fuse detection information in a common format using either 3D positions or Inverse Sensor Models. A GPU powered computational platform is able to run detection algorithms online. For the rgb camera, a deep learning...... algorithm is proposed DeepAnomaly to perform real-time anomaly detection of distant, heavy occluded and unknown obstacles in agriculture. DeepAnomaly is - compared to a state-of-the-art object detector Faster R-CNN - for an agricultural use-case able to detect humans better and at longer ranges (45-90m...

  7. Real-time Bacterial Detection by Single Cell Based Sensors UsingSynchrotron FTIR Spectromicroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Veiseh, Mandana; Veiseh, Omid; Martin, Michael C.; Bertozzi,Carolyn; Zhang, Miqin

    2005-08-10

    Microarrays of single macrophage cell based sensors weredeveloped and demonstrated for real time bacterium detection bysynchrotron FTIR microscopy. The cells were patterned on gold-SiO2substrates via a surface engineering technique by which the goldelectrodes were immobilized with fibronectin to mediate cell adhesion andthe silicon oxide background were passivated with PEG to resist proteinadsorption and cell adhesion. Cellular morphology and IR spectra ofsingle, double, and triple cells on gold electrodes exposed tolipopolysaccharide (LPS) of different concentrations were compared toreveal the detection capabilities of these biosensors. The single-cellbased sensors were found to generate the most significant IR wave numbervariation and thus provide the highest detection sensitivity. Changes inmorphology and IR spectrum for single cells exposed to LPS were found tobe time- and concentration-dependent and correlated with each other verywell. FTIR spectra from single cell arrays of gold electrodes withsurface area of 25 mu-m2, 100 mu-m2, and 400 mu-m2 were acquired usingboth synchrotron and conventional FTIR spectromicroscopes to study thesensitivity of detection. The results indicated that the developedsingle-cell platform can be used with conventional FTIRspectromicroscopy. This technique provides real-time, label-free, andrapid bacterial detection, and may allow for statistic and highthroughput analyses, and portability.

  8. A software framework for real-time multi-modal detection of microsleeps.

    Science.gov (United States)

    Knopp, Simon J; Bones, Philip J; Weddell, Stephen J; Jones, Richard D

    2017-09-01

    A software framework is described which was designed to process EEG, video of one eye, and head movement in real time, towards achieving early detection of microsleeps for prevention of fatal accidents, particularly in transport sectors. The framework is based around a pipeline structure with user-replaceable signal processing modules. This structure can encapsulate a wide variety of feature extraction and classification techniques and can be applied to detecting a variety of aspects of cognitive state. Users of the framework can implement signal processing plugins in C++ or Python. The framework also provides a graphical user interface and the ability to save and load data to and from arbitrary file formats. Two small studies are reported which demonstrate the capabilities of the framework in typical applications: monitoring eye closure and detecting simulated microsleeps. While specifically designed for microsleep detection/prediction, the software framework can be just as appropriately applied to (i) other measures of cognitive state and (ii) development of biomedical instruments for multi-modal real-time physiological monitoring and event detection in intensive care, anaesthesiology, cardiology, neurosurgery, etc. The software framework has been made freely available for researchers to use and modify under an open source licence.

  9. Visual detectability of elastic contrast in real-time ultrasound images

    Science.gov (United States)

    Miller, Naomi R.; Bamber, Jeffery C.; Doyley, Marvin M.; Leach, Martin O.

    1997-04-01

    Elasticity imaging (EI) has recently been proposed as a technique for imaging the mechanical properties of soft tissue. However, dynamic features, known as compressibility and mobility, are already employed to distinguish between different tissue types in ultrasound breast examination. This method, which involves the subjective interpretation of tissue motion seen in real-time B-mode images during palpation, is hereafter referred to as differential motion imaging (DMI). The purpose of this study was to develop the methodology required to perform a series of perception experiments to measure elastic lesion detectability by means of DMI and to obtain preliminary results for elastic contrast thresholds for different lesion sizes. Simulated sequences of real-time B-scans of tissue moving in response to an applied force were generated. A two-alternative forced choice (2-AFC) experiment was conducted and the measured contrast thresholds were compared with published results for lesions detected by EI. Although the trained observer was found to be quite skilled at the task of differential motion perception, it would appear that lesion detectability is improved when motion information is detected by computer processing and converted to gray scale before presentation to the observer. In particular, for lesions containing fewer than eight speckle cells, a signal detection rate of 100% could not be achieved even when the elastic contrast was very high.

  10. Real-time DNA Amplification and Detection System Based on a CMOS Image Sensor.

    Science.gov (United States)

    Wang, Tiantian; Devadhasan, Jasmine Pramila; Lee, Do Young; Kim, Sanghyo

    2016-01-01

    In the present study, we developed a polypropylene well-integrated complementary metal oxide semiconductor (CMOS) platform to perform the loop mediated isothermal amplification (LAMP) technique for real-time DNA amplification and detection simultaneously. An amplification-coupled detection system directly measures the photon number changes based on the generation of magnesium pyrophosphate and color changes. The photon number decreases during the amplification process. The CMOS image sensor observes the photons and converts into digital units with the aid of an analog-to-digital converter (ADC). In addition, UV-spectral studies, optical color intensity detection, pH analysis, and electrophoresis detection were carried out to prove the efficiency of the CMOS sensor based the LAMP system. Moreover, Clostridium perfringens was utilized as proof-of-concept detection for the new system. We anticipate that this CMOS image sensor-based LAMP method will enable the creation of cost-effective, label-free, optical, real-time and portable molecular diagnostic devices.

  11. Single reaction, real time RT-PCR detection of all known avian and human metapneumoviruses.

    Science.gov (United States)

    Lemaitre, E; Allée, C; Vabret, A; Eterradossi, N; Brown, P A

    2018-01-01

    Current molecular methods for the detection of avian and human metapneumovirus (AMPV, HMPV) are specifically targeted towards each virus species or individual subgroups of these. Here a broad range SYBR Green I real time RT-PCR was developed which amplified a highly conserved fragment of sequence in the N open reading frame. This method was sufficiently efficient and specific in detecting all MPVs. Its validation according to the NF U47-600 norm for the four AMPV subgroups estimated low limits of detection between 1000 and 10copies/μL, similar with detection levels described previously for real time RT-PCRs targeting specific subgroups. RNA viruses present a challenge for the design of durable molecular diagnostic test due to the rate of change in their genome sequences which can vary substantially in different areas and over time. The fact that the regions of sequence for primer hybridization in the described method have remained sufficiently conserved since the AMPV and HMPV diverged, should give the best chance of continued detection of current subgroups and of potential unknown or future emerging MPV strains. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Roman Wölfel

    2012-08-01

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

  13. Algorithm for real-time detection of signal patterns using phase synchrony: an application to an electrode array

    Science.gov (United States)

    Sadeghi, Saman; MacKay, William A.; van Dam, R. Michael; Thompson, Michael

    2011-02-01

    Real-time analysis of multi-channel spatio-temporal sensor data presents a considerable technical challenge for a number of applications. For example, in brain-computer interfaces, signal patterns originating on a time-dependent basis from an array of electrodes on the scalp (i.e. electroencephalography) must be analyzed in real time to recognize mental states and translate these to commands which control operations in a machine. In this paper we describe a new technique for recognition of spatio-temporal patterns based on performing online discrimination of time-resolved events through the use of correlation of phase dynamics between various channels in a multi-channel system. The algorithm extracts unique sensor signature patterns associated with each event during a training period and ranks importance of sensor pairs in order to distinguish between time-resolved stimuli to which the system may be exposed during real-time operation. We apply the algorithm to electroencephalographic signals obtained from subjects tested in the neurophysiology laboratories at the University of Toronto. The extension of this algorithm for rapid detection of patterns in other sensing applications, including chemical identification via chemical or bio-chemical sensor arrays, is also discussed.

  14. A Portable Array-Type Optical Fiber Sensing Instrument for Real-Time Gas Detection

    Directory of Open Access Journals (Sweden)

    San-Shan Hung

    2016-12-01

    Full Text Available A novel optical fiber array-type of sensing instrument with temperature compensation for real-time detection was developed to measure oxygen, carbon dioxide, and ammonia simultaneously. The proposed instrument is multi-sensing array integrated with real-time measurement module for portable applications. The sensing optical fibers were etched and polished before coating to increase sensitivities. The ammonia and temperature sensors were each composed of a dye-coated single-mode fiber with constructing a fiber Bragg grating and a long-period filter grating for detecting light intensity. Both carbon dioxide and oxygen sensing structures use multimode fibers where 1-hydroxy-3,6,8-pyrene trisulfonic acid trisodium salt is coated for carbon dioxide sensing and Tris(2,2′-bipyridyl dichlororuthenium(II hexahydrate and Tris(bipyridineruthenium(II chloride are coated for oxygen sensing. Gas-induced fluorescent light intensity variation was applied to detect gas concentration. The portable gas sensing array was set up by integrating with photo-electronic measurement modules and a human-machine interface to detect gases in real time. The measured data have been processed using piecewise-linear method. The sensitivity of the oxygen sensor were 1.54%/V and 9.62%/V for concentrations less than 1.5% and for concentrations between 1.5% and 6%, respectively. The sensitivity of the carbon dioxide sensor were 8.33%/V and 9.62%/V for concentrations less than 2% and for concentrations between 2% and 5%, respectively. For the ammonia sensor, the sensitivity was 27.78%/V, while ammonia concentration was less than 2%.

  15. A Portable Array-Type Optical Fiber Sensing Instrument for Real-Time Gas Detection.

    Science.gov (United States)

    Hung, San-Shan; Chang, Hsing-Cheng; Chang, I-Nan

    2016-12-08

    A novel optical fiber array-type of sensing instrument with temperature compensation for real-time detection was developed to measure oxygen, carbon dioxide, and ammonia simultaneously. The proposed instrument is multi-sensing array integrated with real-time measurement module for portable applications. The sensing optical fibers were etched and polished before coating to increase sensitivities. The ammonia and temperature sensors were each composed of a dye-coated single-mode fiber with constructing a fiber Bragg grating and a long-period filter grating for detecting light intensity. Both carbon dioxide and oxygen sensing structures use multimode fibers where 1-hydroxy-3,6,8-pyrene trisulfonic acid trisodium salt is coated for carbon dioxide sensing and Tris(2,2'-bipyridyl) dichlororuthenium(II) hexahydrate and Tris(bipyridine)ruthenium(II) chloride are coated for oxygen sensing. Gas-induced fluorescent light intensity variation was applied to detect gas concentration. The portable gas sensing array was set up by integrating with photo-electronic measurement modules and a human-machine interface to detect gases in real time. The measured data have been processed using piecewise-linear method. The sensitivity of the oxygen sensor were 1.54%/V and 9.62%/V for concentrations less than 1.5% and for concentrations between 1.5% and 6%, respectively. The sensitivity of the carbon dioxide sensor were 8.33%/V and 9.62%/V for concentrations less than 2% and for concentrations between 2% and 5%, respectively. For the ammonia sensor, the sensitivity was 27.78%/V, while ammonia concentration was less than 2%.

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

    Science.gov (United States)

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

    2014-05-30

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

  17. Detection of intestinal protozoa in paediatric patients with gastrointestinal symptoms by multiplex real-time PCR.

    Science.gov (United States)

    Maas, L; Dorigo-Zetsma, J W; de Groot, C J; Bouter, S; Plötz, F B; van Ewijk, B E

    2014-06-01

    The performance of a multiplex real-time PCR for the detection of Blastocystis, Dientamoeba fragilis, Giardia lamblia, Cryptosporidium species and Entamoeba species in faecal samples was evaluated in an observational prospective study. Paediatric patients (0-18 years) presenting with gastrointestinal symptoms and suspected of having enteroparasitic disease were included. A questionnaire on gastrointestinal symptoms and the chosen treatment was completed at the start of the study and after 6 weeks. Of 163 paediatric patients (mean age, 7.8 years), 114 (70%) had a PCR-positive faecal sample. D. fragilis was detected most frequently, in 101 patients, followed by Blastocystis in 49. In faecal samples of 47 patients, more than one protozoan was detected, mainly the combination of D. fragilis and Blastocystis. Reported gastrointestinal symptoms were abdominal pain (78%), nausea (30%), and altered bowel habits (28%). Eighty-nine of the PCR-positive patients were treated with antibiotics. A significant reduction in abdominal pain was observed both in treated and in untreated patients. This study demonstrated that multiplex real-time PCR detects a high percentage of intestinal protozoa in paediatric patients with gastrointestinal symptoms. However, interpretation and determination of the clinical relevance of a positive PCR result in this population are still difficult. © 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.

  18. EVALUATION OF A RAPID, QUANTITATIVE REAL-TIME PCR METHOD FOR ENUMERATION OF PATHOGENIC CANDIDA CELLS IN WATER

    Science.gov (United States)

    Quantitative Real-Time PCR (QRT-PCR) technology, incorporating fluorigenic 5' nuclease (TaqMan?) chemistry, was developed for the specific detection and quantification of six pathogenic species of Candida (C. albicans, C. tropicalis, C. krusei, C. parapsilosis, C. glabrata and C....

  19. Embedded and real-time vehicle detection system for challenging on-road scenes

    Science.gov (United States)

    Gu, Qin; Yang, Jianyu; Kong, Lingjiang; Yan, Wei Qi; Klette, Reinhard

    2017-06-01

    Vehicle detection is an important topic for advanced driver-assistance systems. This paper proposes an adaptive approach for an embedded system by focusing on monocular vehicle detection in real time, also aiming at being accurate under challenging conditions. Scene classification is accomplished by using a simplified convolution neural network with hypothesis generation by SoftMax regression. The output is consequently taken into account to optimize detection parameters for hypothesis generation and testing. Thus, we offer a sample-reorganization mechanism to improve the performance of vehicle hypothesis verification. A hypothesis leap mechanism is in use to improve the operating efficiency of the on-board system. A practical on-road test is employed to verify vehicle detection (i.e., accuracy) and also the performance of the designed on-board system regarding speed.

  20. Highly sensitive real-time PCR for specific detection and quantification of Coxiella burnetii

    Directory of Open Access Journals (Sweden)

    Linke Sonja

    2006-01-01

    Full Text Available Abstract Background Coxiella burnetii, the bacterium causing Q fever, is an obligate intracellular biosafety level 3 agent. Detection and quantification of these bacteria with conventional methods is time consuming and dangerous. During the last years, several PCR based diagnostic assays were developed to detect C. burnetii DNA in cell cultures and clinical samples. We developed and evaluated TaqMan-based real-time PCR assays that targeted the singular icd (isocitrate dehydrogenase gene and the transposase of the IS1111a element present in multiple copies in the C. burnetii genome. Results To evaluate the precision of the icd and IS1111 real-time PCR assays, we performed different PCR runs with independent DNA dilutions of the C. burnetii Nine Mile RSA493 strain. The results showed very low variability, indicating efficient reproducibility of both assays. Using probit analysis, we determined that the minimal number of genome equivalents per reaction that could be detected with a 95% probability was 10 for the icd marker and 6.5 for the IS marker. Plasmid standards with cloned icd and IS1111 fragments were used to establish standard curves which were linear over a range from 10 to 107 starting plasmid copy numbers. We were able to quantify cell numbers of a diluted, heat-inactivated Coxiella isolate with a detection limit of 17 C. burnetii particles per reaction. Real-time PCR targeting both markers was performed with DNA of 75 different C. burnetii isolates originating from all over the world. Using this approach, the number of IS1111 elements in the genome of the Nine Mile strain was determined to be 23, close to 20, the number revealed by genome sequencing. In other isolates, the number of IS1111 elements varied widely (between seven and 110 and seemed to be very high in some isolates. Conclusion We validated TaqMan-based real-time PCR assays targeting the icd and IS1111 markers of C. burnetii. The assays were shown to be specific, highly

  1. Real-time flight conflict detection and release based on Multi-Agent system

    Science.gov (United States)

    Zhang, Yifan; Zhang, Ming; Yu, Jue

    2018-01-01

    This paper defines two-aircrafts, multi-aircrafts and fleet conflict mode, sets up space-time conflict reservation on the basis of safety interval and conflict warning time in three-dimension. Detect real-time flight conflicts combined with predicted flight trajectory of other aircrafts in the same airspace, and put forward rescue resolutions for the three modes respectively. When accorded with the flight conflict conditions, determine the conflict situation, and enter the corresponding conflict resolution procedures, so as to avoid the conflict independently, as well as ensure the flight safety of aimed aircraft. Lastly, the correctness of model is verified with numerical simulation comparison.

  2. Detection, quantification and genotyping of Herpes Simplex Virus in cervicovaginal secretions by real-time PCR: a cross sectional survey

    Directory of Open Access Journals (Sweden)

    Natividad-Sancho Angels

    2005-08-01

    Full Text Available Abstract Background Herpes Simplex Virus (HSV Genital Ulcer Disease (GUD is an important public health problem, whose interaction with HIV results in mutually enhancing epidemics. Conventional methods for detecting HSV tend to be slow and insensitive. We designed a rapid PCR-based assay to quantify and type HSV in cervicovaginal lavage (CVL fluid of subjects attending a Genito-Urinary Medicine (GUM clinic. Vaginal swabs, CVL fluid and venous blood were collected. Quantitative detection of HSV was conducted using real time PCR with HSV specific primers and SYBR Green I. Fluorogenic TaqMan Minor Groove Binder (MGB probes designed around a single base mismatch in the HSV DNA polymerase I gene were used to type HSV in a separate reaction. The Kalon test was used to detect anti-HSV-2 IgG antibodies in serum. Testing for HIV, other Sexually Transmitted Infections (STI and related infections was based on standard clinical and laboratory methods. Results Seventy consecutive GUM clinic attendees were studied. Twenty-seven subjects (39% had detectable HSV DNA in CVL fluid; HSV-2 alone was detected in 19 (70% subjects, HSV-1 alone was detected in 4 (15% subjects and both HSV types were detected in 4 (15% subjects. Eleven out of 27 subjects (41% with anti-HSV-2 IgG had detectable HSV-2 DNA in CVL fluid. Seven subjects (10% were HIV-positive. Three of seven (43% HIV-infected subjects and two of five subjects with GUD (40% were secreting HSV-2. None of the subjects in whom HSV-1 was detected had GUD. Conclusion Quantitative real-time PCR and Taqman MGB probes specific for HSV-1 or -2 were used to develop an assay for quantification and typing of HSV. The majority of subjects in which HSV was detected had low levels of CVL fluid HSV, with no detectable HSV-2 antibodies and were asymptomatic.

  3. Real-time prostate-specific antigen detection with prostate-specific antigen imprinted capacitive biosensors

    Energy Technology Data Exchange (ETDEWEB)

    Ertürk, Gizem [Department of Biotechnology, Lund University, Lund (Sweden); Department of Biology, Hacettepe University, Ankara (Turkey); Hedström, Martin [Department of Biotechnology, Lund University, Lund (Sweden); CapSenze HB, Medicon Village, SE-223 63 Lund (Sweden); Tümer, M. Aşkın [Department of Biology, Hacettepe University, Ankara (Turkey); Denizli, Adil [Department of Chemistry, Hacettepe University, Ankara (Turkey); Mattiasson, Bo, E-mail: Bo.Mattiasson@biotek.lu.se [Department of Biotechnology, Lund University, Lund (Sweden); CapSenze HB, Medicon Village, SE-223 63 Lund (Sweden)

    2015-09-03

    Prostate specific antigen (PSA) is a valuable biomarker for early detection of prostate cancer, the third most common cancer in men. Ultrasensitive detection of PSA is crucial to screen the prostate cancer in an early stage and to detect the recurrence of the disease after treatment. In this report, microcontact-PSA imprinted (PSA-MIP) capacitive biosensor chip was developed for real-time, highly sensitive and selective detection of PSA. PSA-MIP electrodes were prepared in the presence of methacrylic acid (MAA) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as the cross-linker via UV polymerization. Immobilized Anti-PSA antibodies on electrodes (Anti-PSA) for capacitance measurements were also prepared to compare the detection performances of both methods. The electrodes were characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM) and cyclic voltammetry (CV) and real-time PSA detection was performed with standard PSA solutions in the concentration range of 10 fg mL{sup −1}–100 ng mL{sup −1}. The detection limits were found as 8.0 × 10{sup −5} ng mL{sup −1} (16 × 10{sup −17} M) and 6.0 × 10{sup −4} ng mL{sup −1} (12 × 10{sup −16} M) for PSA-MIP and Anti-PSA electrodes, respectively. Selectivity studies were performed against HSA and IgG and selectivity coefficients were calculated. PSA detection was also carried out from diluted human serum samples and finally, reproducibility of the electrodes was tested. The results are promising and show that when the sensitivity of the capacitive system is combined with the selectivity and reproducibility of the microcontact-imprinting procedure, the resulting system might be used successfully for real-time detection of various analytes even in very low concentrations. - Highlights: • Microcontact imprinting method was used for preparing the sensor chip for capacitive biosensing. • High sensitivity was obtained. • Good selectivity was

  4. Real-time prostate-specific antigen detection with prostate-specific antigen imprinted capacitive biosensors

    International Nuclear Information System (INIS)

    Ertürk, Gizem; Hedström, Martin; Tümer, M. Aşkın; Denizli, Adil; Mattiasson, Bo

    2015-01-01

    Prostate specific antigen (PSA) is a valuable biomarker for early detection of prostate cancer, the third most common cancer in men. Ultrasensitive detection of PSA is crucial to screen the prostate cancer in an early stage and to detect the recurrence of the disease after treatment. In this report, microcontact-PSA imprinted (PSA-MIP) capacitive biosensor chip was developed for real-time, highly sensitive and selective detection of PSA. PSA-MIP electrodes were prepared in the presence of methacrylic acid (MAA) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as the cross-linker via UV polymerization. Immobilized Anti-PSA antibodies on electrodes (Anti-PSA) for capacitance measurements were also prepared to compare the detection performances of both methods. The electrodes were characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM) and cyclic voltammetry (CV) and real-time PSA detection was performed with standard PSA solutions in the concentration range of 10 fg mL"−"1–100 ng mL"−"1. The detection limits were found as 8.0 × 10"−"5 ng mL"−"1 (16 × 10"−"1"7 M) and 6.0 × 10"−"4 ng mL"−"1 (12 × 10"−"1"6 M) for PSA-MIP and Anti-PSA electrodes, respectively. Selectivity studies were performed against HSA and IgG and selectivity coefficients were calculated. PSA detection was also carried out from diluted human serum samples and finally, reproducibility of the electrodes was tested. The results are promising and show that when the sensitivity of the capacitive system is combined with the selectivity and reproducibility of the microcontact-imprinting procedure, the resulting system might be used successfully for real-time detection of various analytes even in very low concentrations. - Highlights: • Microcontact imprinting method was used for preparing the sensor chip for capacitive biosensing. • High sensitivity was obtained. • Good selectivity was demonstrated. • Stability of

  5. Detection of diarrheagenic Escherichia coli by use of melting-curve analysis and real-time multiplex PCR.

    Science.gov (United States)

    Guion, Chase E; Ochoa, Theresa J; Walker, Christopher M; Barletta, Francesca; Cleary, Thomas G

    2008-05-01

    Diarrheagenic Escherichia coli strains are important causes of diarrhea in children from the developing world and are now being recognized as emerging enteropathogens in the developed world. Current methods of detection are too expensive and labor-intensive for routine detection of these organisms to be practical. We developed a real-time fluorescence-based multiplex PCR for the detection of all six of the currently recognized classes of diarrheagenic E. coli. The primers were designed to specifically amplify eight different virulence genes in the same reaction: aggR for enteroaggregative E. coli, stIa/stIb and lt for enterotoxigenic E. coli, eaeA for enteropathogenic E. coli and Shiga toxin-producing E. coli (STEC), stx(1) and stx(2) for STEC, ipaH for enteroinvasive E. coli, and daaD for diffusely adherent E. coli (DAEC). Eighty-nine of ninety diarrheagenic E. coli and 36/36 nonpathogenic E. coli strains were correctly identified using this approach (specificity, 1.00; sensitivity, 0.99). The single false negative was a DAEC strain. The total time between preparation of DNA from E. coli colonies on agar plates and completion of PCR and melting-curve analysis was less than 90 min. The cost of materials was low. Melting-point analysis of real-time multiplex PCR is a rapid, sensitive, specific, and inexpensive method for detection of diarrheagenic E. coli.

  6. A real-time reverse transcriptase polymerase chain reaction for detection and quantification of Vesiculovirus

    Directory of Open Access Journals (Sweden)

    Aline Lavado Tolardo

    2016-06-01

    Full Text Available Vesiculoviruses (VSV are zoonotic viruses that cause vesicular stomatitis disease in cattle, horses and pigs, as well as sporadic human cases of acute febrile illness. Therefore, diagnosis of VSV infections by reliable laboratory techniques is important to allow a proper case management and implementation of strategies for the containment of virus spread. We show here a sensitive and reproducible real-time reverse transcriptase polymerase chain reaction (RT-PCR for detection and quantification of VSV. The assay was evaluated with arthropods and serum samples obtained from horses, cattle and patients with acute febrile disease. The real-time RT-PCR amplified the Piry, Carajas, Alagoas and Indiana Vesiculovirus at a melting temperature 81.02 ± 0.8ºC, and the sensitivity of assay was estimated in 10 RNA copies/mL to the Piry Vesiculovirus. The viral genome has been detected in samples of horses and cattle, but not detected in human sera or arthropods. Thus, this assay allows a preliminary differential diagnosis of VSV infections.

  7. Real-time detection of transient cardiac ischemic episodes from ECG signals

    International Nuclear Information System (INIS)

    Dranca, L; Goñi, A; Illarramendi, A

    2009-01-01

    We propose a new algorithm to detect and classify transient cardiac ischemia episodes, designed with the goal of providing a real-time execution without penalizing the classifier accuracy much. The algorithm is based on a novel mixture of time-domain analysis and machine learning techniques, specifically bagging of decision trees, and it has been developed using a well-recognized and freely distributed database, namely the long-term ST database. The ST episode detection sensitivity/positive predictivity using the annotation protocol A for this database is 68.26%/74.91%. The sensitivity result increases until 93.97% for the most dangerous episodes in terms of duration and magnitude (annotated according to protocol C). The test of the algorithm over the freely distributed part of the European Society of Cardiology database has shown results of sensitivity and positive predictivity of 83.33% and 77.31%, respectively. Those results are close to the results obtained by related works that present approaches to detect ischemia episodes off-line, which is remarkable if we take into account that in our real-time approach, less information is available during the classification process

  8. Real time failure detection in unreinforced cementitious composites with triboluminescent sensor

    International Nuclear Information System (INIS)

    Olawale, David O.; Kliewer, Kaitlyn; Okoye, Annuli; Dickens, Tarik J.; Uddin, Mohammed J.; Okoli, Okenwa I.

    2014-01-01

    The in-situ triboluminescent optical fiber (ITOF) sensor has an integrated sensing and transmission component that converts the energy from damage events like impacts and crack propagation into optical signals that are indicative of the magnitude of damage in composite structures like concrete bridges. Utilizing the triboluminescence (TL) property of ZnS:Mn, the ITOF sensor has been successfully integrated into unreinforced cementitious composite beams to create multifunctional smart structures with in-situ failure detection capabilities. The fabricated beams were tested under flexural loading, and real time failure detection was made by monitoring the TL signals generated by the integrated ITOF sensor. Tested beam samples emitted distinctive TL signals at the instance of failure. In addition, we report herein a new and promising approach to damage characterization using TL emission profiles. Analysis of TL emission profiles indicates that the ITOF sensor responds to crack propagation through the beam even when not in contact with the crack. Scanning electron microscopy analysis indicated that fracto-triboluminescence was responsible for the TL signals observed at the instance of beam failure. -- Highlights: • Developed a new approach to triboluminescence (TL)-based sensing with ZnS:Mn. • Damage-induced excitation of ZnS:Mn enabled real time damage detection in composite. • Based on sensor position, correlation exists between TL signal and failure stress. • Introduced a new approach to damage characterization with TL profile analysis

  9. Novel methods of cytokine detection: Real-time PCR, ELISPOT, and intracellular cytokine staining

    Directory of Open Access Journals (Sweden)

    Eliza Turlej

    2009-05-01

    Full Text Available Cytokines are small hormone-like proteins that play important roles in immune system control. Cytokines regulate the proliferation and differentiation of cells and hematopoiesis and act as mediators in the inflammatory reaction. Changes in cytokine levels are found in many diseases, such as sepsis, bowel inflammatory disease, autoimmune diseases, as well as graft-versus-host disease. Cytokines levels can be detected using in vivo, in vitro, and ex vivo techniques. The level of cytokine produced can be measured by immunoenzymatic test (ELISA in supernatant after cell culture with the addition of stimulant and in plasma by techniques that measure the level of cytokine secretion in cells (e.g. immunohistochemical staining, ELISPOT, and intracellular cytokine staining, and by molecular biological methods (RPA, real-time PCR, in situ hybridization, and Northern blot. Detection of cytokine mRNA in tissues is useful in the direct determination of heterogenic populations of cytokine-producing cells. Nowadays the most frequently used methods for measuring cytokine level are ELISPOT, intracellular cytokine staining with flow cytometry detection, and real-time PCR. These methods have an important clinical role in vaccine efficacy, in viral, bacterial, and verminous diagnostics, and in determining the efficacy of cancer treatment.

  10. A Real-time License Plate Detection System for Parking Access

    Directory of Open Access Journals (Sweden)

    Roenadi Koesdijarto

    2010-08-01

    Full Text Available The automatic and real-time license plate detection system can be used as an access control entry of vehicles into the parking area. The problem is how to recognize the vehicles that will go into the parking lot and how to recognize various types of license plates in various light conditions quickly and accurately. In this research, the prototype was developed with a detection system to recognize the vehicles that will enter the parking area, and a license plate recognition system. In the license plate recognition system, the Fourier transform and Hidden Markov model method have proposed to detect location of license plate and as characters segmentation to recognize Indonesia license plates. The research results have shown that the developed prototype system has successfully recognized all Indonesia license plates in several of light condition and camera position. The percentage of plate recognition in the real-time experiment is 84.38%, and the average execution time for all recognition process is 5.834 second.

  11. Dynamic modelling and real-time leak detection for NGL pipelines

    Energy Technology Data Exchange (ETDEWEB)

    Young, B.R.; Svrcek, W.Y. [Calgary Univ., AB (Canada). Dept. of Chemical and Petroleum Engineering; Cooke, J.G.; Daye, R.E. [Rangeland Engineering Ltd., Calgary, AB (Canada)

    2004-07-01

    This paper presented newly developed steady-state and dynamic models commissioned for natural gas liquids (NGL) pipelines near Empress, Alberta. The work demonstrates a unique university-industry collaboration for solving the challenge of reliable pipeline leak detection. The flexible, custom real-time leak detection system was tested on the dynamic simulation. It was successfully used to replace a volume balance system for NGL pipelines at Empress in March 2003. A custom pipeline monitoring system was also developed to integrate with the existing pipeline supervisory control and data acquisition (SCADA) system. Simulation results enabled a change in the control scheme of the pipelines that resulted in less transient operation. The premise of the leak detection system is a rigorous thermodynamics and dynamic mass balance calculation based on real-time information from field flow, pressure and temperature sensors. The application of the system makes it possible to minimize or eliminate false or nuisance alarms, which is critical to the confidence of the monitoring system. The volumetric and mass imbalance formulae permit the system to cross check the calculation and then make important decisions regarding the sounding of alarms. The custom solution offers flexibility for use in a wide variety of conditions and applications. In addition, it is cost effective and locally supported. 8 refs., 4 tabs., 5 figs.

  12. Multiplex real-time PCR SYBR Green for detection and typing of group III Clostridium botulinum.

    Science.gov (United States)

    Anniballi, Fabrizio; Auricchio, Bruna; Delibato, Elisabetta; Antonacci, Monia; De Medici, Dario; Fenicia, Lucia

    2012-01-27

    Clostridium botulinum type C and type D belonging to the group III organisms, are mainly responsible for animal botulism outbreaks. Clinical signs alone are often insufficient to make a diagnosis of botulism and a laboratory confirmation is required. Laboratory confirmation can be performed by demonstrating the presence of botulinum neurotoxins in serum, gastrointestinal contents, liver, wound of sick or dead animals, or by demonstrating the presence of C. botulinum in gastrointestinal contents, liver, and wound. Demonstration of spores in gastrointestinal contents or tissue of animals with clinical signs indicative of botulism reinforces the clinical diagnosis. With the aim of detecting and typing C. botulinum group III organisms, a multiplex real-time PCR SYBR Green was developed and in-house validated. Selectivity, limit of detection, relative accuracy, relative specificity, relative sensitivity, and repeatability of the method were investigated. The multiplex real-time PCR SYBR green used showed a 100% selectivity, 100% relative accuracy, 100% relative specificity, 100% relative sensitivity and a limit of detection of 277 and 580 DNA copies for C. botulinum type C and C. botulinum type D, respectively. The method reported here represents a suitable tool for laboratory diagnosis of type C and D botulism and for testing a large number of samples collected during the animal botulism surveillance and prevention activities. Copyright © 2011 Elsevier B.V. All rights reserved.

  13. Matching-range-constrained real-time loop closure detection with CNNs features.

    Science.gov (United States)

    Bai, Dongdong; Wang, Chaoqun; Zhang, Bo; Yi, Xiaodong; Tang, Yuhua

    2016-01-01

    The loop closure detection (LCD) is an essential part of visual simultaneous localization and mapping systems (SLAM). LCD is capable of identifying and compensating the accumulation drift of localization algorithms to produce an consistent map if the loops are checked correctly. Deep convolutional neural networks (CNNs) have outperformed state-of-the-art solutions that use traditional hand-crafted features in many computer vision and pattern recognition applications. After the great success of CNNs, there has been much interest in applying CNNs features to robotic fields such as visual LCD. Some researchers focus on using a pre-trained CNNs model as a method of generating an image representation appropriate for visual loop closure detection in SLAM. However, there are many fundamental differences and challenges involved in character between simple computer vision applications and robotic applications. Firstly, the adjacent images in the dataset of loop closure detection might have more resemblance than the images that form the loop closure. Secondly, real-time performance is one of the most critical demands for robots. In this paper, we focus on making use of the feature generated by CNNs layers to implement LCD in real environment. In order to address the above challenges, we explicitly provide a value to limit the matching range of images to solve the first problem; meanwhile we get better results than state-of-the-art methods and improve the real-time performance using an efficient feature compression method.

  14. A Precise and Real-Time Loop-closure Detection for SLAM Using the RSOM Tree

    Directory of Open Access Journals (Sweden)

    Siyang Song

    2015-06-01

    Full Text Available In robotic applications of visual simultaneous localization and mapping (SLAM techniques, loop-closure detection detects whether or not a current location has previously been visited. We present an online and incremental approach to detect loops when images come from an already visited scene and learn new information from the environment. Instead of utilizing a bag-of-words model, the attributed graph model is applied to represent images and measure the similarity between pairs of images in our method. In order to position a camera in visual environments in real-time, the method demands retrieval of images from the database through a clustering tree that we call RSOM (recursive self-organizing feature map. As long as the match is found between the current graph and several graphs in the database, a threshold will be chosen to judge whether loop-closure is accepted or rejected. The results demonstrate the method's accuracy and real-time performance by testing several videos collected from a digital camera fixed on vehicles in indoor and outdoor environments.

  15. Detection of tumor markers in prostate cancer and comparison of sensitivity between real time and nested PCR.

    Science.gov (United States)

    Matsuoka, Takayuki; Shigemura, Katsumi; Yamamichi, Fukashi; Fujisawa, Masato; Kawabata, Masato; Shirakawa, Toshiro

    2012-06-27

    The objective of this study is to investigate and compare the sensitivity in conventional PCR, quantitative real time PCR, nested PCR and western blots for detection of prostate cancer tumor markers using prostate cancer (PCa) cells. We performed conventional PCR, quantitative real time PCR, nested PCR, and western blots using 5 kinds of PCa cells. Prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), and androgen receptor (AR) were compared for their detection sensitivity by real time PCR and nested PCR. In real time PCR, there was a significant correlation between cell number and the RNA concentration obtained (R(2)=0.9944) for PSA, PSMA, and AR. We found it possible to detect these markers from a single LNCaP cell in both real time and nested PCR. By comparison, nested PCR reached a linear curve in fewer PCR cycles than real time PCR, suggesting that nested PCR may offer PCR results more quickly than real time PCR. In conclusion, nested PCR may offer tumor maker detection in PCa cells more quickly (with fewer PCR cycles) with the same high sensitivity as real time PCR. Further study is necessary to establish and evaluate the best tool for PCa tumor marker detection.

  16. Real-time community detection in full social networks on a laptop

    Science.gov (United States)

    Chamberlain, Benjamin Paul; Levy-Kramer, Josh; Humby, Clive

    2018-01-01

    For a broad range of research and practical applications it is important to understand the allegiances, communities and structure of key players in society. One promising direction towards extracting this information is to exploit the rich relational data in digital social networks (the social graph). As global social networks (e.g., Facebook and Twitter) are very large, most approaches make use of distributed computing systems for this purpose. Distributing graph processing requires solving many difficult engineering problems, which has lead some researchers to look at single-machine solutions that are faster and easier to maintain. In this article, we present an approach for analyzing full social networks on a standard laptop, allowing for interactive exploration of the communities in the locality of a set of user specified query vertices. The key idea is that the aggregate actions of large numbers of users can be compressed into a data structure that encapsulates the edge weights between vertices in a derived graph. Local communities can be constructed by selecting vertices that are connected to the query vertices with high edge weights in the derived graph. This compression is robust to noise and allows for interactive queries of local communities in real-time, which we define to be less than the average human reaction time of 0.25s. We achieve single-machine real-time performance by compressing the neighborhood of each vertex using minhash signatures and facilitate rapid queries through Locality Sensitive Hashing. These techniques reduce query times from hours using industrial desktop machines operating on the full graph to milliseconds on standard laptops. Our method allows exploration of strongly associated regions (i.e., communities) of large graphs in real-time on a laptop. It has been deployed in software that is actively used by social network analysts and offers another channel for media owners to monetize their data, helping them to continue to provide

  17. Real-time community detection in full social networks on a laptop.

    Directory of Open Access Journals (Sweden)

    Benjamin Paul Chamberlain

    Full Text Available For a broad range of research and practical applications it is important to understand the allegiances, communities and structure of key players in society. One promising direction towards extracting this information is to exploit the rich relational data in digital social networks (the social graph. As global social networks (e.g., Facebook and Twitter are very large, most approaches make use of distributed computing systems for this purpose. Distributing graph processing requires solving many difficult engineering problems, which has lead some researchers to look at single-machine solutions that are faster and easier to maintain. In this article, we present an approach for analyzing full social networks on a standard laptop, allowing for interactive exploration of the communities in the locality of a set of user specified query vertices. The key idea is that the aggregate actions of large numbers of users can be compressed into a data structure that encapsulates the edge weights between vertices in a derived graph. Local communities can be constructed by selecting vertices that are connected to the query vertices with high edge weights in the derived graph. This compression is robust to noise and allows for interactive queries of local communities in real-time, which we define to be less than the average human reaction time of 0.25s. We achieve single-machine real-time performance by compressing the neighborhood of each vertex using minhash signatures and facilitate rapid queries through Locality Sensitive Hashing. These techniques reduce query times from hours using industrial desktop machines operating on the full graph to milliseconds on standard laptops. Our method allows exploration of strongly associated regions (i.e., communities of large graphs in real-time on a laptop. It has been deployed in software that is actively used by social network analysts and offers another channel for media owners to monetize their data, helping them to

  18. Real-time community detection in full social networks on a laptop.

    Science.gov (United States)

    Chamberlain, Benjamin Paul; Levy-Kramer, Josh; Humby, Clive; Deisenroth, Marc Peter

    2018-01-01

    For a broad range of research and practical applications it is important to understand the allegiances, communities and structure of key players in society. One promising direction towards extracting this information is to exploit the rich relational data in digital social networks (the social graph). As global social networks (e.g., Facebook and Twitter) are very large, most approaches make use of distributed computing systems for this purpose. Distributing graph processing requires solving many difficult engineering problems, which has lead some researchers to look at single-machine solutions that are faster and easier to maintain. In this article, we present an approach for analyzing full social networks on a standard laptop, allowing for interactive exploration of the communities in the locality of a set of user specified query vertices. The key idea is that the aggregate actions of large numbers of users can be compressed into a data structure that encapsulates the edge weights between vertices in a derived graph. Local communities can be constructed by selecting vertices that are connected to the query vertices with high edge weights in the derived graph. This compression is robust to noise and allows for interactive queries of local communities in real-time, which we define to be less than the average human reaction time of 0.25s. We achieve single-machine real-time performance by compressing the neighborhood of each vertex using minhash signatures and facilitate rapid queries through Locality Sensitive Hashing. These techniques reduce query times from hours using industrial desktop machines operating on the full graph to milliseconds on standard laptops. Our method allows exploration of strongly associated regions (i.e., communities) of large graphs in real-time on a laptop. It has been deployed in software that is actively used by social network analysts and offers another channel for media owners to monetize their data, helping them to continue to provide

  19. Real time hybridization studies by resonant waveguide gratings using nanopattern imaging for Single Nucleotide Polymorphism detection

    KAUST Repository

    Bougot-Robin, Kristelle

    2013-12-20

    2D imaging of biochips is particularly interesting for multiplex biosensing. Resonant properties allow label-free detection using the change of refractive index at the chip surface. We demonstrate a new principle of Scanning Of Resonance on Chip by Imaging (SORCI) based on spatial profiles of nanopatterns of resonant waveguide gratings (RWGs) and its embodiment in a fluidic chip for real-time biological studies. This scheme allows multiplexing of the resonance itself by providing nanopattern sensing areas in a bioarray format. Through several chip designs we discuss resonance spatial profiles, dispersion and electric field distribution for optimal light-matter interaction with biological species of different sizes. Fluidic integration is carried out with a black anodized aluminum chamber, advantageous in term of mechanical stability, multiple uses of the chip, temperature control and low optical background. Real-time hybridization experiments are illustrated by SNP (Single Nucleotide Polymorphism) detection in gyrase A of E. coli K12, observed in evolution studies of resistance to the antibiotic ciprofloxacin. We choose a 100 base pairs (bp) DNA target (∼30 kDa) including the codon of interest and demonstrate the high specificity of our technique for probes and targets with close affinity constants. This work validates the safe applicability of our unique combination of RWGs and simple instrumentation for real-time biosensing with sensitivity in buffer solution of ∼10 pg/mm2. Paralleling the success of RWGs sensing for cells sensing, our work opens new avenues for a large number of biological studies. © 2013 Springer Science+Business Media.

  20. Detection of viable Salmonella in ice cream by TaqMan real-time polymerase chain reaction assay combining propidium monoazide

    Directory of Open Access Journals (Sweden)

    Yuexia Wang

    2015-09-01

    Full Text Available Real-time polymerase chain reaction (PCR allows rapid detection of Salmonella in frozen dairy products, but it might cause a false positive detection result because it might amplify DNA from dead target cells as well. In this study, Salmonella-free frozen ice cream was initially inoculated with heat-killed Salmonella Typhimurium cells and stored at −18°C. Bacterial DNA extracted from the sample was amplified using TaqMan probe-based real-time PCR targeting the invA gene. Our results indicated that DNA from the dead cells remained stable in frozen ice cream for at least 20 days, and could produce fluorescence signal for real-time PCR as well. To overcome this limitation, propidium monoazide (PMA was combined with real-time PCR. PMA treatment can effectively prevent PCR amplification from heat-killed Salmonella cells in frozen ice cream. The PMA real-time PCR assay can selectively detect viable Salmonella at as low as 103 CFU/mL. Combining 18 hours of pre-enrichment with the assay allows for the detection of viable Salmonella at 100 CFU/mL and avoiding the false-positive result of dead cells. The PMA real-time PCR assay provides an alternative specifically for detection of viable Salmonella in ice cream. However, when the PMA real-time PCR assay was evaluated in ice cream subjected to frozen storage, it obviously underestimated the contamination situation of viable Salmonella, which might lead to a false negative result. According to this result, the use of enrichment prior to PMA real-time PCR analysis remains as the more appropriate approach.

  1. Detection of viable Salmonella in ice cream by TaqMan real-time polymerase chain reaction assay combining propidium monoazide.

    Science.gov (United States)

    Wang, Yuexia; Yang, Ming; Liu, Shuchun; Chen, Wanyi; Suo, Biao

    2015-09-01

    Real-time polymerase chain reaction (PCR) allows rapid detection of Salmonella in frozen dairy products, but it might cause a false positive detection result because it might amplify DNA from dead target cells as well. In this study, Salmonella-free frozen ice cream was initially inoculated with heat-killed Salmonella Typhimurium cells and stored at -18°C. Bacterial DNA extracted from the sample was amplified using TaqMan probe-based real-time PCR targeting the invA gene. Our results indicated that DNA from the dead cells remained stable in frozen ice cream for at least 20 days, and could produce fluorescence signal for real-time PCR as well. To overcome this limitation, propidium monoazide (PMA) was combined with real-time PCR. PMA treatment can effectively prevent PCR amplification from heat-killed Salmonella cells in frozen ice cream. The PMA real-time PCR assay can selectively detect viable Salmonella at as low as 10 3  CFU/mL. Combining 18 hours of pre-enrichment with the assay allows for the detection of viable Salmonella at 10 0  CFU/mL and avoiding the false-positive result of dead cells. The PMA real-time PCR assay provides an alternative specifically for detection of viable Salmonella in ice cream. However, when the PMA real-time PCR assay was evaluated in ice cream subjected to frozen storage, it obviously underestimated the contamination situation of viable Salmonella, which might lead to a false negative result. According to this result, the use of enrichment prior to PMA real-time PCR analysis remains as the more appropriate approach. Copyright © 2015. Published by Elsevier B.V.

  2. Real time in situ detection of organic nitrates in atmospheric aerosols.

    Science.gov (United States)

    Rollins, Andrew W; Smith, Jared D; Wilson, Kevin R; Cohen, Ronald C

    2010-07-15

    A novel instrument is described that quantifies total particle-phase organic nitrates in real time with a detection limit of 0.11 microg m(-3) min(-1), 45 ppt min(-1) (-ONO(2)). Aerosol nitrates are separated from gas-phase nitrates with a short residence time activated carbon denuder. Detection of organic molecules containing -ONO(2) subunits is accomplished using thermal dissociation coupled to laser induced fluorescence detection of NO(2). This instrument is capable of high time resolution (seconds) measurements of particle-phase organic nitrates, without interference from inorganic nitrate. Here we use it to quantify organic nitrates in secondary organic aerosol generated from high-NO(x) photooxidation of limonene, alpha-pinene, Delta-3-carene, and tridecane. In these experiments the organic nitrate moiety is observed to be 6-15% of the total SOA mass.

  3. Investigation Model for DDoS Attack Detection in Real-Time

    Directory of Open Access Journals (Sweden)

    Abdulghani Ali Ahmed

    2015-02-01

    Full Text Available Investigating traffic of distributed denial of services (DDoS attack requires extra overhead which mostly results in network performance degradation. This study proposes an investigation model for detecting DDoS attack in real-time without causing negative degradation against network performance. The model investigates network traffic in a scalable way to detect user violations on quality of service regulations. Traffic investigation is triggered only when the network is congested; at that exact moment, burst gateways actually generate a congestion notification to misbehaving users. The misbehaving users are thus further investigated by measuring their consumption ratios of bandwidth. By exceeding the service level agreement bandwidth ratio, user traffic is filtered as DDoS traffic. Simulation results demonstrate that the proposed model efficiently monitors intrusive traffic and precisely detects DDoS attack.

  4. A method of real-time detection for distant moving obstacles by monocular vision

    Science.gov (United States)

    Jia, Bao-zhi; Zhu, Ming

    2013-12-01

    In this paper, we propose an approach for detection of distant moving obstacles like cars and bicycles by a monocular camera to cooperate with ultrasonic sensors in low-cost condition. We are aiming at detecting distant obstacles that move toward our autonomous navigation car in order to give alarm and keep away from them. Method of frame differencing is applied to find obstacles after compensation of camera's ego-motion. Meanwhile, each obstacle is separated from others in an independent area and given a confidence level to indicate whether it is coming closer. The results on an open dataset and our own autonomous navigation car have proved that the method is effective for detection of distant moving obstacles in real-time.

  5. Real-Time WGS-based Typing of VTEC Isolates for Surveillance and Outbreak Detection

    DEFF Research Database (Denmark)

    Joensen, Katrine Grimstrup; Hasman, Henrik; Scheutz, F.

    2013-01-01

    the IonTorrent PGM benchtop sequencing technology. WGS-based typing was carried out using web-based tools, developed by the Center for Genomic Epidemiology (www.genomicepidemiology.org), for determination of MLST types, virulence genes and phylogenetic relationship between the isolates. The WGS-based...... a small outbreak occurred. For all isolates, apart from one resulting in poor sequence output, the WGS-based typing led to detection of the same virulence gene variants as the routine typing, and was also able to detect many other possible virulence features, and in most instances produce a useful typing...... result faster than routine typing. Also, the WGS-approach was able to correctly detect, according to the routine typing, the isolates belonging to the outbreak. Conclusion: The real-time WGS-based typing was able to produce typing results comparable to the routine typing, at least as fast as the routine...

  6. Near-infrared high-resolution real-time omnidirectional imaging platform for drone detection

    Science.gov (United States)

    Popovic, Vladan; Ott, Beat; Wellig, Peter; Leblebici, Yusuf

    2016-10-01

    Recent technological advancements in hardware systems have made higher quality cameras. State of the art panoramic systems use them to produce videos with a resolution of 9000 x 2400 pixels at a rate of 30 frames per second (fps).1 Many modern applications use object tracking to determine the speed and the path taken by each object moving through a scene. The detection requires detailed pixel analysis between two frames. In fields like surveillance systems or crowd analysis, this must be achieved in real time.2 In this paper, we focus on the system-level design of multi-camera sensor acquiring near-infrared (NIR) spectrum and its ability to detect mini-UAVs in a representative rural Swiss environment. The presented results show the UAV detection from the trial that we conducted during a field trial in August 2015.

  7. The use of newly developed real-time PCR for the rapid identification of bacteria in culture-negative osteomyelitis.

    Science.gov (United States)

    Kobayashi, Naomi; Bauer, Thomas W; Sakai, Hiroshige; Togawa, Daisuke; Lieberman, Isador H; Fujishiro, Takaaki; Procop, Gary W

    2006-12-01

    We report a case of a culture-negative osteomyelitis in which our newly developed real-time polymerase chain reaction (PCR) could differentiate Staphylococcus aureus from Staphylococcus epidermidis. This is the first report that described the application of this novel assay to an orthopedics clinical sample. This assay may be useful for other clinical culture-negative cases in a combination with a broad-spectrum assay as a rapid microorganism identification method.

  8. Real-time portable system for fabric defect detection using an ARM processor

    Science.gov (United States)

    Fernandez-Gallego, J. A.; Yañez-Puentes, J. P.; Ortiz-Jaramillo, B.; Alvarez, J.; Orjuela-Vargas, S. A.; Philips, W.

    2012-06-01

    Modern textile industry seeks to produce textiles as little defective as possible since the presence of defects can decrease the final price of products from 45% to 65%. Automated visual inspection (AVI) systems, based on image analysis, have become an important alternative for replacing traditional inspections methods that involve human tasks. An AVI system gives the advantage of repeatability when implemented within defined constrains, offering more objective and reliable results for particular tasks than human inspection. Costs of automated inspection systems development can be reduced using modular solutions with embedded systems, in which an important advantage is the low energy consumption. Among the possibilities for developing embedded systems, the ARM processor has been explored for acquisition, monitoring and simple signal processing tasks. In a recent approach we have explored the use of the ARM processor for defects detection by implementing the wavelet transform. However, the computation speed of the preprocessing was not yet sufficient for real time applications. In this approach we significantly improve the preprocessing speed of the algorithm, by optimizing matrix operations, such that it is adequate for a real time application. The system was tested for defect detection using different defect types. The paper is focused in giving a detailed description of the basis of the algorithm implementation, such that other algorithms may use of the ARM operations for fast implementations.

  9. Effect of ionizing radiation on the quantitative detection of Salmonella using real-time PCR

    Energy Technology Data Exchange (ETDEWEB)

    Lim, Sangyong; Jung, Jinwoo [Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of); Kim, Minjeong; Ryu, Sangryeol [Department of Food and Animal Biotechnology, School of Agricultural Biotechnology, Center for Agricultural Biomaterials, Seoul National University, Seoul 151-921 (Korea, Republic of); Kim, Dongho [Radiation Research Center for Biotechnology, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of)], E-mail: fungikim@kaeri.re.kr

    2008-09-15

    Food irradiation is an economically viable technology for inactivating foodborne pathogens, but irradiation can mask pathogens in unhygienically prepared food. The aim of this study was to investigate the effect of irradiation treatment on the detection of Salmonella using real-time PCR. Three commercially available kits were tested, of which the InstaGene Matrix procedure was most effective in preparing template DNA from Salmonella exposed to radiation in broth culture. The minimum level of detection by real-time PCR combined with InstaGene Matrix was 3 log units of Salmonella per milliliter. However, when pure cultures of Salmonella were irradiated at 3 and 5 kGy, the cycle threshold (C{sub T}) increased 1-1.5-fold compared to irradiation at 0 and 1 kGy. This indicated that irradiation treatment may result in an underestimation of bacterial counts due to radiation-induced DNA lesions. We also compared C{sub T} values in inoculated chicken homogenates before and after irradiation, which in this model caused a 1.3-3.3-fold underestimation of bacterial counts with respect to irradiation dose.

  10. A generic method for real time detection of magnetic sensor failure on tokamaks

    International Nuclear Information System (INIS)

    Nouailletas, Rémy; Moreau, Philippe; Bremond, Sylvain

    2012-01-01

    Highlights: ► We propose a generic method to detect and correct in real time faults on magnetic sensor. ► This method is applied to Tore Supra and tested offline with real data. ► Then the method is modified to be applied to ITER ex-vessel sensor configuration. ► The method is tested on the ITER case with simulated data. - Abstract: In tokamaks, magnetic field probe sensors are used to measure the plasma position. If a sensor provides a wrong data, the error may propagate through the control loop and cause undesirable contact between the vessel wall and the plasma. In the case of a tokamak with water cooled walls, these types of event may be very serious. Despite of these unlikely faults, the potential damages call for a real time check of magnetic sensor data before using them for control. In this paper a simple and generic method based on the comparison of each sensor to a weighted sum of its neighbors is proposed. From the analysis of the residue (the result of the comparison), the fault can be detected and compensated. The method is tuned and tested against Tore Supra experimental data. Then, the method is adapted to ITER and assessed on a reference ITER scenario using simulated magnetic sensor data.

  11. Real-time QRS detection using integrated variance for ECG gated cardiac MRI

    Directory of Open Access Journals (Sweden)

    Schmidt Marcus

    2016-09-01

    Full Text Available During magnetic resonance imaging (MRI, a patient’s vital signs are required for different purposes. In cardiac MRI (CMR, an electrocardiogram (ECG of the patient is required for triggering the image acquisition process. However, a reliable QRS detection of an ECG signal acquired inside an MRI scanner is a challenging task due to the magnetohydrodynamic (MHD effect which interferes with the ECG. The aim of this work was to develop a reliable QRS detector usable inside the MRI which also fulfills the standards for medical devices (IEC 60601-2-27. Therefore, a novel real-time QRS detector based on integrated variance measurements is presented. The algorithm was trained on ANSI/AAMI EC13 test waveforms and was then applied to two databases with 12-lead ECG signals recorded inside and outside an MRI scanner. Reliable results for both databases were achieved for the ECG signals recorded inside (DBMRI: sensitivity Se = 99.94%, positive predictive value +P = 99.84% and outside (DBInCarT: Se = 99.29%, +P = 99.72% the MRI. Due to the accurate R-peak detection in real-time this can be used for monitoring and triggering in MRI exams.

  12. An Algorithm for Real-Time Pulse Waveform Segmentation and Artifact Detection in Photoplethysmograms.

    Science.gov (United States)

    Fischer, Christoph; Domer, Benno; Wibmer, Thomas; Penzel, Thomas

    2017-03-01

    Photoplethysmography has been used in a wide range of medical devices for measuring oxygen saturation, cardiac output, assessing autonomic function, and detecting peripheral vascular disease. Artifacts can render the photoplethysmogram (PPG) useless. Thus, algorithms capable of identifying artifacts are critically important. However, the published PPG algorithms are limited in algorithm and study design. Therefore, the authors developed a novel embedded algorithm for real-time pulse waveform (PWF) segmentation and artifact detection based on a contour analysis in the time domain. This paper provides an overview about PWF and artifact classifications, presents the developed PWF analysis, and demonstrates the implementation on a 32-bit ARM core microcontroller. The PWF analysis was validated with data records from 63 subjects acquired in a sleep laboratory, ergometry laboratory, and intensive care unit in equal parts. The output of the algorithm was compared with harmonized experts' annotations of the PPG with a total duration of 31.5 h. The algorithm achieved a beat-to-beat comparison sensitivity of 99.6%, specificity of 90.5%, precision of 98.5%, and accuracy of 98.3%. The interrater agreement expressed as Cohen's kappa coefficient was 0.927 and as F-measure was 0.990. In conclusion, the PWF analysis seems to be a suitable method for PPG signal quality determination, real-time annotation, data compression, and calculation of additional pulse wave metrics such as amplitude, duration, and rise time.

  13. Real Time Polymerase Chain Reaction : Perangkat Diagnostic Alternatif untuk Melacak Virus Nipah (REAL TIME POLYMERASE CHAIN REACTION : AN ALTERNATIVE DIAGNOSTIC TOOL TO DETECT NIPAH VIRUS

    Directory of Open Access Journals (Sweden)

    Indrawati Sendow

    2014-05-01

    Full Text Available Nipah is a dangerous zoonotic disease with a high social, economical and psychological impact. Fruitbat Pteropus sp. is one of the nipah virus  reservoir host. As the virus is categorized as a dangerous zoonoticdisease that cause fatal in human, all works related to live virus should be conducted in a laboratory withBSL4 facilities. The detection of nipah virus using real time PCR to replace virus isolastion can thereforebe conducted in a laboratory without BSL4 facilities. The results was further  confirmed at referencelaboratory at   Australian Animal Health Laboratory ( AAHL Geelong, Australia, indicated that nipahvirus can be detected in saliva of fruit bat P. vampyrus in Medan North Sumatera.

  14. Detection of Schistosoma mansoni and Schistosoma haematobium by Real-Time PCR with High Resolution Melting Analysis

    Directory of Open Access Journals (Sweden)

    Hany Sady

    2015-07-01

    Full Text Available The present study describes a real-time PCR approach with high resolution melting-curve (HRM assay developed for the detection and differentiation of Schistosoma mansoni and S. haematobium in fecal and urine samples collected from rural Yemen. The samples were screened by microscopy and PCR for the Schistosoma species infection. A pair of degenerate primers were designed targeting partial regions in the cytochrome oxidase subunit I (cox1 gene of S. mansoni and S. haematobium using real-time PCR-HRM assay. The overall prevalence of schistosomiasis was 31.8%; 23.8% of the participants were infected with S. haematobium and 9.3% were infected with S. mansoni. With regards to the intensity of infections, 22.1% and 77.9% of S. haematobium infections were of heavy and light intensities, respectively. Likewise, 8.1%, 40.5% and 51.4% of S. mansoni infections were of heavy, moderate and light intensities, respectively. The melting points were distinctive for S. mansoni and S. haematobium, categorized by peaks of 76.49 ± 0.25 °C and 75.43 ± 0.26 °C, respectively. HRM analysis showed high detection capability through the amplification of Schistosoma DNA with as low as 0.0001 ng/µL. Significant negative correlations were reported between the real-time PCR-HRM cycle threshold (Ct values and microscopic egg counts for both S. mansoni in stool and S. haematobium in urine (p < 0.01. In conclusion, this closed-tube HRM protocol provides a potentially powerful screening molecular tool for the detection of S. mansoni and S. haematobium. It is a simple, rapid, accurate, and cost-effective method. Hence, this method is a good alternative approach to probe-based PCR assays.

  15. Real-time control of focused ultrasound heating based on rapid MR thermometry.

    Science.gov (United States)

    Vimeux, F C; De Zwart, J A; Palussiére, J; Fawaz, R; Delalande, C; Canioni, P; Grenier, N; Moonen, C T

    1999-03-01

    Real-time control of the heating procedure is essential for hyperthermia applications of focused ultrasound (FUS). The objective of this study is to demonstrate the feasibility of MRI-controlled FUS. An automatic control system was developed using a dedicated interface between the MR system control computer and the FUS wave generator. Two algorithms were used to regulate FUS power to maintain the focal point temperature at a desired level. Automatic control of FUS power level was demonstrated ex vivo at three target temperature levels (increase of 5 degrees C, 10 degrees C, and 30 degrees C above room temperature) during 30-minute hyperthermic periods. Preliminary in vivo results on rat leg muscle confirm that necrosis estimate, calculated on-line during FUS sonication, allows prediction of tissue damage. CONCLUSIONS. The feasibility of fully automatic FUS control based on MRI thermometry has been demonstrated.

  16. Real-time Quaking-induced Conversion Assay for Detection of CWD Prions in Fecal Material.

    Science.gov (United States)

    Cheng, Yo Ching; Hannaoui, Samia; John, Theodore Ralph; Dudas, Sandor; Czub, Stefanie; Gilch, Sabine

    2017-09-29

    The RT-QuIC technique is a sensitive in vitro cell-free prion amplification assay based mainly on the seeded misfolding and aggregation of recombinant prion protein (PrP) substrate using prion seeds as a template for the conversion. RT-QuIC is a novel high-throughput technique which is analogous to real-time polymerase chain reaction (PCR). Detection of amyloid fibril growth is based on the dye Thioflavin T, which fluoresces upon specific interaction with ᵦ-sheet rich proteins. Thus, amyloid formation can be detected in real time. We attempted to develop a reliable non-invasive screening test to detect chronic wasting disease (CWD) prions in fecal extract. Here, we have specifically adapted the RT-QuIC technique to reveal PrP Sc seeding activity in feces of CWD infected cervids. Initially, the seeding activity of the fecal extracts we prepared was relatively low in RT-QuIC, possibly due to potential assay inhibitors in the fecal material. To improve seeding activity of feces extracts and remove potential assay inhibitors, we homogenized the fecal samples in a buffer containing detergents and protease inhibitors. We also submitted the samples to different methodologies to concentrate PrP Sc on the basis of protein precipitation using sodium phosphotungstic acid, and centrifugal force. Finally, the feces extracts were tested by optimized RT-QuIC which included substrate replacement in the protocol to improve the sensitivity of detection. Thus, we established a protocol for sensitive detection of CWD prion seeding activity in feces of pre-clinical and clinical cervids by RT-QuIC, which can be a practical tool for non-invasive CWD diagnosis.

  17. Clinical Application of a Multiplex Real-Time PCR Assay for Simultaneous Detection of Legionella Species, Legionella pneumophila, and Legionella pneumophila Serogroup 1

    OpenAIRE

    Benitez, Alvaro J.; Winchell, Jonas M.

    2014-01-01

    We developed a single-tube multiplex real-time PCR assay capable of simultaneously detecting and discriminating Legionella spp., Legionella pneumophila, and Legionella pneumophila serogroup 1 in primary specimens. Evaluation of 21 clinical specimens and 115 clinical isolates demonstrated this assay to be a rapid, high-throughput diagnostic test with 100% specificity that may aid during legionellosis outbreaks and epidemiologic investigations.

  18. Development of a novel quantitative real-time RT-PCR assay for the simultaneous detection of all serotypes of Foot-and-mouth disease virus

    DEFF Research Database (Denmark)

    Rasmussen, Thomas Bruun; Uttenthal, Åse; de Stricker, K.

    2003-01-01

    Foot-and-mouth disease virus (FMDV) spreads extremely fast and the need for rapid and robust diagnostic virus detection systems was obvious during the recent European epidemic. Using a novel real-time RT-PCR system based on primer-probe energy transfer (PriProET) we present here an assay targeting...

  19. Real Time Corner Detection for Miniaturized Electro-Optical Sensors Onboard Small Unmanned Aerial Systems

    Directory of Open Access Journals (Sweden)

    Antonio Moccia

    2012-01-01

    Full Text Available This paper describes the target detection algorithm for the image processor of a vision-based system that is installed onboard an unmanned helicopter. It has been developed in the framework of a project of the French national aerospace research center Office National d’Etudes et de Recherches Aérospatiales (ONERA which aims at developing an air-to-ground target tracking mission in an unknown urban environment. In particular, the image processor must detect targets and estimate ground motion in proximity of the detected target position. Concerning the target detection function, the analysis has dealt with realizing a corner detection algorithm and selecting the best choices in terms of edge detection methods, filtering size and type and the more suitable criterion of detection of the points of interest in order to obtain a very fast algorithm which fulfills the computation load requirements. The compared criteria are the Harris-Stephen and the Shi-Tomasi, ones, which are the most widely used in literature among those based on intensity. Experimental results which illustrate the performance of the developed algorithm and demonstrate that the detection time is fully compliant with the requirements of the real-time system are discussed.

  20. Real-Time Ventricular Fibrillation Detection Using an Embedded Microcontroller in a Pervasive Environment

    Directory of Open Access Journals (Sweden)

    Sundeok Kwon

    2018-06-01

    Full Text Available Many healthcare problems are life threatening and need real-time detection to improve patient safety. Heart attack or ventricular fibrillation (VF is a common problem worldwide. Most previous research on VF detection has used ECG devices to capture data and sent to other higher performance units for processing and has relied on domain experts and/or sophisticated algorithms for detection. In this case, it delayed the response time and consumed much more energy of the ECG module. In this study, we propose a prototype that an embedded microcontroller where an ECG sensor is used to capture, filter and process data, run VF detection algorithms, and only transmit the detected event to the smartphone for alert and call for services. We discuss how to adapt a common filtering and scale process and five light-weighted algorithms from open literature to realize the idea. We also develop an integrated prototype, which emulates the VF process from existing data sets, to evaluate the detection capability of the framework and algorithms. Our results show that (1 TD outperforms the other four algorithms considered with sensitivity reaching 96.56% and specificity reaching 81.53% in the MIT-BIH dataset. Our evaluations confirm that with some adaptation the conventional filtering process and detection algorithms can be efficiently deployed in a microcontroller with good detection accuracy while saving battery power, shortening response time, and conserving the network bandwidth.

  1. EVA GREEN REAL-TIME PCR USED TO DETECT CELERY AS AN ALLERGEN IN FOOD

    Directory of Open Access Journals (Sweden)

    Ondrej Škultéty

    2011-04-01

    Full Text Available EvaGreen®  Real-Time PCR method has been used for celery(Apium graveolens allergen detection. A primer designed in mannitol dehydrogenase gene region has been used for specific celery identification in sample. The results show possibility to create calibration curve using artificially adulterated samples. The increasing variability between parallel calibration of celery samples has been observed from 0.1 % to 100%. Detection limit has been set to value 0.1% in celery representing 1000 ppm. Fluorescent signal has been presented even in samples with lower percentage addition of celery but these samples have been excluded according to unspecific melting curve.doi:10.5219/138

  2. Real-Time Detection of Rupture Development: Earthquake Early Warning Using P Waves From Growing Ruptures

    Science.gov (United States)

    Kodera, Yuki

    2018-01-01

    Large earthquakes with long rupture durations emit P wave energy throughout the rupture period. Incorporating late-onset P waves into earthquake early warning (EEW) algorithms could contribute to robust predictions of strong ground motion. Here I describe a technique to detect in real time P waves from growing ruptures to improve the timeliness of an EEW algorithm based on seismic wavefield estimation. The proposed P wave detector, which employs a simple polarization analysis, successfully detected P waves from strong motion generation areas of the 2011 Mw 9.0 Tohoku-oki earthquake rupture. An analysis using 23 large (M ≥ 7) events from Japan confirmed that seismic intensity predictions based on the P wave detector significantly increased lead times without appreciably decreasing the prediction accuracy. P waves from growing ruptures, being one of the fastest carriers of information on ongoing rupture development, have the potential to improve the performance of EEW systems.

  3. A novel time-domain signal processing algorithm for real time ventricular fibrillation detection

    International Nuclear Information System (INIS)

    Monte, G E; Scarone, N C; Liscovsky, P O; Rotter, P

    2011-01-01

    This paper presents an application of a novel algorithm for real time detection of ECG pathologies, especially ventricular fibrillation. It is based on segmentation and labeling process of an oversampled signal. After this treatment, analyzing sequence of segments, global signal behaviours are obtained in the same way like a human being does. The entire process can be seen as a morphological filtering after a smart data sampling. The algorithm does not require any ECG digital signal pre-processing, and the computational cost is low, so it can be embedded into the sensors for wearable and permanent applications. The proposed algorithms could be the input signal description to expert systems or to artificial intelligence software in order to detect other pathologies.

  4. A novel time-domain signal processing algorithm for real time ventricular fibrillation detection

    Science.gov (United States)

    Monte, G. E.; Scarone, N. C.; Liscovsky, P. O.; Rotter S/N, P.

    2011-12-01

    This paper presents an application of a novel algorithm for real time detection of ECG pathologies, especially ventricular fibrillation. It is based on segmentation and labeling process of an oversampled signal. After this treatment, analyzing sequence of segments, global signal behaviours are obtained in the same way like a human being does. The entire process can be seen as a morphological filtering after a smart data sampling. The algorithm does not require any ECG digital signal pre-processing, and the computational cost is low, so it can be embedded into the sensors for wearable and permanent applications. The proposed algorithms could be the input signal description to expert systems or to artificial intelligence software in order to detect other pathologies.

  5. Real-Time PCR Detection of Dogwood Anthracnose Fungus in Historical Herbarium Specimens from Asia.

    Science.gov (United States)

    Miller, Stephen; Masuya, Hayato; Zhang, Jian; Walsh, Emily; Zhang, Ning

    2016-01-01

    Cornus species (dogwoods) are popular ornamental trees and important understory plants in natural forests of northern hemisphere. Dogwood anthracnose, one of the major diseases affecting the native North American Cornus species, such as C. florida, is caused by the fungal pathogen Discula destructiva. The origin of this fungus is not known, but it is hypothesized that it was imported to North America with its host plants from Asia. In this study, a TaqMan real-time PCR assay was used to detect D. destructiva in dried herbarium and fresh Cornus samples. Several herbarium specimens from Japan and China were detected positive for D. destructiva, some of which were collected before the first report of the dogwood anthracnose in North America. Our findings further support that D. destructiva was introduced to North America from Asia where the fungus likely does not cause severe disease.

  6. Development of real-time PCR for detection of Mycoplasma hominis

    DEFF Research Database (Denmark)

    Baczynska, A.; Svenstrup, Helle Friis; Fedder, J.

    2004-01-01

    BACKGROUND: Mycoplasma hominis is associated with pelvic inflammatory disease, bacterial vaginosis, post partum fever, sepsis and infections of the central nervous system often leading to serious conditions. Association with development of female infertility has also been suggested, but different....... Information on bacterial load in genital swabs can be obtained. The assay allowed detection of M. hominis in a closed system reducing the risk of contamination by amplicon carry-over......., glyceraldehyde-3-phosphate dehydrogenase (gap), as a target. RESULTS: Real-time PCR was optimized to detect 10 copies of M. hominis PG21 genomic DNA. A fluorescence signal was measured for all 20 other M. hominis isolates, and melting curves analysis showed variations in the melting temperature in agreement...

  7. 3D Printed "Earable" Smart Devices for Real-Time Detection of Core Body Temperature.

    Science.gov (United States)

    Ota, Hiroki; Chao, Minghan; Gao, Yuji; Wu, Eric; Tai, Li-Chia; Chen, Kevin; Matsuoka, Yasutomo; Iwai, Kosuke; Fahad, Hossain M; Gao, Wei; Nyein, Hnin Yin Yin; Lin, Liwei; Javey, Ali

    2017-07-28

    Real-time detection of basic physiological parameters such as blood pressure and heart rate is an important target in wearable smart devices for healthcare. Among these, the core body temperature is one of the most important basic medical indicators of fever, insomnia, fatigue, metabolic functionality, and depression. However, traditional wearable temperature sensors are based upon the measurement of skin temperature, which can vary dramatically from the true core body temperature. Here, we demonstrate a three-dimensional (3D) printed wearable "earable" smart device that is designed to be worn on the ear to track core body temperature from the tympanic membrane (i.e., ear drum) based on an infrared sensor. The device is fully integrated with data processing circuits and a wireless module for standalone functionality. Using this smart earable device, we demonstrate that the core body temperature can be accurately monitored regardless of the environment and activity of the user. In addition, a microphone and actuator are also integrated so that the device can also function as a bone conduction hearing aid. Using 3D printing as the fabrication method enables the device to be customized for the wearer for more personalized healthcare. This smart device provides an important advance in realizing personalized health care by enabling real-time monitoring of one of the most important medical parameters, core body temperature, employed in preliminary medical screening tests.

  8. Real-time detection of natural objects using AM-coded spectral matching imager

    Science.gov (United States)

    Kimachi, Akira

    2005-01-01

    This paper describes application of the amplitude-modulation (AM)-coded spectral matching imager (SMI) to real-time detection of natural objects such as human beings, animals, vegetables, or geological objects or phenomena, which are much more liable to change with time than artificial products while often exhibiting characteristic spectral functions associated with some specific activity states. The AM-SMI produces correlation between spectral functions of the object and a reference at each pixel of the correlation image sensor (CIS) in every frame, based on orthogonal amplitude modulation (AM) of each spectral channel and simultaneous demodulation of all channels on the CIS. This principle makes the SMI suitable to monitoring dynamic behavior of natural objects in real-time by looking at a particular spectral reflectance or transmittance function. A twelve-channel multispectral light source was developed with improved spatial uniformity of spectral irradiance compared to a previous one. Experimental results of spectral matching imaging of human skin and vegetable leaves are demonstrated, as well as a preliminary feasibility test of imaging a reflective object using a test color chart.

  9. The use of bivalves as rapid, real-time indicators of aquatic pollution

    International Nuclear Information System (INIS)

    Markich, S.J.

    1995-01-01

    The ability of bivalves to filter large volumes of water on a daily basis, combined with the relatively high permeability of their cell membranes, make them valuable organisms to use in the contemporary detection of pollution. Bivalves are well known to respond to chemical contaminants by isolating their soft tissues from the aquatic medium by valve closure. The sensory acuity (via specialized sensory regions including the osphradium) and associated repertoire of this behavioral response can be employed to assess subtle effects exerted by chemical contaminants, such as complex effluents, that may ultimately influence the survival of these organisms. As hazard assessment tools, behavioral studies reflect sublethal toxicity and often yield a highly sensitive estimate of the lowest observable effect concentration (LOEC). Moreover, valve movement behavior has been identified as one of the more sensitive biological early warning measures to a variety of aquatic contaminants, in comparison with those used in other aquatic animal phyla. Therefore, the valve movement behavior of both freshwater (Hyridella depressa, Velesunio angasi and V. ambiguus) and marine (Mytilus edulis) bivalves was continuously monitored, using an on-line computer based data acquisition system, during exposure to either trace metals (e.g. Cu, Cd, Mn and U) or complex effluents (ie treated sewage effluent and acid leachate derived from contaminated Sydney Harbour sediments), in the context of using the valve movement behavior of the bivalve species to indicate the biological significance of exposure to the above-mentioned pollutants. The results indicate that several components of the valve movement behavior of each bivalve provide quantifiable and ecologically interpretable sub-lethal endpoints for the rapid and sensitive evaluation of waters containing either complex effluents or elevated levels of trace metals

  10. Optics based signal processing methods for intraoperative blood vessel detection and quantification in real time (Conference Presentation)

    Science.gov (United States)

    Chaturvedi, Amal; Shukair, Shetha A.; Le Rolland, Paul; Vijayvergia, Mayank; Subramanian, Hariharan; Gunn, Jonathan W.

    2016-03-01

    Minimally invasive operations require surgeons to make difficult cuts to blood vessels and other tissues with impaired tactile and visual feedback. This leads to inadvertent cuts to blood vessels hidden beneath tissue, causing serious health risks to patients and a non-reimbursable financial burden to hospitals. Intraoperative imaging technologies have been developed, but these expensive systems can be cumbersome and provide only a high-level view of blood vessel networks. In this research, we propose a lean reflectance-based system, comprised of a dual wavelength LED, photodiode, and novel signal processing algorithms for rapid vessel characterization. Since this system takes advantage of the inherent pulsatile light absorption characteristics of blood vessels, no contrast agent is required for its ability to detect the presence of a blood vessel buried deep inside any tissue type (up to a cm) in real time. Once a vessel is detected, the system is able to estimate the distance of the vessel from the probe and the diameter size of the vessel (with a resolution of ~2mm), as well as delineate the type of tissue surrounding the vessel. The system is low-cost, functions in real-time, and could be mounted on already existing surgical tools, such as Kittner dissectors or laparoscopic suction irrigation cannulae. Having been successfully validated ex vivo, this technology will next be tested in a live porcine study and eventually in clinical trials.

  11. GMO detection using a bioluminescent real time reporter (BART of loop mediated isothermal amplification (LAMP suitable for field use

    Directory of Open Access Journals (Sweden)

    Kiddle Guy

    2012-04-01

    Full Text Available Abstract Background There is an increasing need for quantitative technologies suitable for molecular detection in a variety of settings for applications including food traceability and monitoring of genetically modified (GM crops and their products through the food processing chain. Conventional molecular diagnostics utilising real-time polymerase chain reaction (RT-PCR and fluorescence-based determination of amplification require temperature cycling and relatively complex optics. In contrast, isothermal amplification coupled to a bioluminescent output produced in real-time (BART occurs at a constant temperature and only requires a simple light detection and integration device. Results Loop mediated isothermal amplification (LAMP shows robustness to sample-derived inhibitors. Here we show the applicability of coupled LAMP and BART reactions (LAMP-BART for determination of genetically modified (GM maize target DNA at low levels of contamination (0.1-5.0% GM using certified reference material, and compare this to RT-PCR. Results show that conventional DNA extraction methods developed for PCR may not be optimal for LAMP-BART quantification. Additionally, we demonstrate that LAMP is more tolerant to plant sample-derived inhibitors, and show this can be exploited to develop rapid extraction techniques suitable for simple field-based qualitative tests for GM status determination. We also assess the effect of total DNA assay load on LAMP-BART quantitation. Conclusions LAMP-BART is an effective and sensitive technique for GM detection with significant potential for quantification even at low levels of contamination and in samples derived from crops such as maize with a large genome size. The resilience of LAMP-BART to acidic polysaccharides makes it well suited to rapid sample preparation techniques and hence to both high throughput laboratory settings and to portable GM detection applications. The impact of the plant sample matrix and genome loading

  12. Network-Based Real-time Integrated Fire Detection and Alarm (FDA) System with Building Automation

    Science.gov (United States)

    Anwar, F.; Boby, R. I.; Rashid, M. M.; Alam, M. M.; Shaikh, Z.

    2017-11-01

    Fire alarm systems have become increasingly an important lifesaving technology in many aspects, such as applications to detect, monitor and control any fire hazard. A large sum of money is being spent annually to install and maintain the fire alarm systems in buildings to protect property and lives from the unexpected spread of fire. Several methods are already developed and it is improving on a daily basis to reduce the cost as well as increase quality. An integrated Fire Detection and Alarm (FDA) systems with building automation was studied, to reduce cost and improve their reliability by preventing false alarm. This work proposes an improved framework for FDA system to ensure a robust intelligent network of FDA control panels in real-time. A shortest path algorithmic was chosen for series of buildings connected by fiber optic network. The framework shares information and communicates with each fire alarm panels connected in peer to peer configuration and declare the network state using network address declaration from any building connected in network. The fiber-optic connection was proposed to reduce signal noises, thus increasing large area coverage, real-time communication and long-term safety. Based on this proposed method an experimental setup was designed and a prototype system was developed to validate the performance in practice. Also, the distributed network system was proposed to connect with an optional remote monitoring terminal panel to validate proposed network performance and ensure fire survivability where the information is sequentially transmitted. The proposed FDA system is different from traditional fire alarm and detection system in terms of topology as it manages group of buildings in an optimal and efficient manner.Introduction

  13. The Prototype of Real-time Object Detection System Based on SMS

    Directory of Open Access Journals (Sweden)

    M. Hana Mirza

    2010-08-01

    Full Text Available The powerful algorithm to detect object movement in development of room monitoring system is very urgent. The commond algorithm needs complex computation. In this research, the prototype of real-time object detection system using simple algorithm is developed, i.e. using the determination of the max noise/pixel value and the tolerance threshold of image accurately, and then the system automatically send a SMS (short message services to user when the object movement is detected. The developed prototype used a Logitech QuickCam webcam, a Siemens C45 mobile phone and a data cable, and the Borland Delphi 7 with additional components and Serial PortNG Tvideo as system software. The application also includes a database to store the captured images whenever object movement is detected. The test results by varying conditions of light intensities using a 5-watt light bulb, fluorescent lamp 20 and 40 watts indicate that the application is able to automatically detect the presence of moving objects with 100% success rate. The success rate is strongly influenced by the determination of the max noise/pixel value and the tolerance threshold during system configuration. This application is also capable of sending SMS automatically when the system detects a moving object with an average time of 8.35 seconds.

  14. Real Time Supervisors and Monitors for Performing Health Monitoring and Fault Detection for Systems Operating in Multiple Regimes

    National Research Council Canada - National Science Library

    Jaw, Link

    2003-01-01

    In this Phase I STTR, SMI and ARL have developed a Real Time Supervisor for fault detection and system reconfiguration in a team of micro UAVs, that are tasked to perform a team mission like surveillance or rendezvous...

  15. Detection of Mycoplasma genitalium in female cervical samples by Multitarget Real-Time PCR

    Directory of Open Access Journals (Sweden)

    Sabina Mahmutović-Vranić

    2007-05-01

    Full Text Available Mycoplasma genitalum (MG is associated with variety of urogenital infections such as non-gonococcal urethritis (NGU, endometritis and cervicitis. The objective of this study was to demonstrate and evaluate a research polymerase chain reaction (PCR assay, for the detection of MG in cervical samples of a tested population of women attending gynecology clinics in Bosnia and Herzegovina. The Multitarget Real-Time (MTRT PCR, utilizing the ABI 7900HT, the sequence detection system, was performed for the detection of MG. Cervical samples (N=97 from females were divided into three types of patient groups: Group 1: patients who had known abnormal clinical cytology reports (N=34; Group 2: patients who reported a history of genitourinary infections (N=22; and Group 3: patients not in either groups 1 or 2 (N=41. Overall, 14,43% (14/97 of those tested were positive for MG. A positive sample was defined as having a cycle threshold cross point (Ct < 40,0 with a fluorescent detection comparable to the low positive control utilized during the run. This study validated the use of MTRT PCR as a reliable method for the detection of MG in clinical specimens and should facilitate large-scale screening for this organism.

  16. Ring trial 2016 for Bluetongue virus detection by real-time RT-PCR in France.

    Science.gov (United States)

    Sailleau, Corinne; Viarouge, Cyril; Breard, Emmanuel; Vitour, Damien; Zientara, Stephan

    2017-05-01

    Since the unexpected emergence of BTV-8 in Northern Europe and the incursion of BTV-8 and 1 in France in 2006-2007, molecular diagnosis has considerably evolved. Several real-time RT-PCR (rtRT-PCR) methods have been developed and published, and are currently being used in many countries across Europe for BTV detection and typing. In France, the national reference laboratory (NRL) for orbiviruses develops and validates 'ready-to-use' kits with private companies for viral RNA detection. The regional laboratories network that was set up to deal with a heavy demand for analyses has used these available kits. From 2007, ring tests were organized to monitor the performance of the French laboratories. This study presents the results of 63 regional laboratories in the ring trial organized in 2016. Blood samples were sent to the laboratories. Participants were asked to use the rtRT-PCR methods in place in their laboratory, for detection of all BTV serotypes and specifically BTV-8. The French regional laboratories are able to detect and genotype BTV in affected animals. Despite the use of several methods (i.e. RNA extraction and different commercial rtRT-PCRs), the network is homogeneous. The ring trial demonstrated that the French regional veterinary laboratories have reliable and robust BTV diagnostic tools for BTV genome detection.

  17. A method for real-time memory efficient implementation of blob detection in large images

    Directory of Open Access Journals (Sweden)

    Petrović Vladimir L.

    2017-01-01

    Full Text Available In this paper we propose a method for real-time blob detection in large images with low memory cost. The method is suitable for implementation on the specialized parallel hardware such as multi-core platforms, FPGA and ASIC. It uses parallelism to speed-up the blob detection. The input image is divided into blocks of equal sizes to which the maximally stable extremal regions (MSER blob detector is applied in parallel. We propose the usage of multiresolution analysis for detection of large blobs which are not detected by processing the small blocks. This method can find its place in many applications such as medical imaging, text recognition, as well as video surveillance or wide area motion imagery (WAMI. We explored the possibilities of usage of detected blobs in the feature-based image alignment as well. When large images are processed, our approach is 10 to over 20 times more memory efficient than the state of the art hardware implementation of the MSER.

  18. Ionoluminescence analysis of glass scintillators and application to single-ion-hit real-time detection

    Energy Technology Data Exchange (ETDEWEB)

    Yokoyama, Akihito, E-mail: yokoyama.akihito@jaea.go.jp [Graduate School of Science and Technology, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515 (Japan); Takasaki Advanced Radiation Research Institute (TARRI), Japan Atomic Energy Agency (JAEA), 1233 Watanuki-machi, Takasaki, Gunma 370-1292 (Japan); Kada, Wataru [Graduate School of Science and Technology, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515 (Japan); Satoh, Takahiro; Koka, Masashi [Takasaki Advanced Radiation Research Institute (TARRI), Japan Atomic Energy Agency (JAEA), 1233 Watanuki-machi, Takasaki, Gunma 370-1292 (Japan); Shimada, Keisuke; Yokoata, Yuya; Miura, Kenta; Hanaizumi, Osamu [Graduate School of Science and Technology, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515 (Japan)

    2016-03-15

    In this paper, we propose and test a real-time detection system for single-ion hits using mega-electronvolt (MeV)-heavy ions. The system was constructed using G2000 and G9 glass scintillators, as well as an electron-multiplying charge-coupled device (EMCCD) camera combined with an inverted microscope with a 10× objective lens. Commercially available G2000 and G9 glass scintillators, which have been reported to exhibit strong photoluminescence at 489, 543, 585, and 622 nm as a result of the Tb{sup 3+} f–f transition, were employed for highly accurate ionized particle detection. The EMCCD camera had a resolution of 512 × 512 pixels, each with a size of 16 μm × 16 μm, and a maximum linear gain of 8 × 10{sup 5} electrons. For 260-MeV Ne, 3 ion hits/s were detected by our system. The intensity of the ionoluminescence (IL) peak induced by the heavy ions was 140 times the noise intensity. In contrast, the luminous diameter at the full width at half maximum (FWHM) in both the horizontal and vertical directions was calculated to be approximately 4.5 μm. These results suggest that our detection system can accurately detect single-ion hits with a diameter of the order of 1 μm.

  19. Implementation of a General Real-Time Visual Anomaly Detection System Via Soft Computing

    Science.gov (United States)

    Dominguez, Jesus A.; Klinko, Steve; Ferrell, Bob; Steinrock, Todd (Technical Monitor)

    2001-01-01

    The intelligent visual system detects anomalies or defects in real time under normal lighting operating conditions. The application is basically a learning machine that integrates fuzzy logic (FL), artificial neural network (ANN), and generic algorithm (GA) schemes to process the image, run the learning process, and finally detect the anomalies or defects. The system acquires the image, performs segmentation to separate the object being tested from the background, preprocesses the image using fuzzy reasoning, performs the final segmentation using fuzzy reasoning techniques to retrieve regions with potential anomalies or defects, and finally retrieves them using a learning model built via ANN and GA techniques. FL provides a powerful framework for knowledge representation and overcomes uncertainty and vagueness typically found in image analysis. ANN provides learning capabilities, and GA leads to robust learning results. An application prototype currently runs on a regular PC under Windows NT, and preliminary work has been performed to build an embedded version with multiple image processors. The application prototype is being tested at the Kennedy Space Center (KSC), Florida, to visually detect anomalies along slide basket cables utilized by the astronauts to evacuate the NASA Shuttle launch pad in an emergency. The potential applications of this anomaly detection system in an open environment are quite wide. Another current, potentially viable application at NASA is in detecting anomalies of the NASA Space Shuttle Orbiter's radiator panels.

  20. First Real-Time Detection of Surface Dust in a Tokamak

    International Nuclear Information System (INIS)

    Skinner, C.; Rais, B.; Roquemore, A.L.; Kugel, H.W.; Marsala, R.; Provost, T.

    2010-01-01

    The first real-time detection of surface dust inside a tokamak was made using an electrostatic dust detector. A fine grid of interlocking circuit traces was installed in the NSTX vessel and biased to 50 v. Impinging dust particles created a temporary short circuit and the resulting current pulse was recorded by counting electronics. The techniques used to increase the detector sensitivity by a factor of x10,000 to match NSTX dust levels while suppressing electrical pickup are presented. The results were validated by comparison to lab measurements, by the null signal from a covered detector that was only sensitive to pickup, and by the dramatic increase in signal when Li particles were introduced for wall conditioning purposes.

  1. FRB microstructure revealed by the real-time detection of FRB170827

    Science.gov (United States)

    Farah, W.; Flynn, C.; Bailes, M.; Jameson, A.; Bannister, K. W.; Barr, E. D.; Bateman, T.; Bhandari, S.; Caleb, M.; Campbell-Wilson, D.; Chang, S.-W.; Deller, A.; Green, A. J.; Hunstead, R.; Jankowski, F.; Keane, E.; Macquart, J.-P.; Möller, A.; Onken, C. A.; Osłowski, S.; Parthasarathy, A.; Plant, K.; Ravi, V.; Shannon, R.; Tucker, B. E.; Venkatraman Krishnan, V.; Wolf, C.

    2018-05-01

    We report a new Fast Radio Burst (FRB) discovered in real-time as part of the UTMOST project at the Molonglo Observatory Synthesis Radio Telescope (MOST). FRB170827 is the first detected with our low-latency ( 20 ± 7 Jy ms, and is narrow, with a width of ˜ 400 μs at 10% of its maximum amplitude. However, the burst shows three temporal components, the narrowest of which is ˜ 30 μs, and a scattering timescale of 4.1 ± 2.7 μs. The FRB shows spectral modulations on frequency scales of 1.5 MHz and 0.1 MHz. Both are prominent in the dynamic spectrum, which shows a very bright region of emission between 841 and 843 MHz, and weaker, patchy emission across the entire band. We show the fine spectral structure could arise in the FRB host galaxy, or its immediate vicinity.

  2. A method for accurate detection of genomic microdeletions using real-time quantitative PCR

    Directory of Open Access Journals (Sweden)

    Bassett Anne S

    2005-12-01

    Full Text Available Abstract Background Quantitative Polymerase Chain Reaction (qPCR is a well-established method for quantifying levels of gene expression, but has not been routinely applied to the detection of constitutional copy number alterations of human genomic DNA. Microdeletions or microduplications of the human genome are associated with a variety of genetic disorders. Although, clinical laboratories routinely use fluorescence in situ hybridization (FISH to identify such cryptic genomic alterations, there remains a significant number of individuals in which constitutional genomic imbalance is suspected, based on clinical parameters, but cannot be readily detected using current cytogenetic techniques. Results In this study, a novel application for real-time qPCR is presented that can be used to reproducibly detect chromosomal microdeletions and microduplications. This approach was applied to DNA from a series of patient samples and controls to validate genomic copy number alteration at cytoband 22q11. The study group comprised 12 patients with clinical symptoms of chromosome 22q11 deletion syndrome (22q11DS, 1 patient trisomic for 22q11 and 4 normal controls. 6 of the patients (group 1 had known hemizygous deletions, as detected by standard diagnostic FISH, whilst the remaining 6 patients (group 2 were classified as 22q11DS negative using the clinical FISH assay. Screening of the patients and controls with a set of 10 real time qPCR primers, spanning the 22q11.2-deleted region and flanking sequence, confirmed the FISH assay results for all patients with 100% concordance. Moreover, this qPCR enabled a refinement of the region of deletion at 22q11. Analysis of DNA from chromosome 22 trisomic sample demonstrated genomic duplication within 22q11. Conclusion In this paper we present a qPCR approach for the detection of chromosomal microdeletions and microduplications. The strategic use of in silico modelling for qPCR primer design to avoid regions of repetitive

  3. A MIQE-compliant real-time PCR assay for Aspergillus detection.

    Directory of Open Access Journals (Sweden)

    Gemma L Johnson

    Full Text Available The polymerase chain reaction (PCR is widely used as a diagnostic tool in clinical laboratories and is particularly effective for detecting and identifying infectious agents for which routine culture and microscopy methods are inadequate. Invasive fungal disease (IFD is a major cause of morbidity and mortality in immunosuppressed patients, and optimal diagnostic criteria are contentious. Although PCR-based methods have long been used for the diagnosis of invasive aspergillosis (IA, variable performance in clinical practice has limited their value. This shortcoming is a consequence of differing sample selection, collection and preparation protocols coupled with a lack of standardisation of the PCR itself. Furthermore, it has become clear that the performance of PCR-based assays in general is compromised by the inadequacy of experimental controls, insufficient optimisation of assay performance as well as lack of transparency in reporting experimental details. The recently published "Minimum Information for the publication of real-time Quantitative PCR Experiments" (MIQE guidelines provide a blueprint for good PCR assay design and unambiguous reporting of experimental detail and results. We report the first real-time quantitative PCR (qPCR assay targeting Aspergillus species that has been designed, optimised and validated in strict compliance with the MIQE guidelines. The hydrolysis probe-based assay, designed to target the 18S rRNA DNA sequence of Aspergillus species, has an efficiency of 100% (range 95-107%, a dynamic range of at least six orders of magnitude and limits of quantification and detection of 6 and 0.6 Aspergillus fumigatus genomes, respectively. It does not amplify Candida, Scedosporium, Fusarium or Rhizopus species and its clinical sensitivity is demonstrated in histological material from proven IA cases, as well as concordant PCR and galactomannan data in matched broncho-alveolar lavage and blood samples. The robustness

  4. Quantitative detection of Campylobacter jejuni on fresh chicken carcasses by real-time PCR.

    Science.gov (United States)

    Rönner, Anna-Clara; Lindmark, Hans

    2007-06-01

    Campylobacter jejuni infection is a significant cause of foodborne gastroenteritis worldwide. Consumption and handling of poultry products is believed to be the primary risk factor for campylobacteriosis. Risk assessments require quantitative data, and C. jejuni is enumerated usually by direct plating, which sometimes allows growth of non-Campylobacter bacteria. The objective of the present study was to develop a quantitative real-time PCR method (q-PCR) for enumerating C. jejuni in chicken rinse without a culturing step. The procedure to obtain the template for the PCR assay involved (i) filtration of 10 ml of chicken rinse, (ii) centrifugation of the sample, and (iii) total DNA extraction from the pellet obtained using a commercial DNA extraction kit. The detection limit of the method was comparable to that for plating 100 microl of chicken rinse on modified charcoal cefoperazone deoxycholate agar, and the detection limit could be further improved 10-fold by concentrating the DNA eluate by ethanol precipitation. A close correlation for spiked chicken rinse was obtained for the results of the quantitative real-time PCR method and direct plating (r = 0.99). The coefficient of correlation for the methods was 0.87 when samples from chicken carcasses on the slaughter line were analyzed, whereas a lower correlation (r = 0.76) was obtained when samples from retail carcasses were analyzed. Greater variation in the proportion of dead and/or viable but not culturable Campylobacter types in the retail samples may explain the decreased correlation between the methods. Overall, the new method is simple and fast and the results obtained are closely correlated with those for direct plating for samples containing a low proportion of dead Campylobacter cells.

  5. Real-time in vivo diagnosis of laryngeal carcinoma with rapid fiber-optic Raman spectroscopy

    Science.gov (United States)

    Lin, Kan; Zheng, Wei; Lim, Chwee Ming; Huang, Zhiwei

    2016-01-01

    We assess the clinical utility of a unique simultaneous fingerprint (FP) (i.e., 800-1800 cm−1) and high-wavenumber (HW) (i.e., 2800-3600 cm−1) fiber-optic Raman spectroscopy for in vivo diagnosis of laryngeal cancer at endoscopy. A total of 2124 high-quality in vivo FP/HW Raman spectra (normal = 1321; cancer = 581) were acquired from 101 tissue sites (normal = 71; cancer = 30) of 60 patients (normal = 44; cancer = 16) undergoing routine endoscopic examination. FP/HW Raman spectra differ significantly between normal and cancerous laryngeal tissue that could be attributed to changes of proteins, lipids, nucleic acids, and the bound water content in the larynx. Partial least squares-discriminant analysis and leave-one tissue site-out, cross-validation were employed on the in vivo FP/HW tissue Raman spectra acquired, yielding a diagnostic accuracy of 91.1% (sensitivity: 93.3% (28/30); specificity: 90.1% (64/71)) for laryngeal cancer identification, which is superior to using either FP (accuracy: 86.1%; sensitivity: 86.7% (26/30); specificity: 85.9% (61/71)) or HW (accuracy: 84.2%; sensitivity: 76.7% (23/30); specificity: 87.3% (62/71)) Raman technique alone. Further receiver operating characteristic analysis reconfirms the best performance of the simultaneous FP/HW Raman technique for laryngeal cancer diagnosis. We demonstrate for the first time that the simultaneous FP/HW Raman spectroscopy technique can be used for improving real-time in vivo diagnosis of laryngeal carcinoma during endoscopic examination. PMID:27699131

  6. An immunomagnetic separation-real-time PCR system for the detection of Alicyclobacillus acidoterrestris in fruit products.

    Science.gov (United States)

    Wang, Zhouli; Cai, Rui; Yuan, Yahong; Niu, Chen; Hu, Zhongqiu; Yue, Tianli

    2014-04-03

    Alicyclobacillus acidoterrestris is the most important spoilage species within the Alicyclobacillus genus and has become a major issue in the pasteurized fruit juice industry. The aim of this study was to develop a method combining immunomagnetic separation (IMS) with real-time PCR system (IMS-PCR) for rapid and specific detection of A. acidoterrestris in fruit products. A real-time PCR with the TaqMan system was designed to target the 16S rDNA genes with specific primer and probe set. The specificity of the assay was confirmed using 9 A. acidoterrestris strains and 21 non-A. acidoterrestris strains. The results indicated that no combination of the designed primers and probe was found in any Alicyclobacillus genus except A. acidoterrestris. The detection limit of the established IMS-PCR was less than 10CFU/mL and the testing process was accomplished in 2-3h. For the three types of samples (sterile water, apple juice and kiwi juice), the correlation coefficient of standard curves was greater than 0.991, and the calculated PCR efficiencies were from 108% to 109%. As compared with the standard culture method performed concurrently on the same set of samples, the sensitivity, specificity and accuracy of IMS-PCR for 196 naturally contaminated fruit products were 90.0%, 98.3% and 97.5%, respectively. The results exhibited that the proposed IMS-PCR method was effective for the rapid detection of A. acidoterrestris in fruit products. Copyright © 2014. Published by Elsevier B.V.

  7. Detection of Tumor Markers in Prostate Cancer and Comparison of Sensitivity between Real Time and Nested PCR

    OpenAIRE

    Matsuoka, Takayuki; Shigemura, Katsumi; Yamamichi, Fukashi; Fujisawa, Masato; Kawabata, Masato; Shirakawa, Toshiro

    2012-01-01

    The objective of this study is to investigate and compare the sensitivity in conventional PCR, quantitative real time PCR, nested PCR and western blots for detection of prostate cancer tumor markers using prostate cancer (PCa) cells. We performed conventional PCR, quantitative real time PCR, nested PCR, and western blots using 5 kinds of PCa cells. Prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), and androgen receptor (AR) were compared for their detection sensitivi...

  8. Clinical evaluation of the Abbott RealTime MTB Assay for direct detection of Mycobacterium tuberculosis-complex from respiratory and non-respiratory samples.

    Science.gov (United States)

    Hinić, Vladimira; Feuz, Kinga; Turan, Selda; Berini, Andrea; Frei, Reno; Pfeifer, Karin; Goldenberger, Daniel

    2017-05-01

    Rapid and reliable diagnosis is crucial for correct management of tuberculosis. The Abbott RealTime MTB Assay represents a novel qualitative real-time PCR assay for direct detection of M. tuberculosis-complex (MTB) DNA from respiratory samples. The test targets two highly conserved sequences, the multi-copy insertion element IS6110 and the protein antigen B (PAB) gene of MTB, allowing even the detection of IS6610-deficient strains. We evaluated this commercial diagnostic test by analyzing 200 respiratory and, for the first time, 87 non-respiratory clinical specimens from our tertiary care institution and compared its results to our IS6110-based in-house real-time PCR for MTB as well as MTB culture. Overall sensitivity for Abbott RealTime MTB was 100% (19/19) in smear positive and 87.5% (7/8) in smear negative specimens, while the specificity of the assay was 100% (260/260). For both non-respiratory smear positive and smear negative specimens Abbott RealTime MTB tests showed 100% (8/8) sensitivity and 100% (8/8) specificity. Cycle threshold (Ct) value analysis of 16 MTB positive samples showed a slightly higher Ct value of the Abbott RealTime MTB test compared to our in-house MTB assay (mean delta Ct = 2.55). In conclusion, the performance of the new Abbott RealTime MTB Assay was highly similar to culture and in-house MTB PCR. We document successful analysis of 87 non-respiratory samples with the highly automated Abbott RealTime MTB test with no inhibition observed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Improvement of a picking algorithm real-time P-wave detection by kurtosis

    Science.gov (United States)

    Ishida, H.; Yamada, M.

    2016-12-01

    Earthquake early warning (EEW) requires fast and accurate P-wave detection. The current EEW system in Japan uses the STA/LTAalgorithm (Allen, 1978) to detect P-wave arrival.However, some stations did not trigger during the 2011 Great Tohoku Earthquake due to the emergent onset. In addition, accuracy of the P-wave detection is very important: on August 1, 2016, the EEW issued a false alarm with M9 in Tokyo region due to a thunder noise.To solve these problems, we use a P-wave detection method using kurtosis statistics. It detects the change of statistic distribution of the waveform amplitude. This method was recently developed (Saragiotis et al., 2002) and used for off-line analysis such as making seismic catalogs. To apply this method for EEW, we need to remove an acausal calculation and enable a real-time processing. Here, we propose a real-time P-wave detection method using kurtosis statistics with a noise filter.To avoid false triggering by a noise, we incorporated a simple filter to classify seismic signal and noise. Following Kong et al. (2016), we used the interquartilerange and zero cross rate for the classification. The interquartile range is an amplitude measure that is equal to the middle 50% of amplitude in a certain time window. The zero cross rate is a simple frequency measure that counts the number of times that the signal crosses baseline zero. A discriminant function including these measures was constructed by the linear discriminant analysis.To test this kurtosis method, we used strong motion records for 62 earthquakes between April, 2005 and July, 2015, which recorded the seismic intensity greater equal to 6 lower in the JMA intensity scale. The records with hypocentral distance picks. It shows that the median error is 0.13 sec and 0.035 sec for STA/LTA and kurtosis method. The kurtosis method tends to be more sensitive to small changes in amplitude.Our approach will contribute to improve the accuracy of source location determination of

  10. Real-time bicycle detection at signalized intersections using thermal imaging technology

    Science.gov (United States)

    Collaert, Robin

    2013-02-01

    More and more governments and authorities around the world are promoting the use of bicycles in cities, as this is healthy for the bicyclist and improves the quality of life in general. Safety and efficiency of bicyclists has become a major focus. To achieve this, there is a need for a smarter approach towards the control of signalized intersections. Various traditional detection technologies, such as video, microwave radar and electromagnetic loops, can be used to detect vehicles at signalized intersections, but none of these can consistently separate bikes from other traffic, day and night and in various weather conditions. As bikes should get a higher priority and also require longer green time to safely cross the signalized intersection, traffic managers are looking for alternative detection systems that can make the distinction between bicycles and other vehicles near the stop bar. In this paper, the drawbacks of a video-based approach are presented, next to the benefits of a thermal-video-based approach for vehicle presence detection with separation of bicycles. Also, the specific technical challenges are highlighted in developing a system that combines thermal image capturing, image processing and output triggering to the traffic light controller in near real-time and in a single housing.

  11. A Highly Sensitive Chemiluminometric Assay for Real-Time Detection of Biological Hydrogen Peroxide Formation.

    Science.gov (United States)

    Zhu, Hong; Jia, Zhenquan; Trush, Michael A; Li, Y Robert

    2016-05-01

    Hydrogen peroxide (H 2 O 2 ) is a major reactive oxygen species (ROS) produced by various cellular sources, especially mitochondria. At high levels, H 2 O 2 causes oxidative stress, leading to cell injury, whereas at low concentrations, this ROS acts as an important second messenger to participate in cellular redox signaling. Detection and measurement of the levels or rates of production of cellular H 2 O 2 are instrumental in studying the biological effects of this major ROS. While a number of assays have been developed over the past decades for detecting and/or quantifying biological H 2 O 2 formation, none has been shown to be perfect. Perhaps there is no perfect assay for sensitively and accurately quantifying H 2 O 2 as well as other ROS in cells, wherein numerous potential reactants are present to interfere with the reliable measurement of the specific ROS. In this context, each assay has its own advantages and intrinsic limitations. This article describes a highly sensitive assay for real-time detection of H 2 O 2 formation in cultured cells and isolated mitochondria. This assay is based on the luminol/horseradish peroxidase-dependent chemiluminescence that is inhibitable by catalase. The article discusses the usefulness and shortcomings of this chemiluminometric assay in detecting biological H 2 O 2 formation induced by beta-lapachone redox cycling with both cells and isolated mitochondria.

  12. Real time testing of intelligent relays for synchronous distributed generation islanding detection

    Science.gov (United States)

    Zhuang, Davy

    As electric power systems continue to grow to meet ever-increasing energy demand, their security, reliability, and sustainability requirements also become more stringent. The deployment of distributed energy resources (DER), including generation and storage, in conventional passive distribution feeders, gives rise to integration problems involving protection and unintentional islanding. Distributed generators need to be islanded for safety reasons when disconnected or isolated from the main feeder as distributed generator islanding may create hazards to utility and third-party personnel, and possibly damage the distribution system infrastructure, including the distributed generators. This thesis compares several key performance indicators of a newly developed intelligent islanding detection relay, against islanding detection devices currently used by the industry. The intelligent relay employs multivariable analysis and data mining methods to arrive at decision trees that contain both the protection handles and the settings. A test methodology is developed to assess the performance of these intelligent relays on a real time simulation environment using a generic model based on a real-life distribution feeder. The methodology demonstrates the applicability and potential advantages of the intelligent relay, by running a large number of tests, reflecting a multitude of system operating conditions. The testing indicates that the intelligent relay often outperforms frequency, voltage and rate of change of frequency relays currently used for islanding detection, while respecting the islanding detection time constraints imposed by standing distributed generator interconnection guidelines.

  13. A Wireless Sensor System for Real-Time Monitoring and Fault Detection of Motor Arrays.

    Science.gov (United States)

    Medina-García, Jonathan; Sánchez-Rodríguez, Trinidad; Galán, Juan Antonio Gómez; Delgado, Aránzazu; Gómez-Bravo, Fernando; Jiménez, Raúl

    2017-02-25

    This paper presents a wireless fault detection system for industrial motors that combines vibration, motor current and temperature analysis, thus improving the detection of mechanical faults. The design also considers the time of detection and further possible actions, which are also important for the early detection of possible malfunctions, and thus for avoiding irreversible damage to the motor. The remote motor condition monitoring is implemented through a wireless sensor network (WSN) based on the IEEE 802.15.4 standard. The deployed network uses the beacon-enabled mode to synchronize several sensor nodes with the coordinator node, and the guaranteed time slot mechanism provides data monitoring with a predetermined latency. A graphic user interface offers remote access to motor conditions and real-time monitoring of several parameters. The developed wireless sensor node exhibits very low power consumption since it has been optimized both in terms of hardware and software. The result is a low cost, highly reliable and compact design, achieving a high degree of autonomy of more than two years with just one 3.3 V/2600 mAh battery. Laboratory and field tests confirm the feasibility of the wireless system.

  14. Evaluation of different enrichment methods for pathogenic Yersinia species detection by real time PCR

    Science.gov (United States)

    2014-01-01

    Background Yersiniosis is a zoonotic disease reported worldwide. Culture and PCR based protocols are the most common used methods for detection of pathogenic Yersinia species in animal samples. PCR sensitivity could be increased by an initial enrichment step. This step is particularly useful in surveillance programs, where PCR is applied to samples from asymptomatic animals. The aim of this study was to evaluate the improvement in pathogenic Yersinia species detection using a suitable enrichment method prior to the real time PCR (rtPCR). Nine different enrichment protocols were evaluated including six different broth mediums (CASO, ITC, PSB, PBS, PBSMSB and PBSSSB). Results The analysis of variance showed significant differences in Yersinia detection by rtPCR according to the enrichment protocol used. These differences were higher for Y. pseudotuberculosis than for Y. enterocolitica. In general, samples incubated at lower temperatures yielded the highest detection rates. The best results were obtained with PBSMSB and PBS2. Application of PBSMSB protocol to free-ranging wild board samples improved the detection of Y. enterocolitica by 21.2% when compared with direct rtPCR. Y. pseudotuberculosis detection was improved by 10.6% when results obtained by direct rtPCR and by PBSMSB enrichment before rtPCR were analyzed in combination. Conclusions The data obtained in the present study indicate a difference in Yersinia detection by rtPCR related to the enrichment protocol used, being PBSMSB enrichment during 15 days at 4°C and PBS during 7 days at 4°C the most efficient. The use of direct rtPCR in combination with PBSMSB enrichment prior to rtPCR resulted in an improvement in the detection rates of pathogenic Yersinia in wild boar and could be useful for application in other animal samples. PMID:25168886

  15. Quantitative detection of the potato cyst nematode, Globodera pallida, and the beet cyst nematode, Heterodera schachtii, using Real-Time PCR with SYBR green I dye.

    Science.gov (United States)

    Madani, Mehrdad; Subbotin, Sergei A; Moens, Maurice

    2005-04-01

    The potato cyst nematode Globodera pallida and the beet cyst nematode Heterodera schachtii are major nematode pests in world agriculture. Precise identification and knowledge about the number of nematodes in field soil are necessary to develop effective integrated pest control. Here we report the results of the Real-Time PCR assay for the rapid detection and quantification of G. pallida and H. schachtii. Using species specific primers and SYBR green I dye, we were able to detect a single second stage juvenile of cyst forming nematodes in samples. The specificity of the reaction was confirmed by the lack of amplification of DNAs from other Heterodera or Globodera species. Validation tests showed a rather high correlation between real numbers of second stage juveniles in a sample and expected numbers detected by Real-Time PCR. Reasons for observed differences in sensitivity and reliability of quantification detection for two species as well as other problems of Real-Time PCR are discussed. The Real-Time PCR assay with SYBR green I dye targeting fragments of the ITS-rDNA provided a sensitive means for the rapid and simultaneous detection and quantification of juveniles of these pests.

  16. Real-Time Detection Methods to Monitor TRU Compositions in UREX+Process Streams

    Energy Technology Data Exchange (ETDEWEB)

    McDeavitt, Sean; Charlton, William; Indacochea, J Ernesto; taleyarkhan, Rusi; Pereira, Candido

    2013-03-01

    The U.S. Department of Energy has developed advanced methods for reprocessing spent nuclear fuel. The majority of this development was accomplished under the Advanced Fuel Cycle Initiative (AFCI), building on the strong legacy of process development R&D over the past 50 years. The most prominent processing method under development is named UREX+. The name refers to a family of processing methods that begin with the Uranium Extraction (UREX) process and incorporate a variety of other methods to separate uranium, selected fission products, and the transuranic (TRU) isotopes from dissolved spent nuclear fuel. It is important to consider issues such as safeguards strategies and materials control and accountability methods. Monitoring of higher actinides during aqueous separations is a critical research area. By providing on-line materials accountability for the processes, covert diversion of the materials streams becomes much more difficult. The importance of the nuclear fuel cycle continues to rise on national and international agendas. The U.S. Department of Energy is evaluating and developing advanced methods for safeguarding nuclear materials along with instrumentation in various stages of the fuel cycle, especially in material balance areas (MBAs) and during reprocessing of used nuclear fuel. One of the challenges related to the implementation of any type of MBA and/or reprocessing technology (e.g., PUREX or UREX) is the real-time quantification and control of the transuranic (TRU) isotopes as they move through the process. Monitoring of higher actinides from their neutron emission (including multiplicity) and alpha signatures during transit in MBAs and in aqueous separations is a critical research area. By providing on-line real-time materials accountability, diversion of the materials becomes much more difficult. The objective of this consortium was to develop real time detection methods to monitor the efficacy of the UREX+ process and to safeguard the separated

  17. Development of a sensitive real-time PCR for simultaneous detection and subtyping of influenza A and B viruses

    Directory of Open Access Journals (Sweden)

    Daniela Amicizia

    2005-03-01

    Full Text Available

    A new real-time PCR assay, using melting curve analysis, was developed for the rapid and reliable detection and sub-typing of influenza A and B.

    In order to evaluate it’s specificity, cell culture surnatants positive for Respiratory Syncytial Virus, Parainfluenza Viruses 1, 2 and 3, Measles Virus, Influenza A (to evaluate Influenza B primer and B (to evaluate Influenza A primer were tested and all of the results were negative.

    A series of Influenza A and B cell culture-grown viruses were diluted in virus transport medium, titrated and tested to determine the analytical sensibility which equated to 0.64, 0.026, 0.64, 0.62 PFU for A/H1N1, A/H3N2, Victoria-like and Yamagata-like B viruses, respectively. Twenty-five specimens, collected during the 2001/02 and 2002/03 seasons, which were positive for A/H1N1 (n = 7, A/H3N2 (n = 10, B Victoria-lineage (n = 5 and B Yamagata-lineage (n = 3, were tested in order to evaluate the assay’s clinical sensitivity, all of the results were positive.

    The new real-time PCR appears to be a suitable tool for virological surveillance and the diagnosis of respiratory infections.

  18. Porous Silicon-Based Biosensors: Towards Real-Time Optical Detection of Target Bacteria in the Food Industry.

    Science.gov (United States)

    Massad-Ivanir, Naama; Shtenberg, Giorgi; Raz, Nitzan; Gazenbeek, Christel; Budding, Dries; Bos, Martine P; Segal, Ester

    2016-11-30

    Rapid detection of target bacteria is crucial to provide a safe food supply and to prevent foodborne diseases. Herein, we present an optical biosensor for identification and quantification of Escherichia coli (E. coli, used as a model indicator bacteria species) in complex food industry process water. The biosensor is based on a nanostructured, oxidized porous silicon (PSi) thin film which is functionalized with specific antibodies against E. coli. The biosensors were exposed to water samples collected directly from process lines of fresh-cut produce and their reflectivity spectra were collected in real time. Process water were characterized by complex natural micro-flora (microbial load of >10 7  cell/mL), in addition to soil particles and plant cell debris. We show that process water spiked with culture-grown E. coli, induces robust and predictable changes in the thin-film optical interference spectrum of the biosensor. The latter is ascribed to highly specific capture of the target cells onto the biosensor surface, as confirmed by real-time polymerase chain reaction (PCR). The biosensors were capable of selectively identifying and quantifying the target cells, while the target cell concentration is orders of magnitude lower than that of other bacterial species, without any pre-enrichment or prior processing steps.

  19. A new comparison of hyperspectral anomaly detection algorithms for real-time applications

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    Díaz, María.; López, Sebastián.; Sarmiento, Roberto

    2016-10-01

    Due to the high spectral resolution that remotely sensed hyperspectral images provide, there has been an increasing interest in anomaly detection. The aim of anomaly detection is to stand over pixels whose spectral signature differs significantly from the background spectra. Basically, anomaly detectors mark pixels with a certain score, considering as anomalies those whose scores are higher than a threshold. Receiver Operating Characteristic (ROC) curves have been widely used as an assessment measure in order to compare the performance of different algorithms. ROC curves are graphical plots which illustrate the trade- off between false positive and true positive rates. However, they are limited in order to make deep comparisons due to the fact that they discard relevant factors required in real-time applications such as run times, costs of misclassification and the competence to mark anomalies with high scores. This last fact is fundamental in anomaly detection in order to distinguish them easily from the background without any posterior processing. An extensive set of simulations have been made using different anomaly detection algorithms, comparing their performances and efficiencies using several extra metrics in order to complement ROC curves analysis. Results support our proposal and demonstrate that ROC curves do not provide a good visualization of detection performances for themselves. Moreover, a figure of merit has been proposed in this paper which encompasses in a single global metric all the measures yielded for the proposed additional metrics. Therefore, this figure, named Detection Efficiency (DE), takes into account several crucial types of performance assessment that ROC curves do not consider. Results demonstrate that algorithms with the best detection performances according to ROC curves do not have the highest DE values. Consequently, the recommendation of using extra measures to properly evaluate performances have been supported and justified by

  20. Real-time detection of metal ions using conjugated polymer composite papers.

    Science.gov (United States)

    Lee, Ji Eun; Shim, Hyeon Woo; Kwon, Oh Seok; Huh, Yang-Il; Yoon, Hyeonseok

    2014-09-21

    Cellulose, a natural polymeric material, has widespread technical applications because of its inherent structural rigidity and high surface area. As a conjugated polymer, polypyrrole shows practical potential for a diverse and promising range of future technologies. Here, we demonstrate a strategy for the real-time detection and removal of metal ions with polypyrrole/cellulose (PPCL) composite papers in solution. Simply, the conjugated polymer papers had different chemical/physical properties by applying different potentials to them, which resulted in differentiable response patterns and adsorption efficiencies for individual metal ions. First, large-area PPCL papers with a diameter of 5 cm were readily obtained via vapor deposition polymerization. The papers exhibited both mechanical flexibility and robustness, in which polypyrrole retained its redox property perfectly. The ability of the PPCL papers to recognize metal ions was examined in static and flow cells, in which real-time current change was monitored at five different applied potentials (+1, +0.5, 0, -0.5, and -1 V vs. Ag/AgCl). Distinguishable signals in the PPCL paper responses were observed for individual metal ions through principal component analysis. Particularly, the PPCL papers yielded unique signatures for three metal ions, Hg(ii), Ag(i), and Cr(iii), even in a real sample, groundwater. The sorption of metal ions by PPCL papers was examined in the flow system. The PPCL papers had a greatly superior adsorption efficiency for Hg(ii) compared to that of the other metal ions. With the strong demand for the development of inexpensive, flexible, light-weight, and environmentally friendly devices, the fascinating characteristics of these PPCL papers are likely to provide good opportunities for low-cost paper-based flexible or wearable devices.

  1. Towards Real-Time Facial Landmark Detection in Depth Data Using Auxiliary Information

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    Connah Kendrick

    2018-06-01

    Full Text Available Modern facial motion capture systems employ a two-pronged approach for capturing and rendering facial motion. Visual data (2D is used for tracking the facial features and predicting facial expression, whereas Depth (3D data is used to build a series of expressions on 3D face models. An issue with modern research approaches is the use of a single data stream that provides little indication of the 3D facial structure. We compare and analyse the performance of Convolutional Neural Networks (CNN using visual, Depth and merged data to identify facial features in real-time using a Depth sensor. First, we review the facial landmarking algorithms and its datasets for Depth data. We address the limitation of the current datasets by introducing the Kinect One Expression Dataset (KOED. Then, we propose the use of CNNs for the single data stream and merged data streams for facial landmark detection. We contribute to existing work by performing a full evaluation on which streams are the most effective for the field of facial landmarking. Furthermore, we improve upon the existing work by extending neural networks to predict into 3D landmarks in real-time with additional observations on the impact of using 2D landmarks as auxiliary information. We evaluate the performance by using Mean Square Error (MSE and Mean Average Error (MAE. We observe that the single data stream predicts accurate facial landmarks on Depth data when auxiliary information is used to train the network. The codes and dataset used in this paper will be made available.

  2. Rapid identification and quantification of Campylobacter coli and Campylobacter jejuni by real-time PCR in pure cultures and in complex samples

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    Denis Martine

    2011-05-01

    Full Text Available Abstract Background Campylobacter spp., especially Campylobacter jejuni (C. jejuni and Campylobacter coli (C. coli, are recognized as the leading human foodborne pathogens in developed countries. Livestock animals carrying Campylobacter pose an important risk for human contamination. Pigs are known to be frequently colonized with Campylobacter, especially C. coli, and to excrete high numbers of this pathogen in their faeces. Molecular tools, notably real-time PCR, provide an effective, rapid, and sensitive alternative to culture-based methods for the detection of C. coli and C. jejuni in various substrates. In order to serve as a diagnostic tool supporting Campylobacter epidemiology, we developed a quantitative real-time PCR method for species-specific detection and quantification of C. coli and C. jejuni directly in faecal, feed, and environmental samples. Results With a sensitivity of 10 genome copies and a linear range of seven to eight orders of magnitude, the C. coli and C. jejuni real-time PCR assays allowed a precise quantification of purified DNA from C. coli and C. jejuni. The assays were highly specific and showed a 6-log-linear dynamic range of quantification with a quantitative detection limit of approximately 2.5 × 102 CFU/g of faeces, 1.3 × 102 CFU/g of feed, and 1.0 × 103 CFU/m2 for the environmental samples. Compared to the results obtained by culture, both C. coli and C. jejuni real-time PCR assays exhibited a specificity of 96.2% with a kappa of 0.94 and 0.89 respectively. For faecal samples of experimentally infected pigs, the coefficients of correlation between the C. coli or C. jejuni real-time PCR assay and culture enumeration were R2 = 0.90 and R2 = 0.93 respectively. Conclusion The C. coli and C. jejuni real-time quantitative PCR assays developed in this study provide a method capable of directly detecting and quantifying C. coli and C. jejuni in faeces, feed, and environmental samples. These assays represent a new

  3. Improved quantification accuracy for duplex real-time PCR detection of genetically modified soybean and maize in heat processed foods

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    CHENG Fang

    2013-04-01

    Full Text Available Real-time PCR technique has been widely used in quantitative GMO detection in recent years.The accuracy of GMOs quantification based on the real-time PCR methods is still a difficult problem,especially for the quantification of high processed samples.To develop the suitable and accurate real-time PCR system for high processed GM samples,we made ameliorations to several real-time PCR parameters,including re-designed shorter target DNA fragment,similar lengths of amplified endogenous and exogenous gene targets,similar GC contents and melting temperatures of PCR primers and TaqMan probes.Also,one Heat-Treatment Processing Model (HTPM was established using soybean flour samples containing GM soybean GTS 40-3-2 to validate the effectiveness of the improved real-time PCR system.Tested results showed that the quantitative bias of GM content in heat processed samples were lowered using the new PCR system.The improved duplex real-time PCR was further validated using processed foods derived from GM soybean,and more accurate GM content values in these foods was also achieved.These results demonstrated that the improved duplex real-time PCR would be quite suitable in quantitative detection of high processed food products.

  4. A real-time PCR approach to detect predation on anchovy and sardine early life stages

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    Cuende, Elsa; Mendibil, Iñaki; Bachiller, Eneko; Álvarez, Paula; Cotano, Unai; Rodriguez-Ezpeleta, Naiara

    2017-12-01

    Recruitment of sardine (Sardina pilchardus Walbaum, 1792) and anchovy (Engraulis encrasicolus Linnaeus, 1758) is thought to be regulated by predation of their eggs and larvae. Predators of sardine and anchovy can be identified by visual taxonomic identification of stomach contents, but this method is time consuming, tedious and may underestimate predation, especially in small predators such as fish larvae. Alternatively, genetic tools may offer a more cost-effective and accurate alternative. Here, we have developed a multiplex real-time polymerase chain reaction (RT-PCR) assay based on TaqMan probes to simultaneously detect sardine and anchovy remains in gut contents of potential predators. The assay combines previously described and newly generated species-specific primers and probes for anchovy and sardine detection respectively, and allows the detection of 0,001 ng of target DNA (which corresponds to about one hundredth of the total DNA present in a single egg). We applied the method to candidate anchovy and sardine egg predators in the Bay of Biscay, Atlantic Mackerel (Scomber scombrus) larvae. Egg predation observed was limited primarily to those stations where sardine and/or anchovy eggs were present. Our developed assay offers a suitable tool to understand the effects of predation on the survival of anchovy and sardine early life stages.

  5. Real-Time Detection of Staphylococcus Aureus Using Whispering Gallery Mode Optical Microdisks

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    Hala Ghali

    2016-05-01

    Full Text Available Whispering Gallery Mode (WGM microresonators have recently been studied as a means to achieve real-time label-free detection of biological targets such as virus particles, specific DNA sequences, or proteins. Due to their high quality (Q factors, WGM resonators can be highly sensitive. A biosensor also needs to be selective, requiring proper functionalization of its surface with the appropriate ligand that will attach the biomolecule of interest. In this paper, WGM microdisks are used as biosensors for detection of Staphylococcus aureus. The microdisks are functionalized with LysK, a phage protein specific for staphylococci at the genus level. A binding event on the surface shifts the resonance peak of the microdisk resonator towards longer wavelengths. This reactive shift can be used to estimate the surface density of bacteria that bind to the surface of the resonator. The limit of detection of a microdisk with a Q-factor around 104 is on the order of 5 pg/mL, corresponding to 20 cells. No binding of Escherichia coli to the resonators is seen, supporting the specificity of the functionalization scheme.

  6. A real time QRS detection using delay-coordinate mapping for the microcontroller implementation.

    Science.gov (United States)

    Lee, Jeong-Whan; Kim, Kyeong-Seop; Lee, Bongsoo; Lee, Byungchae; Lee, Myoung-Ho

    2002-01-01

    In this article, we propose a new algorithm using the characteristics of reconstructed phase portraits by delay-coordinate mapping utilizing lag rotundity for a real-time detection of QRS complexes in ECG signals. In reconstructing phase portrait the mapping parameters, time delay, and mapping dimension play important roles in shaping of portraits drawn in a new dimensional space. Experimentally, the optimal mapping time delay for detection of QRS complexes turned out to be 20 ms. To explore the meaning of this time delay and the proper mapping dimension, we applied a fill factor, mutual information, and autocorrelation function algorithm that were generally used to analyze the chaotic characteristics of sampled signals. From these results, we could find the fact that the performance of our proposed algorithms relied mainly on the geometrical property such as an area of the reconstructed phase portrait. For the real application, we applied our algorithm for designing a small cardiac event recorder. This system was to record patients' ECG and R-R intervals for 1 h to investigate HRV characteristics of the patients who had vasovagal syncope symptom and for the evaluation, we implemented our algorithm in C language and applied to MIT/BIH arrhythmia database of 48 subjects. Our proposed algorithm achieved a 99.58% detection rate of QRS complexes.

  7. Detection of Yersinia Enterocolitica Species in Pig Tonsils and Raw Pork Meat by the Real-Time Pcr and Culture Methods.

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    Stachelska, M A

    2017-09-26

    The aim of the present study was to establish a rapid and accurate real-time PCR method to detect pathogenic Yersinia enterocolitica in pork. Yersinia enterocolitica is considered to be a crucial zoonosis, which can provoke diseases both in humans and animals. The classical culture methods designated to detect Y. enterocolitica species in food matrices are often very time-consuming. The chromosomal locus _tag CH49_3099 gene, that appears in pathogenic Y. enterocolitica strains, was applied as DNA target for the 5' nuclease PCR protocol. The probe was labelled at the 5' end with the fluorescent reporter dye (FAM) and at the 3' end with the quencher dye (TAMRA). The real-time PCR cycling parameters included 41 cycles. A Ct value which reached a value higher than 40 constituted a negative result. The developed for the needs of this study qualitative real-time PCR method appeared to give very specific and reliable results. The detection rate of locus _tag CH49_3099 - positive Y. enterocolitica in 150 pig tonsils was 85 % and 32 % with PCR and culture methods, respectively. Both the Real-time PCR results and culture method results were obtained from material that was enriched during overnight incubation. The subject of the study were also raw pork meat samples. Among 80 samples examined, 7 ones were positive when real-time PCR was applied, and 6 ones were positive when classical culture method was applied. The application of molecular techniques based on the analysis of DNA sequences such as the Real-time PCR enables to detect this pathogenic bacteria very rapidly and with higher specificity, sensitivity and reliability in comparison to classical culture methods.

  8. A Novel Real-Time DDoS Attack Detection Mechanism Based on MDRA Algorithm in Big Data

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    Bin Jia

    2016-01-01

    Full Text Available In the wake of the rapid development and wide application of information technology and Internet, our society has come into the information explosion era. Meanwhile, it brings in new and severe challenges to the field of network attack behavior detection due to the explosive growth and high complexity of network traffic. Therefore, an effective and efficient detection mechanism that can detect attack behavior from large scale of network traffic plays an important role. In this paper, we focus on how to distinguish the attack traffic from normal data flows in Big Data and propose a novel real-time DDoS attack detection mechanism based on Multivariate Dimensionality Reduction Analysis (MDRA. In this mechanism, we first reduce the dimensionality of multiple characteristic variables in a network traffic record by Principal Component Analysis (PCA. Then, we analyze the correlation of the lower dimensional variables. Finally, the attack traffic can be differentiated from the normal traffic by MDRA and Mahalanobis distance (MD. Compared with previous research methods, our experimental results show that higher precision rate is achieved and it approximates to 100% in True Negative Rate (TNR for detection; CPU computing time is one-eightieth and memory resource consumption is one-third of the previous detection method based on Multivariate Correlation Analysis (MCA; computing complexity is constant.

  9. Detection of 12 respiratory viruses by duplex real time PCR assays in respiratory samples.

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    Arvia, Rosaria; Corcioli, Fabiana; Ciccone, Nunziata; Della Malva, Nunzia; Azzi, Alberta

    2015-12-01

    Different viruses can be responsible for similar clinical manifestations of respiratory infections. Thus, the etiological diagnosis of respiratory viral diseases requires the detection of a large number of viruses. In this study, 6 duplex real-time PCR assays, using EvaGreen intercalating dye, were developed to detect 12 major viruses responsible for respiratory diseases: influenza A and B viruses, enteroviruses (including enterovirus spp, and rhinovirus spp), respiratory syncytial virus, human metapneumovirus, coronaviruses group I (of which CoV 229E and CoV NL63 are part) and II (including CoV OC43 and CoV HKU1), parainfluenza viruses type 1, 2, 3 and 4, human adenoviruses and human bocaviruses. The 2 target viruses of each duplex reaction were distinguishable by the melting temperatures of their amplicons. The 6 duplex real time PCR assays were applied for diagnostic purpose on 202 respiratory samples from 157 patients. One hundred fifty-seven samples were throat swabs and 45 were bronchoalveolar lavages. The results of the duplex PCR assays were confirmed by comparison with a commercial, validated, assay; in addition, the positive results were confirmed by sequencing. The analytical sensitivity of the duplex PCR assays varied from 10(3) copies/ml to 10(4) copies/ml. For parainfluenza virus 2 only it was 10(5) copies/ml. Seventy clinical samples (35%) from 55 patients (30 children and 25 adults) were positive for 1 or more viruses. In adult patients, influenza A virus was the most frequently detected respiratory virus followed by rhinoviruses. In contrast, respiratory syncytial virus was the most common virus in children, followed by enteroviruses, influenza A virus and coronavirus NL63. The small number of samples/patients does not allow us to draw any epidemiological conclusion. Altogether, the results of this study indicate that the 6 duplex PCR assays described in this study are sensitive, specific and cost-effective. Thus, this assay could be

  10. Rapid, actionable diagnosis of urban epidemic leptospirosis using a pathogenic Leptospira lipL32-based real-time PCR assay.

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    Irina N Riediger

    2017-09-01

    Full Text Available With a conservatively estimated 1 million cases of leptospirosis worldwide and a 5-10% fatality rate, the rapid diagnosis of leptospirosis leading to effective clinical and public health decision making is of high importance, and yet remains a challenge.Based on parallel, population-based studies in two leptospirosis-endemic regions in Brazil, a real-time PCR assay which detects lipL32, a gene specifically present in pathogenic Leptospira, was assessed for the diagnostic effectiveness and accuracy. Patients identified by active hospital-based surveillance in Salvador and Curitiba during large urban leptospirosis epidemics were tested. Real-time PCR reactions were performed with DNA-extracted samples obtained from 127 confirmed and 23 unconfirmed cases suspected of leptospirosis, 122 patients with an acute febrile illness other than leptospirosis, and 60 healthy blood donors.The PCR assay had a limit of detection of 280 Leptospira genomic equivalents/mL. Sensitivity for confirmed cases was 61% for whole blood and 29% for serum samples. Sensitivity was higher (86% for samples collected within the first 6 days after onset of illness compared to those collected after 7 days (34%. The real-time PCR assay was able to detect leptospiral DNA in blood from 56% of serological non-confirmed cases. The overall specificity of the assay was 99%.These findings indicate that real-time PCR may be a reliable tool for early diagnosis of leptospirosis, which is decisive for clinical management of severe and life-threatening cases and for public health decision making.

  11. Rapid, actionable diagnosis of urban epidemic leptospirosis using a pathogenic Leptospira lipL32-based real-time PCR assay.

    Science.gov (United States)

    Riediger, Irina N; Stoddard, Robyn A; Ribeiro, Guilherme S; Nakatani, Sueli M; Moreira, Suzana D R; Skraba, Irene; Biondo, Alexander W; Reis, Mitermayer G; Hoffmaster, Alex R; Vinetz, Joseph M; Ko, Albert I; Wunder, Elsio A

    2017-09-01

    With a conservatively estimated 1 million cases of leptospirosis worldwide and a 5-10% fatality rate, the rapid diagnosis of leptospirosis leading to effective clinical and public health decision making is of high importance, and yet remains a challenge. Based on parallel, population-based studies in two leptospirosis-endemic regions in Brazil, a real-time PCR assay which detects lipL32, a gene specifically present in pathogenic Leptospira, was assessed for the diagnostic effectiveness and accuracy. Patients identified by active hospital-based surveillance in Salvador and Curitiba during large urban leptospirosis epidemics were tested. Real-time PCR reactions were performed with DNA-extracted samples obtained from 127 confirmed and 23 unconfirmed cases suspected of leptospirosis, 122 patients with an acute febrile illness other than leptospirosis, and 60 healthy blood donors. The PCR assay had a limit of detection of 280 Leptospira genomic equivalents/mL. Sensitivity for confirmed cases was 61% for whole blood and 29% for serum samples. Sensitivity was higher (86%) for samples collected within the first 6 days after onset of illness compared to those collected after 7 days (34%). The real-time PCR assay was able to detect leptospiral DNA in blood from 56% of serological non-confirmed cases. The overall specificity of the assay was 99%. These findings indicate that real-time PCR may be a reliable tool for early diagnosis of leptospirosis, which is decisive for clinical management of severe and life-threatening cases and for public health decision making.

  12. Detection and enumeration of Salmonella enteritidis in homemade ice cream associated with an outbreak: comparison of conventional and real-time PCR methods.

    Science.gov (United States)

    Seo, K H; Valentin-Bon, I E; Brackett, R E

    2006-03-01

    Salmonellosis caused by Salmonella Enteritidis (SE) is a significant cause of foodborne illnesses in the United States. Consumption of undercooked eggs and egg-containing products has been the primary risk factor for the disease. The importance of the bacterial enumeration technique has been enormously stressed because of the quantitative risk analysis of SE in shell eggs. Traditional enumeration methods mainly depend on slow and tedious most-probable-number (MPN) methods. Therefore, specific, sensitive, and rapid methods for SE quantitation are needed to collect sufficient data for risk assessment and food safety policy development. We previously developed a real-time quantitative PCR assay for the direct detection and enumeration of SE and, in this study, applied it to naturally contaminated ice cream samples with and without enrichment. The detection limit of the real-time PCR assay was determined with artificially inoculated ice cream. When applied to the direct detection and quantification of SE in ice cream, the real-time PCR assay was as sensitive as the conventional plate count method in frequency of detection. However, populations of SE derived from real-time quantitative PCR were approximately 1 log higher than provided by MPN and CFU values obtained by conventional culture methods. The detection and enumeration of SE in naturally contaminated ice cream can be completed in 3 h by this real-time PCR method, whereas the cultural enrichment method requires 5 to 7 days. A commercial immunoassay for the specific detection of SE was also included in the study. The real-time PCR assay proved to be a valuable tool that may be useful to the food industry in monitoring its processes to improve product quality and safety.

  13. Rapid and minimum invasive functional brain mapping by real-time visualization of high gamma activity during awake craniotomy.

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    Ogawa, Hiroshi; Kamada, Kyousuke; Kapeller, Christoph; Hiroshima, Satoru; Prueckl, Robert; Guger, Christoph

    2014-11-01

    Electrocortical stimulation (ECS) is the gold standard for functional brain mapping during an awake craniotomy. The critical issue is to set aside enough time to identify eloquent cortices by ECS. High gamma activity (HGA) ranging between 80 and 120 Hz on electrocorticogram is assumed to reflect localized cortical processing. In this report, we used real-time HGA mapping and functional neuronavigation integrated with functional magnetic resonance imaging (fMRI) for rapid and reliable identification of motor and language functions. Four patients with intra-axial tumors in their dominant hemisphere underwent preoperative fMRI and lesion resection with an awake craniotomy. All patients showed significant fMRI activation evoked by motor and language tasks. During the craniotomy, we recorded electrocorticogram activity by placing subdural grids directly on the exposed brain surface. Each patient performed motor and language tasks and demonstrated real-time HGA dynamics in hand motor areas and parts of the inferior frontal gyrus. Sensitivity and specificity of HGA mapping were 100% compared with ECS mapping in the frontal lobe, which suggested HGA mapping precisely indicated eloquent cortices. We found different HGA dynamics of language tasks in frontal and temporal regions. Specificities of the motor and language-fMRI did not reach 85%. The results of HGA mapping was mostly consistent with those of ECS mapping, although fMRI tended to overestimate functional areas. This novel technique enables rapid and accurate identification of motor and frontal language areas. Furthermore, real-time HGA mapping sheds light on underlying physiological mechanisms related to human brain functions. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. Using quantitative real-time PCR to detect chimeras in transgenic tobacco and apricot and to monitor their dissociation

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    Burgos Lorenzo

    2010-07-01

    Full Text Available Abstract Background The routine generation of transgenic plants involves analysis of transgene integration into the host genome by means of Southern blotting. However, this technique cannot distinguish between uniformly transformed tissues and the presence of a mixture of transgenic and non-transgenic cells in the same tissue. On the other hand, the use of reporter genes often fails to accurately detect chimerical tissues because their expression can be affected by several factors, including gene silencing and plant development. So, new approaches based on the quantification of the amount of the transgene are needed urgently. Results We show here that chimeras are a very frequent phenomenon observed after regenerating transgenic plants. Spatial and temporal analyses of transformed tobacco and apricot plants with a quantitative, real-time PCR amplification of the neomycin phosphotransferase (nptII transgene as well as of an internal control (β-actin, used to normalise the amount of target DNA at each reaction, allowed detection of chimeras at unexpected rates. The amount of the nptII transgene differed greatly along with the sub-cultivation period of these plants and was dependent on the localisation of the analysed leaves; being higher in roots and basal leaves, while in the apical leaves it remained at lower levels. These data demonstrate that, unlike the use of the gus marker gene, real-time PCR is a powerful tool for detection of chimeras. Although some authors have proposed a consistent, positive Southern analysis as an alternative methodology for monitoring the dissociation of chimeras, our data show that it does not provide enough proof of uniform transformation. In this work, however, real-time PCR was applied successfully to monitor the dissociation of chimeras in tobacco plants and apricot callus. Conclusions We have developed a rapid and reliable method to detect and estimate the level of chimeras in transgenic tobacco and apricot

  15. Real-time monitoring system for elderly people in detecting falling movement using accelerometer and gyroscope

    Science.gov (United States)

    Siregar, B.; Andayani, U.; Bahri, R. P.; Seniman; Fahmi, F.

    2018-03-01

    Most of the elderly people is experiencing a decrease in physical quality, especially the weakness in the legs. This will cause elderly easy to fall and can have a serious impact on their health if not getting help very quickly. It is, therefore, necessary to take immediate action against the falling cases experienced by the elderly. One such action is by developing supervision and detecting of falling movements in real-time, which is then the connection to a member of the family. In this research, we used Arduino Uno as a microcontroller, sensor accelerometer, and gyroscope that serves to measure falling movement of the elderly person and supported by GPS technology Ublox Neo 6M to provide information about coordinates. The result was the high accuracy of delivering notification data to server and accuracy of data delivery to family notification equal to 93,75%. The system successfully detects the direction of falling: forward, backward, left or right and able to distinguish between unintentional falling and conscious falling like a bow or prostrate position.

  16. Powerful conveyer belt real-time online detection system based on x-ray

    Science.gov (United States)

    Rong, Feng; Miao, Chang-yun; Meng, Wei

    2009-07-01

    The powerful conveyer belt is widely used in the mine, dock, and so on. After used for a long time, internal steel rope of the conveyor belt may fracture, rust, joints moving, and so on .This would bring potential safety problems. A kind of detection system based on x-ray is designed in this paper. Linear array detector (LDA) is used. LDA cost is low, response fast; technology mature .Output charge of LDA is transformed into differential voltage signal by amplifier. This kind of signal have great ability of anti-noise, is suitable for long-distance transmission. The processor is FPGA. A IP core control 4-channel A/D convertor, achieve parallel output data collection. Soft-core processor MicroBlaze which process tcp/ip protocol is embedded in FPGA. Sampling data are transferred to a computer via Ethernet. In order to improve the image quality, algorithm of getting rid of noise from the measurement result and taking gain normalization for pixel value is studied and designed. Experiments show that this system work well, can real-time online detect conveyor belt of width of 2.0m and speed of 5 m/s, does not affect the production. Image is clear, visual and can easily judge the situation of conveyor belt.

  17. Real-time multiplex PCR assay for detection of Yersinia pestis and Yersinia pseudotuberculosis.

    Science.gov (United States)

    Matero, Pirjo; Pasanen, Tanja; Laukkanen, Riikka; Tissari, Päivi; Tarkka, Eveliina; Vaara, Martti; Skurnik, Mikael

    2009-01-01

    A multiplex real-time polymerase chain reaction (PCR) assay was developed for the detection of Yersinia pestis and Yersinia pseudotuberculosis. The assay includes four primer pairs, two of which are specific for Y. pestis, one for Y. pestis and Y. pseudotuberculosis and one for bacteriophage lambda; the latter was used as an internal amplification control. The Y. pestis-specific target genes in the assay were ypo2088, a gene coding for a putative methyltransferase, and the pla gene coding for the plasminogen activator. In addition, the wzz gene was used as a target to specifically identify both Y. pestis and the closely related Y. pseudotuberculosis group. The primer and probe sets described for the different genes can be used either in single or in multiplex PCR assays because the individual probes were designed with different fluorochromes. The assays were found to be both sensitive and specific; the lower limit of the detection was 10-100 fg of extracted Y. pestis or Y. pseudotuberculosis total DNA. The sensitivity of the tetraplex assay was determined to be 1 cfu for the ypo2088 and pla probe labelled with FAM and JOE fluorescent dyes, respectively.

  18. Real-Time Detection of Sporadic Meteors in the Intensified TV Imaging Systems.

    Science.gov (United States)

    Vítek, Stanislav; Nasyrova, Maria

    2017-12-29

    The automatic observation of the night sky through wide-angle video systems with the aim of detecting meteor and fireballs is currently among routine astronomical observations. The observation is usually done in multi-station or network mode, so it is possible to estimate the direction and the speed of the body flight. The high velocity of the meteorite flying through the atmosphere determines the important features of the camera systems, namely the high frame rate. Thanks to high frame rates, such imaging systems produce a large amount of data, of which only a small fragment has scientific potential. This paper focuses on methods for the real-time detection of fast moving objects in the video sequences recorded by intensified TV systems with frame rates of about 60 frames per second. The goal of our effort is to remove all unnecessary data during the daytime and make free hard-drive capacity for the next observation. The processing of data from the MAIA (Meteor Automatic Imager and Analyzer) system is demonstrated in the paper.

  19. A Finite State Machine Approach to Algorithmic Lateral Inhibition for Real-Time Motion Detection

    Directory of Open Access Journals (Sweden)

    María T. López

    2018-05-01

    Full Text Available Many researchers have explored the relationship between recurrent neural networks and finite state machines. Finite state machines constitute the best-characterized computational model, whereas artificial neural networks have become a very successful tool for modeling and problem solving. The neurally-inspired lateral inhibition method, and its application to motion detection tasks, have been successfully implemented in recent years. In this paper, control knowledge of the algorithmic lateral inhibition (ALI method is described and applied by means of finite state machines, in which the state space is constituted from the set of distinguishable cases of accumulated charge in a local memory. The article describes an ALI implementation for a motion detection task. For the implementation, we have chosen to use one of the members of the 16-nm Kintex UltraScale+ family of Xilinx FPGAs. FPGAs provide the necessary accuracy, resolution, and precision to run neural algorithms alongside current sensor technologies. The results offered in this paper demonstrate that this implementation provides accurate object tracking performance on several datasets, obtaining a high F-score value (0.86 for the most complex sequence used. Moreover, it outperforms implementations of a complete ALI algorithm and a simplified version of the ALI algorithm—named “accumulative computation”—which was run about ten years ago, now reaching real-time processing times that were simply not achievable at that time for ALI.

  20. Real-Time Hazard Detection and Avoidance Demonstration for a Planetary Lander

    Science.gov (United States)

    Epp, Chirold D.; Robertson, Edward A.; Carson, John M., III

    2014-01-01

    The Autonomous Landing Hazard Avoidance Technology (ALHAT) Project is chartered to develop and mature to a Technology Readiness Level (TRL) of six an autonomous system combining guidance, navigation and control with terrain sensing and recognition functions for crewed, cargo, and robotic planetary landing vehicles. In addition to precision landing close to a pre-mission defined landing location, the ALHAT System must be capable of autonomously identifying and avoiding surface hazards in real-time to enable a safe landing under any lighting conditions. This paper provides an overview of the recent results of the ALHAT closed loop hazard detection and avoidance flight demonstrations on the Morpheus Vertical Testbed (VTB) at the Kennedy Space Center, including results and lessons learned. This effort is also described in the context of a technology path in support of future crewed and robotic planetary exploration missions based upon the core sensing functions of the ALHAT system: Terrain Relative Navigation (TRN), Hazard Detection and Avoidance (HDA), and Hazard Relative Navigation (HRN).

  1. Detecting subsurface fluid leaks in real-time using injection and production rates

    Science.gov (United States)

    Singh, Harpreet; Huerta, Nicolas J.

    2017-12-01

    CO2 injection into geologic formations for either enhanced oil recovery or carbon storage introduces a risk for undesired fluid leakage into overlying groundwater or to the surface. Despite decades of subsurface CO2 production and injection, the technologies and methods for detecting CO2 leaks are still costly and prone to large uncertainties. This is especially true for pressure-based monitoring methods, which require the use of simplified geological and reservoir flow models to simulate the pressure behavior as well as background noise affecting pressure measurements. In this study, we propose a method to detect the time and volume of fluid leakage based on real-time measurements of well injection and production rates. The approach utilizes analogies between fluid flow and capacitance-resistance modeling. Unlike other leak detection methods (e.g. pressure-based), the proposed method does not require geological and reservoir flow models to simulate the behavior that often carry significant sources of uncertainty; therefore, with our approach the leak can be detected with greater certainty. The method can be applied to detect when a leak begins by tracking a departure in fluid production rate from the expected pattern. The method has been tuned to detect the effect of boundary conditions and fluid compressibility on leakage. To highlight the utility of this approach we use our method to detect leaks for two scenarios. The first scenario simulates a fluid leak from the storage formation into an above-zone monitoring interval. The second scenario simulates intra-reservoir migration between two compartments. We illustrate this method to detect fluid leakage in three different reservoirs with varying levels of geological and structural complexity. The proposed leakage detection method has three novelties: i) requires only readily-available data (injection and production rates), ii) accounts for fluid compressibility and boundary effects, and iii) in addition to

  2. Evaluation of four endogenous reference genes and their real-time PCR assays for common wheat quantification in GMOs detection.

    Science.gov (United States)

    Huang, Huali; Cheng, Fang; Wang, Ruoan; Zhang, Dabing; Yang, Litao

    2013-01-01

    Proper selection of endogenous reference genes and their real-time PCR assays is quite important in genetically modified organisms (GMOs) detection. To find a suitable endogenous reference gene and its real-time PCR assay for common wheat (Triticum aestivum L.) DNA content or copy number quantification, four previously reported wheat endogenous reference genes and their real-time PCR assays were comprehensively evaluated for the target gene sequence variation and their real-time PCR performance among 37 common wheat lines. Three SNPs were observed in the PKABA1 and ALMT1 genes, and these SNPs significantly decreased the efficiency of real-time PCR amplification. GeNorm analysis of the real-time PCR performance of each gene among common wheat lines showed that the Waxy-D1 assay had the lowest M values with the best stability among all tested lines. All results indicated that the Waxy-D1 gene and its real-time PCR assay were most suitable to be used as an endogenous reference gene for common wheat DNA content quantification. The validated Waxy-D1 gene assay will be useful in establishing accurate and creditable qualitative and quantitative PCR analysis of GM wheat.

  3. Evaluation of four endogenous reference genes and their real-time PCR assays for common wheat quantification in GMOs detection.

    Directory of Open Access Journals (Sweden)

    Huali Huang

    Full Text Available Proper selection of endogenous reference genes and their real-time PCR assays is quite important in genetically modified organisms (GMOs detection. To find a suitable endogenous reference gene and its real-time PCR assay for common wheat (Triticum aestivum L. DNA content or copy number quantification, four previously reported wheat endogenous reference genes and their real-time PCR assays were comprehensively evaluated for the target gene sequence variation and their real-time PCR performance among 37 common wheat lines. Three SNPs were observed in the PKABA1 and ALMT1 genes, and these SNPs significantly decreased the efficiency of real-time PCR amplification. GeNorm analysis of the real-time PCR performance of each gene among common wheat lines showed that the Waxy-D1 assay had the lowest M values with the best stability among all tested lines. All results indicated that the Waxy-D1 gene and its real-time PCR assay were most suitable to be used as an endogenous reference gene for common wheat DNA content quantification. The validated Waxy-D1 gene assay will be useful in establishing accurate and creditable qualitative and quantitative PCR analysis of GM wheat.

  4. Evaluation of Four Endogenous Reference Genes and Their Real-Time PCR Assays for Common Wheat Quantification in GMOs Detection

    Science.gov (United States)

    Huang, Huali; Cheng, Fang; Wang, Ruoan; Zhang, Dabing; Yang, Litao

    2013-01-01

    Proper selection of endogenous reference genes and their real-time PCR assays is quite important in genetically modified organisms (GMOs) detection. To find a suitable endogenous reference gene and its real-time PCR assay for common wheat (Triticum aestivum L.) DNA content or copy number quantification, four previously reported wheat endogenous reference genes and their real-time PCR assays were comprehensively evaluated for the target gene sequence variation and their real-time PCR performance among 37 common wheat lines. Three SNPs were observed in the PKABA1 and ALMT1 genes, and these SNPs significantly decreased the efficiency of real-time PCR amplification. GeNorm analysis of the real-time PCR performance of each gene among common wheat lines showed that the Waxy-D1 assay had the lowest M values with the best stability among all tested lines. All results indicated that the Waxy-D1 gene and its real-time PCR assay were most suitable to be used as an endogenous reference gene for common wheat DNA content quantification. The validated Waxy-D1 gene assay will be useful in establishing accurate and creditable qualitative and quantitative PCR analysis of GM wheat. PMID:24098735

  5. Use of quantitative real-time PCR for direct detection of serratia marcescens in marine and other aquatic environments.

    Science.gov (United States)

    Joyner, Jessica; Wanless, David; Sinigalliano, Christopher D; Lipp, Erin K

    2014-03-01

    Serratia marcescens is the etiological agent of acroporid serratiosis, a distinct form of white pox disease in the threatened coral Acropora palmata. The pathogen is commonly found in untreated human waste in the Florida Keys, which may contaminate both nearshore and offshore waters. Currently there is no direct method for detection of this bacterium in the aquatic or reef environment, and culture-based techniques may underestimate its abundance in marine waters. A quantitative real-time PCR assay was developed to detect S. marcescens directly from environmental samples, including marine water, coral mucus, sponge tissue, and wastewater. The assay targeted the luxS gene and was able to distinguish S. marcescens from other Serratia species with a reliable quantitative limit of detection of 10 cell equivalents (CE) per reaction. The method could routinely discern the presence of S. marcescens for as few as 3 CE per reaction, but it could not be reliably quantified at this level. The assay detected environmental S. marcescens in complex sewage influent samples at up to 761 CE ml(-1) and in septic system-impacted residential canals in the Florida Keys at up to 4.1 CE ml(-1). This detection assay provided rapid quantitative abilities and good sensitivity and specificity, which should offer an important tool for monitoring this ubiquitous pathogen that can potentially impact both human health and coral health.

  6. Pick- and waveform-based techniques for real-time detection of induced seismicity

    Science.gov (United States)

    Grigoli, Francesco; Scarabello, Luca; Böse, Maren; Weber, Bernd; Wiemer, Stefan; Clinton, John F.

    2018-05-01

    The monitoring of induced seismicity is a common operation in many industrial activities, such as conventional and non-conventional hydrocarbon production or mining and geothermal energy exploitation, to cite a few. During such operations, we generally collect very large and strongly noise-contaminated data sets that require robust and automated analysis procedures. Induced seismicity data sets are often characterized by sequences of multiple events with short interevent times or overlapping events; in these cases, pick-based location methods may struggle to correctly assign picks to phases and events, and errors can lead to missed detections and/or reduced location resolution and incorrect magnitudes, which can have significant consequences if real-time seismicity information are used for risk assessment frameworks. To overcome these issues, different waveform-based methods for the detection and location of microseismicity have been proposed. The main advantages of waveform-based methods is that they appear to perform better and can simultaneously detect and locate seismic events providing high-quality locations in a single step, while the main disadvantage is that they are computationally expensive. Although these methods have been applied to different induced seismicity data sets, an extensive comparison with sophisticated pick-based detection methods is still missing. In this work, we introduce our improved waveform-based detector and we compare its performance with two pick-based detectors implemented within the SeiscomP3 software suite. We test the performance of these three approaches with both synthetic and real data sets related to the induced seismicity sequence at the deep geothermal project in the vicinity of the city of St. Gallen, Switzerland.

  7. Increased efficacy for in-house validation of real-time PCR GMO detection methods.

    Science.gov (United States)

    Scholtens, I M J; Kok, E J; Hougs, L; Molenaar, B; Thissen, J T N M; van der Voet, H

    2010-03-01

    To improve the efficacy of the in-house validation of GMO detection methods (DNA isolation and real-time PCR, polymerase chain reaction), a study was performed to gain insight in the contribution of the different steps of the GMO detection method to the repeatability and in-house reproducibility. In the present study, 19 methods for (GM) soy, maize canola and potato were validated in-house of which 14 on the basis of an 8-day validation scheme using eight different samples and five on the basis of a more concise validation protocol. In this way, data was obtained with respect to the detection limit, accuracy and precision. Also, decision limits were calculated for declaring non-conformance (>0.9%) with 95% reliability. In order to estimate the contribution of the different steps in the GMO analysis to the total variation variance components were estimated using REML (residual maximum likelihood method). From these components, relative standard deviations for repeatability and reproducibility (RSD(r) and RSD(R)) were calculated. The results showed that not only the PCR reaction but also the factors 'DNA isolation' and 'PCR day' are important factors for the total variance and should therefore be included in the in-house validation. It is proposed to use a statistical model to estimate these factors from a large dataset of initial validations so that for similar GMO methods in the future, only the PCR step needs to be validated. The resulting data are discussed in the light of agreed European criteria for qualified GMO detection methods.

  8. Spectroscopic sensitivity of real-time, rapidly induced phytochemical change in response to damage.

    Science.gov (United States)

    Couture, John J; Serbin, Shawn P; Townsend, Philip A

    2013-04-01

    An ecological consequence of plant-herbivore interactions is the phytochemical induction of defenses in response to insect damage. Here, we used reflectance spectroscopy to characterize the foliar induction profile of cardenolides in Asclepias syriaca in response to damage, tracked in vivo changes and examined the influence of multiple plant traits on cardenolide concentrations. Foliar cardenolide concentrations were measured at specific time points following damage to capture their induction profile. Partial least-squares regression (PLSR) modeling was employed to calibrate cardenolide concentrations to reflectance spectroscopy. In addition, subsets of plants were either repeatedly sampled to track in vivo changes or modified to reduce latex flow to damaged areas. Cardenolide concentrations and the induction profile of A. syriaca were well predicted using models derived from reflectance spectroscopy, and this held true for repeatedly sampled plants. Correlations between cardenolides and other foliar-related variables were weak or not significant. Plant modification for latex reduction inhibited an induced cardenolide response. Our findings show that reflectance spectroscopy can characterize rapid phytochemical changes in vivo. We used reflectance spectroscopy to identify the mechanisms behind the production of plant secondary metabolites, simultaneously characterizing multiple foliar constituents. In this case, cardenolide induction appears to be largely driven by enhanced latex delivery to leaves following damage. © 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.

  9. Rapid testing and identification of actuator using dSPACE real-time emulator

    Science.gov (United States)

    Xie, Daocheng; Wang, Zhongwei; Zeng, Qinghua

    2011-10-01

    To solve the problem of model identification of actuator in control system design of aerocraft, testing system based on dSPACE emulator is established, sending testing signal and receiving feedback voltage are realized using dSPACE interactive cards, communication between signal generating equipment and feedback voltage acquisition equipment is synchronized. This paper introduces the hardware architecture and key technologies of the simulation system. Constructing, downloading and calculating of the testing model is finished using dSPACE emulator, D/A transfer of testing signal is realized using DS2103 card, DS2002 card transfer the feedback voltage to digital value. Filtering module is added to the signal acquisition, for reduction of noise interference in the A/D channel. Precision of time and voltage is improved by setting acquisition period 1ms. The data gathered is recorded and displayed with Controldesk tools. The response of four actuators under different frequency are tested, frequency-domain analysis is done using least square method, the model of actuator is identified, simulation data fits well with real response of the actuator. The testing system created with dSPACE emulator satisfies the rapid testing and identification of actuator.

  10. Comparison of Hybrid Capture 2 Assay with Real-time-PCR for Detection and Quantitation of Hepatitis B Virus DNA.

    Science.gov (United States)

    Majid, Farjana; Jahan, Munira; Lutful Moben, Ahmed; Tabassum, Shahina

    2014-01-01

    Both real-time-polymerase chain reaction (PCR) and hybrid capture 2 (HC2) assay can detect and quantify hepatitis B virus (HBV) DNA. However, real-time-PCR can detect a wide range of HBV DNA, while HC2 assay could not detect lower levels of viremia. The present study was designed to detect and quantify HBV DNA by real-time-PCR and HC2 assay and compare the quantitative data of these two assays. A cross-sectional study was conducted in between July 2010 and June 2011. A total of 66 serologically diagnosed chronic hepatitis B (CHB) patients were selected for the study. Real-time-PCR and HC2 assay was done to detect HBV DNA. Data were analyzed by statistical Package for the social sciences (SPSS). Among 66 serologically diagnosed chronic hepatitis B patients 40 (60.61%) patients had detectable and 26 (39.39%) had undetectable HBV DNA by HC2 assay. Concordant results were obtained for 40 (60.61%) out of these 66 patients by real-time-PCR and HC2 assay with mean viral load of 7.06 ± 1.13 log 10 copies/ml and 6.95 ± 1.08 log 10 copies/ml, respectively. In the remaining 26 patients, HBV DNA was detectable by real-time-PCR in 20 patients (mean HBV DNA level was 3.67 ± 0.72 log 10 copies/ml. However, HBV DNA could not be detectable in six cases by the both assays. The study showed strong correlation (r = 0.915) between real-time-PCR and HC2 assay for the detection and quantification of HBV DNA. HC2 assay may be used as an alternative to real-time-PCR for CHB patients. How to cite this article: Majid F, Jahan M, Moben AL, Tabassum S. Comparison of Hybrid Capture 2 Assay with Real-time-PCR for Detection and Quantitation of Hepatitis B Virus DNA. Euroasian J Hepato-Gastroenterol 2014;4(1):31-35.

  11. Gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of Japanese encephalitis virus

    International Nuclear Information System (INIS)

    Huang, S-H; Tsai, M-H; Lin, C-W; Yang, T-C; Chuang, P-H; Tsai, I-S; Lu, H-C; Wan Lei; Lin, Y-J; Lai, C-H

    2008-01-01

    Virus isolation and antibody detection are routinely used for diagnosis of Japanese encephalitis virus (JEV) infection, but the low level of transient viremia in some JE patients makes JEV isolation from clinical and surveillance samples very difficult. We describe the use of gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of JEV from its RNA genome. We tested the effect of gold nanoparticles on four different PCR systems, including conventional PCR, reverse-transcription PCR (RT-PCR), and SYBR green real-time PCR and RT-PCR assays for diagnosis in the acute phase of JEV infection. Gold nanoparticles increased the amplification yield of the PCR product and shortened the PCR time compared to the conventional reaction. In addition, nanogold-based real-time RT-PCR showed a linear relationship between Ct and template amount using ten-fold dilutions of JEV. The nanogold-based RT-PCR and real-time quantitative RT-PCR assays were able to detect low levels (1-10 000 copies) of the JEV RNA genomes extracted from culture medium or whole blood, providing early diagnostic tools for the detection of low-level viremia in the acute-phase infection. The assays described here were simple, sensitive, and rapid approaches for detection and quantitation of JEV in tissue cultured samples as well as clinical samples

  12. Real-Time Digital Bright Field Technology for Rapid Antibiotic Susceptibility Testing.

    Science.gov (United States)

    Canali, Chiara; Spillum, Erik; Valvik, Martin; Agersnap, Niels; Olesen, Tom

    2018-01-01

    Optical scanning through bacterial samples and image-based analysis may provide a robust method for bacterial identification, fast estimation of growth rates and their modulation due to the presence of antimicrobial agents. Here, we describe an automated digital, time-lapse, bright field imaging system (oCelloScope, BioSense Solutions ApS, Farum, Denmark) for rapid and higher throughput antibiotic susceptibility testing (AST) of up to 96 bacteria-antibiotic combinations at a time. The imaging system consists of a digital camera, an illumination unit and a lens where the optical axis is tilted 6.25° relative to the horizontal plane of the stage. Such tilting grants more freedom of operation at both high and low concentrations of microorganisms. When considering a bacterial suspension in a microwell, the oCelloScope acquires a sequence of 6.25°-tilted images to form an image Z-stack. The stack contains the best-focus image, as well as the adjacent out-of-focus images (which contain progressively more out-of-focus bacteria, the further the distance from the best-focus position). The acquisition process is repeated over time, so that the time-lapse sequence of best-focus images is used to generate a video. The setting of the experiment, image analysis and generation of time-lapse videos can be performed through a dedicated software (UniExplorer, BioSense Solutions ApS). The acquired images can be processed for online and offline quantification of several morphological parameters, microbial growth, and inhibition over time.

  13. Wireless and real-time structural damage detection: A novel decentralized method for wireless sensor networks

    Science.gov (United States)

    Avci, Onur; Abdeljaber, Osama; Kiranyaz, Serkan; Hussein, Mohammed; Inman, Daniel J.

    2018-06-01

    Being an alternative to conventional wired sensors, wireless sensor networks (WSNs) are extensively used in Structural Health Monitoring (SHM) applications. Most of the Structural Damage Detection (SDD) approaches available in the SHM literature are centralized as they require transferring data from all sensors within the network to a single processing unit to evaluate the structural condition. These methods are found predominantly feasible for wired SHM systems; however, transmission and synchronization of huge data sets in WSNs has been found to be arduous. As such, the application of centralized methods with WSNs has been a challenge for engineers. In this paper, the authors are presenting a novel application of 1D Convolutional Neural Networks (1D CNNs) on WSNs for SDD purposes. The SDD is successfully performed completely wireless and real-time under ambient conditions. As a result of this, a decentralized damage detection method suitable for wireless SHM systems is proposed. The proposed method is based on 1D CNNs and it involves training an individual 1D CNN for each wireless sensor in the network in a format where each CNN is assigned to process the locally-available data only, eliminating the need for data transmission and synchronization. The proposed damage detection method operates directly on the raw ambient vibration condition signals without any filtering or preprocessing. Moreover, the proposed approach requires minimal computational time and power since 1D CNNs merge both feature extraction and classification tasks into a single learning block. This ability is prevailingly cost-effective and evidently practical in WSNs considering the hardware systems have been occasionally reported to suffer from limited power supply in these networks. To display the capability and verify the success of the proposed method, large-scale experiments conducted on a laboratory structure equipped with a state-of-the-art WSN are reported.

  14. Event Detection Intelligent Camera: Demonstration of flexible, real-time data taking and processing

    Energy Technology Data Exchange (ETDEWEB)

    Szabolics, Tamás, E-mail: szabolics.tamas@wigner.mta.hu; Cseh, Gábor; Kocsis, Gábor; Szepesi, Tamás; Zoletnik, Sándor

    2015-10-15

    Highlights: • We present EDICAM's operation principles description. • Firmware tests results. • Software test results. • Further developments. - Abstract: An innovative fast camera (EDICAM – Event Detection Intelligent CAMera) was developed by MTA Wigner RCP in the last few years. This new concept was designed for intelligent event driven processing to be able to detect predefined events and track objects in the plasma. The camera provides a moderate frame rate of 400 Hz at full frame resolution (1280 × 1024), and readout of smaller region of interests can be done in the 1–140 kHz range even during exposure of the full image. One of the most important advantages of this hardware is a 10 Gbit/s optical link which ensures very fast communication and data transfer between the PC and the camera, enabling two level of processing: primitive algorithms in the camera hardware and high-level processing in the PC. This camera hardware has successfully proven to be able to monitoring the plasma in several fusion devices for example at ASDEX Upgrade, KSTAR and COMPASS with the first version of firmware. A new firmware and software package is under development. It allows to detect predefined events in real time and therefore the camera is capable to change its own operation or to give warnings e.g. to the safety system of the experiment. The EDICAM system can handle a huge amount of data (up to TBs) with high data rate (950 MB/s) and will be used as the central element of the 10 camera overview video diagnostic system of Wendenstein 7-X (W7-X) stellarator. This paper presents key elements of the newly developed built-in intelligence stressing the revolutionary new features and the results of the test of the different software elements.

  15. Real-time person detection in low-resolution thermal infrared imagery with MSER and CNNs

    Science.gov (United States)

    Herrmann, Christian; Müller, Thomas; Willersinn, Dieter; Beyerer, Jürgen

    2016-10-01

    LWIR imagery applications and is capable of fast classification. Evaluation on several different LWIR person detection datasets shows an error rate reduction of up to 80 percent compared to previous approaches consisting of MSER, local image descriptors and a standard classifier such as an SVM or boosted decision trees. Further time measurements show that the proposed processing chain is capable of real-time person detection in LWIR camera streams.

  16. Real-time porphyrin detection in plaque and caries: a case study

    Science.gov (United States)

    Timoshchuk, Mari-Alina I.; Ridge, Jeremy S.; Rugg, Amanda L.; Nelson, Leonard Y.; Kim, Amy S.; Seibel, Eric J.

    2015-02-01

    An ultrathin scanning fiber endoscope, originally developed for cancer diagnosis, was used in a case study to locate plaque and caries. The imaging system incorporated software mitigation of background auto-fluorescence (AF). In conventional fluorescence imaging, varying AF across a tooth surface can mask low-level porphyrin signals. Laser-induced auto-fluorescence signals of dental tissue excited using a 405-nm laser typically produce fluorescence over a wavelength range extending from 440-nm to 750-nm. Anaerobic bacterial metabolism produces various porphyrin species (eg. protoporphyrin IX) that are located in carious enamel, dentin, gingivitis sites, and plaque. In our case study, these porphyrin deposits remained as long as one day after prophylaxis. Imaging the tooth surface using 405-nm excitation and subtracting the natural AF enhances the image contrast of low-level porphyrin deposits, which would otherwise be masked by the high background AF. In a case study, healthy tissues as well as sites of early and advanced caries formations were scanned for visual and quantitative signs of red fluorescence associated with porphyrin species using a background mitigation algorithm. Initial findings show increasing amplitudes of red fluorescence as caries severity increases from early to late stages. Sites of plaque accumulation also displayed red fluorescence similar to that found in carious dental tissue. The use of real-time background mitigation of natural dental AF can enhance the detection of low porphyrin concentrations that are indicators of early stage caries formation.

  17. Real-time in vivo detection of biomaterial-induced reactive oxygen species.

    Science.gov (United States)

    Liu, Wendy F; Ma, Minglin; Bratlie, Kaitlin M; Dang, Tram T; Langer, Robert; Anderson, Daniel G

    2011-03-01

    The non-specific host response to implanted biomaterials is often a key challenge of medical device design. To evaluate biocompatibility, measuring the release of reactive oxygen species (ROS) produced by inflammatory cells in response to biomaterial surfaces is a well-established method. However, the detection of ROS in response to materials implanted in vivo has not yet been demonstrated. Here, we develop a bioluminescence whole animal imaging approach to observe ROS released in response to subcutaneously-implanted materials in live animals. We compared the real-time generation of ROS in response to two representative materials, polystyrene and alginate, over the course of 28 days. High levels of ROS were observed near polystyrene, but not alginate implants, and persisted throughout the course of 28 days. Histological analysis revealed that high levels of ROS correlated not only with the presence of phagocytic cells at early timepoints, but also fibrosis at later timepoints, suggesting that ROS may be involved in both the acute and chronic phase of the foreign body response. These data are the first in vivo demonstration of ROS generation in response to implanted materials, and describe a novel technique to evaluate the host response. Copyright © 2010 Elsevier Ltd. All rights reserved.

  18. Real-time Physiological Emotion Detection Mechanisms: Effects of Exercise and Affect Intensity.

    Science.gov (United States)

    Leon, E; Clarke, G; Sepulveda, F; Callaghan, V

    2005-01-01

    The development of systems capable of recognizing and categorising emotions is of interest to researchers in various scientific areas including artificial intelligence. The traditional notion that emotions and rationality are two separate realms has gradually been challenged. The work of neurologists has shown the strong relationship between emotional episodes and the way humans think and act. Furthermore, emotions not only regulate human decisions but could also contribute to a more satisfactory response to the environment, i.e., faster and more precise actions. In this paper an analysis of physiological signals employed in real-time emotion detection is presented in the context of Intelligent Inhabited Environments (IIE). Two studies were performed to investigate whether physical exertion has a significant effect on bodily signals stemming from emotional episodes with subjects having various degrees of affect intensity: 1) a statistical analysis using the Wilcoxon Test, and 2) a cluster analysis using the Davies-Bouldin Index. Preliminary results demonstrated that the heart rate and skin resistance consistently showed similar changes regardless of the physical stimuli while blood volume pressure did not show a significant change. It was also found that neither physical stress nor affect intensity played a role in the separation of neutral and non-neutral emotional states.

  19. Apple detection using infrared thermal image, 3: Real-time temperature measurement of apple tree

    International Nuclear Information System (INIS)

    Zhang, S.H.; Takahashi, T.; Fukuchi, H.; Sun, M.; Terao, H.

    1998-01-01

    In Part 1, we reported the thermal distribution characteristics and the identification methods of apples, leaves and branches by using the infrared thermal image at the specific time. This paper reports the temperature changing characteristics and the relationships among apples, leaves and air temperature based on the information measured by the infrared thermal image equipment in the real-time for 24 hours. As a result, it was confirmed that the average temperature of apples was 1 degree C or more higher than the one of the leaves, and the average temperature of the leaves was almost same as the air temperature within daytime and about 3 hours period after sunset. It was also clarified for a remarkable temperature difference not to exist for midnight and the early morning between the apples and the leaves, and both became almost as well as the air temperature. Moreover, a binary image was easily obtained and the apples could be detected by using this temperature difference informat

  20. Real-time camera-based face detection using a modified LAMSTAR neural network system

    Science.gov (United States)

    Girado, Javier I.; Sandin, Daniel J.; DeFanti, Thomas A.; Wolf, Laura K.

    2003-03-01

    This paper describes a cost-effective, real-time (640x480 at 30Hz) upright frontal face detector as part of an ongoing project to develop a video-based, tetherless 3D head position and orientation tracking system. The work is specifically targeted for auto-stereoscopic displays and projection-based virtual reality systems. The proposed face detector is based on a modified LAMSTAR neural network system. At the input stage, after achieving image normalization and equalization, a sub-window analyzes facial features using a neural network. The sub-window is segmented, and each part is fed to a neural network layer consisting of a Kohonen Self-Organizing Map (SOM). The output of the SOM neural networks are interconnected and related by correlation-links, and can hence determine the presence of a face with enough redundancy to provide a high detection rate. To avoid tracking multiple faces simultaneously, the system is initially trained to track only the face centered in a box superimposed on the display. The system is also rotationally and size invariant to a certain degree.

  1. Deciding between compensated volume balance and real time transient models for pipeline leak detection system

    Energy Technology Data Exchange (ETDEWEB)

    Baptista, Renan Martins [PETROBRAS, Rio de Janeiro, RJ (Brazil). Centro de Pesquisas. Div. de Explotacao]. E-mail: renan@cenpes.petrobras.com.br

    2000-07-01

    This paper describes a technical procedure to assess a software based leak detection system (LDS), by deciding between a simpler low cost, less effective product, having a fast installation and tuning, and a complex one with high cost and efficiency, which however takes a long time to be properly installed. This is a common decision among the pipeline operating companies, considering that the majority of the lines are short, with single phase liquid flow (which may include batches), basic communication system and instrumentation. Service companies offer realistic solutions for liquid flow, but usually designed to big pipeline networks, flowing multiple batches and allowing multiple fluid entrances and deliveries. Those solutions are sometimes impractical to short pipelines, due to its high cost, as well as long tuning procedures, complex instrumentation, communication and computer requirements. It is intended to approach here the best solution according to its cost. In a practical sense, it means to differentiate the various LDS techniques. Those techniques are available in a considerable number, and they are still spreading, according to the different scenarios. However, two most known and worldwide implemented techniques hold the majority of the market: the Compensated Volume Balance (CVB), which is less accurate, reliable and robust, but cheaper, simpler and faster to install, and the Real Time Transient Model (RTTM), which is very reliable, accurate and robust, but expensive and complex. This work will describe a way to define whether one can use or not a CVB in a pipeline. (author)

  2. Fast Real-Time PCR for the Detection of Crustacean Allergens in Foods.

    Science.gov (United States)

    Santaclara, Francisco J; Espiñeira, Montserrat

    2017-01-01

    Crustaceans are one of the most common allergens causing severe food reaction. Hypersensitivity reactions associated with seafood intake are one of the most common food allergies in adults. Crustaceans including shrimps, prawns, crabs, lobster, and crayfish are a common cause of anaphylaxis or hypersensitivity, with shrimps and crabs being the most common causes of allergy. Symptoms occur most often when food or cooking water are ingested.These food allergens are a health problem, and they have become very important; as evidenced by the existence of several regulations that establish that labeling must be present regarding these allergens to warn consumers.The methodology herein exposed allows the detection of crustaceans in any type of product, including those where very aggressive treatments of temperature and pressure are used during the manufacturing process.The main features of this method are its high sensitivity and specificity, and reduced analysis time of real-time PCR (40 min). This assay is a potential tool in issues related to the labeling of products and food security to protect the allergic consumer.

  3. Detection and Characterization of Equatorial Scintillation for Real-Time Operational Support

    National Research Council Canada - National Science Library

    McNeil, W

    1997-01-01

    The Phillips Laboratory Scintillation Network Decision Aid (PL-SCINDA) is a software tool which uses real-time data from remote sites to model ionospheric plasma depletions in the equatorial region...

  4. Detection and typing of highly pathogenic porcine reproductive and respiratory syndrome virus by multiplex real-time rt-PCR.

    Directory of Open Access Journals (Sweden)

    Kerstin Wernike

    Full Text Available Porcine reproductive and respiratory syndrome (PRRS causes economic losses in the pig industry worldwide, and PRRS viruses (PRRSV are classified into the two distinct genotypes "North American (NA, type 2" and "European (EU, type 1". In 2006, a highly pathogenic NA strain of PRRSV (HP-PRRSV, characterized by high fever as well as high morbidity and mortality, emerged in swine farms in China. Therefore, a real-time reverse transcription polymerase chain reaction (RT-qPCR assay specific for HP-PRRSV was developed and combined with type 1- and type 2-specific RT-qPCR systems. Furthermore, an internal control, based on a heterologous RNA, was successfully introduced. This final multiplex PRRSV RT-qPCR, detecting and typing PRRSV, had an analytical sensitivity of less than 200 copies per µl for the type 1-assay and 20 copies per µl for the type 2- and HP assays and a high diagnostic sensitivity. A panel of reference strains and field isolates was reliably detected and samples from an animal trial with a Chinese HP-PRRS strain were used for test validation. The new multiplex PRRSV RT-qPCR system allows for the first time the highly sensitive detection and rapid differentiation of PRRSV of both genotypes as well as the direct detection of HP-PRRSV.

  5. Development of a real-time PCR assay for detection of planktonic red king crab (Paralithodes camtschaticus (Tilesius 1815)) larvae

    Science.gov (United States)

    Jensen, Pamela C.; Purcell, Maureen K.; Morado, J. Frank; Eckert, Ginny L.

    2012-01-01

    The Alaskan red king crab (Paralithodes camtschaticus) fishery was once one of the most economically important single-species fisheries in the world, but is currently depressed. This fishery would benefit from improved stock assessment capabilities. Larval crab distribution is patchy temporally and spatially, requiring extensive sampling efforts to locate and track larval dispersal. Large-scale plankton surveys are generally cost prohibitive because of the effort required for collection and the time and taxonomic expertise required to sort samples to identify plankton individually via light microscopy. Here, we report the development of primers and a dual-labeled probe for use in a DNA-based real-time polymerase chain reaction assay targeting the red king crab, mitochondrial gene cytochrome oxidase I for the detection of red king crab larvae DNA in plankton samples. The assay allows identification of plankton samples containing crab larvae DNA and provides an estimate of DNA copy number present in a sample without sorting the plankton sample visually. The assay was tested on DNA extracted from whole red king crab larvae and plankton samples seeded with whole larvae, and it detected DNA copies equivalent to 1/10,000th of a larva and 1 crab larva/5mL sieved plankton, respectively. The real-time polymerase chain reaction assay can be used to screen plankton samples for larvae in a fraction of the time required for traditional microscopial methods, which offers advantages for stock assessment methodologies for red king crab as well as a rapid and reliable method to assess abundance of red king crab larvae as needed to improve the understanding of life history and population processes, including larval population dynamics.

  6. Impacts of Earth rotation parameters on GNSS ultra-rapid orbit prediction: Derivation and real-time correction

    Science.gov (United States)

    Wang, Qianxin; Hu, Chao; Xu, Tianhe; Chang, Guobin; Hernández Moraleda, Alberto

    2017-12-01

    Analysis centers (ACs) for global navigation satellite systems (GNSSs) cannot accurately obtain real-time Earth rotation parameters (ERPs). Thus, the prediction of ultra-rapid orbits in the international terrestrial reference system (ITRS) has to utilize the predicted ERPs issued by the International Earth Rotation and Reference Systems Service (IERS) or the International GNSS Service (IGS). In this study, the accuracy of ERPs predicted by IERS and IGS is analyzed. The error of the ERPs predicted for one day can reach 0.15 mas and 0.053 ms in polar motion and UT1-UTC direction, respectively. Then, the impact of ERP errors on ultra-rapid orbit prediction by GNSS is studied. The methods for orbit integration and frame transformation in orbit prediction with introduced ERP errors dominate the accuracy of the predicted orbit. Experimental results show that the transformation from the geocentric celestial references system (GCRS) to ITRS exerts the strongest effect on the accuracy of the predicted ultra-rapid orbit. To obtain the most accurate predicted ultra-rapid orbit, a corresponding real-time orbit correction method is developed. First, orbits without ERP-related errors are predicted on the basis of ITRS observed part of ultra-rapid orbit for use as reference. Then, the corresponding predicted orbit is transformed from GCRS to ITRS to adjust for the predicted ERPs. Finally, the corrected ERPs with error slopes are re-introduced to correct the predicted orbit in ITRS. To validate the proposed method, three experimental schemes are designed: function extrapolation, simulation experiments, and experiments with predicted ultra-rapid orbits and international GNSS Monitoring and Assessment System (iGMAS) products. Experimental results show that using the proposed correction method with IERS products considerably improved the accuracy of ultra-rapid orbit prediction (except the geosynchronous BeiDou orbits). The accuracy of orbit prediction is enhanced by at least 50

  7. Evaluation of a rapid, real-time intrapartum group B Streptococcus assay

    Science.gov (United States)

    YOUNG, Brett C.; DODGE, Laura E.; GUPTA, Munish; RHEE, Julie S.; HACKER, Michele R.

    2011-01-01

    OBJECTIVE To evaluate an intrapartum nucleic acid amplification test (NAAT) for Group B streptococcus (GBS). STUDY DESIGN Prospective cohort study of 559 women comparing intrapartum GBS culture with antepartum culture and intrapartum NAAT. RESULTS GBS prevalence was 19.5% by antepartum culture and 23.8% by intrapartum culture. Compared with intrapartum culture, antepartum culture had 69.2% sensitivity (60.6–76.9) and 96.0% specificity (93.7–97.7). The NAAT demonstrated sensitivity of 90.8% (84.6–95.2), specificity of 97.6% (95.6–98.8), and predictive values exceeding 92%. The incidence of discordant cultures was 10.4%. Of the women with negative antepartum and positive intrapartum cultures, only 1 (2.4%) received intrapartum antibiotics. Compared with white women, black (P=0.02) and Hispanic (P=0.02) women were more likely to have discordant cultures. CONCLUSION This intrapartum NAAT has excellent characteristics. It may be superior to antepartum culture for detecting intrapartum GBS—allowing more accurate management of laboring mothers and reducing neonatal GBS sepsis. PMID:21864820

  8. Automated real time peg and tool detection for the FLS trainer box.

    Science.gov (United States)

    Nemani, Arun; Sankaranarayanan, Ganesh

    2012-01-01

    This study proposes a method that effectively tracks trocar tool and peg positions in real time to allow real time assessment of the peg transfer task of the Fundamentals of Laparoscopic Surgery (FLS). By utilizing custom code along with OpenCV libraries, tool and peg positions can be accurately tracked without altering the original setup conditions of the FLS trainer box. This is achieved via a series of image filtration sequences, thresholding functions, and Haar training methods.

  9. Attention focussing and anomaly detection in real-time systems monitoring

    Science.gov (United States)

    Doyle, Richard J.; Chien, Steve A.; Fayyad, Usama M.; Porta, Harry J.

    1993-01-01

    In real-time monitoring situations, more information is not necessarily better. When faced with complex emergency situations, operators can experience information overload and a compromising of their ability to react quickly and correctly. We describe an approach to focusing operator attention in real-time systems monitoring based on a set of empirical and model-based measures for determining the relative importance of sensor data.

  10. Integrated Oil spill detection and forecasting using MOON real time data

    OpenAIRE

    De Dominicis, M.; Pinardi, N.; Coppini, G.; Tonani, M.; Guarnieri, A.; Zodiatis, G.; Lardner, R.; Santoleri, R.

    2009-01-01

    MOON (Mediterranean Operational Oceanography Network) is an operational distributed system ready to provide quality controlled and timely marine observations (in situ and satellite) and environmental analyses and predictions for management of oil spill accidents. MOON operational systems are based upon the real time functioning of an integrated system composed of the Real Time Observing system, the regional, sub-regional and coastal forecasting systems and a products dissemination system. All...

  11. Development of duplex real-time RT-PCR based on Taqman technology for detecting simultaneously the genome of pan-enterovirus and enterovirus 71.

    Science.gov (United States)

    Hwang, Seoyeon; Kang, Byunghak; Hong, Jiyoung; Kim, Ahyoun; Kim, Hyejin; Kim, Kisang; Cheon, Doo-Sung

    2013-07-01

    Human enterovirus (EV) 71 is the main etiological agent of hand, foot, and mouth disease (HFMD). It is associated with neurological complications, and caused fatalities during recent outbreaks in the Asia-Pacific region. Infections caused by EV71 could lead to many complications, ranging from brainstem encephalitis to pulmonary oedema, resulting in high mortality. In this study, a duplex real-time RT-PCR assay was developed in order to simultaneously detect pan-EV and EV71. EV71-specific primers and probes were designed based on the highly conserved VP1 region of EV71. Five EV71 strains were detected as positive, and no positive fluorescence signal was observed in the duplex real-time RT-PCR for other viral RNA, which showed 100% specificity for the selected panel, and no cross-reactions were observed in this duplex real-time RT-PCR. The EV71-specific duplex real-time RT-PCR was more sensitive than conventional RT-PCR, and detected viral titers that were 10-fold lower than those measured by the latter. Of the 381 HFMD clinical specimens, 196 (51.4%) cases were pan-EV-positive, of which 170 (86.7%) were EV71-positive when tested by pan-EV and EV71-specific duplex real-time RT-PCR. EV71-specific duplex real-time RT-PCR offers a rapid and sensitive method to detect EV71 from clinical specimens, and will allow quarantine measures to be taken more effectively during outbreaks. Copyright © 2013 Wiley Periodicals, Inc.

  12. Development of a high-speed real-time PCR system for rapid and precise nucleotide recognition

    Science.gov (United States)

    Terazono, Hideyuki; Takei, Hiroyuki; Hattori, Akihiro; Yasuda, Kenji

    2010-04-01

    Polymerase chain reaction (PCR) is a common method used to create copies of a specific target region of a DNA sequence and to produce large quantities of DNA. A few DNA molecules, which act as templates, are rapidly amplified by PCR into many billions of copies. PCR is a key technology in genome-based biological analysis, revolutionizing many life science fields such as medical diagnostics, food safety monitoring, and countermeasures against bioterrorism. Thus, many applications have been developed with the thermal cycling. For these PCR applications, one of the most important key factors is reduction in the data acquisition time. To reduce the acquisition time, it is necessary to decrease the temperature transition time between the high and low ends as much as possible. We have developed a novel rapid real-time PCR system based on rapid exchange of media maintained at different temperatures. This system consists of two thermal reservoirs and a reaction chamber for PCR observation. The temperature transition was achieved within 0.3 sec, and good thermal stability was achieved during thermal cycling with rapid exchange of circulating media. This system allows rigorous optimization of the temperatures required for each stage of the PCR processes. Resulting amplicons were confirmed by electrophoresis. Using the system, rapid DNA amplification was accomplished within 3.5 min, including initial heating and complete 50 PCR cycles. It clearly shows that the device could allow us faster temperature switching than the conventional conduction-based heating systems based on Peltier heating/cooling.

  13. Real time PCR to detect the environmental faecal contamination by Echinococcus multilocularis from red fox stools.

    Science.gov (United States)

    Knapp, Jenny; Millon, Laurence; Mouzon, Lorane; Umhang, Gérald; Raoul, Francis; Ali, Zeinaba Said; Combes, Benoît; Comte, Sébastien; Gbaguidi-Haore, Houssein; Grenouillet, Frédéric; Giraudoux, Patrick

    2014-03-17

    The oncosphere stage of Echinococcus multilocularis in red fox stools can lead, after ingestion, to the development of alveolar echinococcosis in the intermediate hosts, commonly small mammals and occasionally humans. Monitoring animal infection and environmental contamination is a key issue in public health surveillance. We developed a quantitative real-time PCR technique (qPCR) to detect and quantify E. multilocularis DNA released in fox faeces. A qPCR technique using a hydrolysis probe targeting part of the mitochondrial gene rrnL was assessed on (i) a reference collection of stools from 57 necropsied foxes simultaneously investigated using the segmental sedimentation and counting technique (SSCT) (29 positive for E. multilocularis worms and 28 negative animals for the parasite); (ii) a collection of 114 fox stools sampled in the field: two sets of 50 samples from contrasted endemic regions in France and 14 from an E. multilocularis-free area (Greenland). Of the negative SSCT controls, 26/28 were qPCR-negative and two were weakly positive. Of the positive SSCT foxes, 25/29 samples were found to be positive by qPCR. Of the field samples, qPCR was positive in 21/50 (42%) and 5/48 (10.4%) stools (2 samples inhibited), originating respectively from high and low endemic areas. In faeces, averages of 0.1 pg/μl of DNA in the Jura area and 0.7 pg/μl in the Saône-et-Loire area were detected. All qPCR-positive samples were confirmed by sequencing. The qPCR technique developed here allowed us to quantify environmental E. multilocularis contamination by fox faeces by studying the infectious agent directly. No previous study had performed this test in a one-step reaction. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Telemetric real-time sensor for the detection of acute upper gastrointestinal bleeding.

    Science.gov (United States)

    Schostek, Sebastian; Zimmermann, Melanie; Keller, Jan; Fode, Mario; Melbert, Michael; Schurr, Marc O; Gottwald, Thomas; Prosst, Ruediger L

    2016-04-15

    Acute upper gastrointestinal bleedings from ulcers or esophago-gastric varices are life threatening medical conditions which require immediate endoscopic therapy. Despite successful endoscopic hemostasis, there is a significant risk of rebleeding often requiring close surveillance of these patients in the intensive care unit (ICU). Any time delay to recognize bleeding may lead to a high blood loss and increases the risk of death. A novel telemetric real-time bleeding sensor can help indicate blood in the stomach: the sensor is swallowed to detect active bleeding or is anchored endoscopically on the gastrointestinal wall close to the potential bleeding source. By telemetric communication with an extra-corporeal receiver, information about the bleeding status is displayed. In this study the novel sensor, which measures characteristic optical properties of blood, has been evaluated in an ex-vivo setting to assess its clinical applicability and usability. Human venous blood of different concentrations, various fluids, and liquid food were tested. The LED-based sensor was able to reliably distinguish between concentrated blood and other liquids, especially red-colored fluids. In addition, the spectrometric quality of the small sensor (size: 6.5mm in diameter, 25.5mm in length) was comparable to a much larger and technically more complex laboratory spectrophotometer. The experimental data confirm the capability of a miniaturized sensor to identify concentrated blood, which could help in the very near future the detection of upper gastrointestinal bleeding and to survey high-risk patients for rebleeding. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Application of a molecular beacon based real-time isothermal amplification (MBRTIA) technology for simultaneous detection of Bacillus cereus and Staphylococcus aureus.

    Science.gov (United States)

    Mandappa, I M; Joglekar, Prasanna; Manonmani, H K

    2015-07-01

    A multiplex real-time isothermal amplification assay was developed using molecular beacons for the detection of Bacillus cereus and Staphylococcus aureus by targeting four important virulence genes. A correlation between targeting highly accessible DNA sequences and isothermal amplification based molecular beacon efficiency and sensitivity was demonstrated using phi(Φ)29 DNA polymerase at a constant isothermal temperature of 30 °C. It was very selective and consistently detected down to 10(1) copies of DNA. The specificity and sensitivity of this assay, when tested with pure culture were high, surpassing those of currently used PCR assays for the detection of these organisms. The molecular beacon based real-time isothermal amplification (MBRTIA) assay could be carried out entirely in 96 well plates or well strips, enabling a rapid and high-throughput detection of food borne pathogens.

  16. Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor

    DEFF Research Database (Denmark)

    Jensen, Pia Wrensted; Falconi, Mattia; Kristoffersen, Emil Laust

    2013-01-01

    structure of the biosensor. The specific action of TDP1 removes the quencher, thereby enabling optical detection of the fluorophore. Since the enzymatic action of TDP1 is the only “signal amplification” the increase in fluorescence may easily be followed in real-time and allows quantitative analyses of TDP1......Real-time detection of enzyme activities may present the easiest and most reliable way of obtaining quantitative analyses in biological samples. We present a new DNA-biosensor capable of detecting the activity of the potential anticancer drug target tyrosyl-DNA phosphodiesterase 1 (TDP1) in a very...... simple, high throughput, and real-time format. The biosensor is specific for Tdp1 even in complex biological samples, such as human cell extracts, and may consequently find future use in fundamental studies as well as a cancer predictive tool allowing fast analyses of diagnostic cell samples...

  17. Development of novel algorithm and real-time monitoring ambulatory system using Bluetooth module for fall detection in the elderly.

    Science.gov (United States)

    Hwang, J Y; Kang, J M; Jang, Y W; Kim, H

    2004-01-01

    Novel algorithm and real-time ambulatory monitoring system for fall detection in elderly people is described. Our system is comprised of accelerometer, tilt sensor and gyroscope. For real-time monitoring, we used Bluetooth. Accelerometer measures kinetic force, tilt sensor and gyroscope estimates body posture. Also, we suggested algorithm using signals which obtained from the system attached to the chest for fall detection. To evaluate our system and algorithm, we experimented on three people aged over 26 years. The experiment of four cases such as forward fall, backward fall, side fall and sit-stand was repeated ten times and the experiment in daily life activity was performed one time to each subject. These experiments showed that our system and algorithm could distinguish between falling and daily life activity. Moreover, the accuracy of fall detection is 96.7%. Our system is especially adapted for long-time and real-time ambulatory monitoring of elderly people in emergency situation.

  18. A triplex quantitative real-time PCR assay for differential detection of human adenovirus serotypes 2, 3 and 7.

    Science.gov (United States)

    Qiu, Fang-Zhou; Shen, Xin-Xin; Zhao, Meng-Chuan; Zhao, Li; Duan, Su-Xia; Chen, Chen; Qi, Ju-Ju; Li, Gui-Xia; Wang, Le; Feng, Zhi-Shan; Ma, Xue-Jun

    2018-05-02

    Human adenovirus (HAdV) serotypes 2, 3 and 7 are more prevalent than other serotypes and have been associated with severe pneumonia in pediatric children. Molecular typing of HAdV is not routinely performed in clinical diagnostic laboratories as it is time-consuming and labor-intensive. In the present study, we developed a triplex quantitative real-time PCR assay (tq-PCR) in a single closed tube for differential detection and quantitative analysis of HAdV serotypes 2, 3 and 7. The sensitivity, specificity, reproducibility and clinical performance of tq-PCR were evaluated. The analytical sensitivity of the tq-PCR was 100 copies/reaction for each of HAdV serotypes 2, 3 and 7, and no cross-reaction with other common respiratory viruses or HAdV serotypes 1,4,5,6,31,55 and 57 was observed. The coefficients of variation (CV) of intra-assay and inter-assay were between 0.6% to 3.6%. Of 138 previously-defined HAdV-positive nasopharyngeal aspirates samples tested, the detection agreement between tq-PCR and nested PCR was 96.38% (133/138). The proposed tq-PCR assay is a sensitive, specific and reproducible method and has the potential for clinical use in the rapid and differential detection and quantitation of HAdV serotypes 2, 3 and 7.

  19. Methane, Ethane, and Propane Sensor for Real-time Leak Detection and Diagnostics

    Energy Technology Data Exchange (ETDEWEB)

    Roscioli, Joseph R. [Aerodyne Research, Inc., Billerica, MA (United States); Herndon, Scott [Aerodyne Research, Inc., Billerica, MA (United States); Nelson, David D. [Aerodyne Research, Inc., Billerica, MA (United States); Yacovitch, Tara [Aerodyne Research, Inc., Billerica, MA (United States)

    2017-03-24

    monitor was designed based upon the commercial compact mini-spectrometer capable of dual-laser operation. We intend to build and test this during phase II. Multiple opportunities for improvement were also identified. First, the reported ethane and propane concentrations are susceptible to external acceleration acting upon the instrument. During phase II we will address this “motion-sickness”. Second, significant software development will be needed operate the monitor at 1 second resolution in real time, and provide rapid, actionable data to a driver or passenger.

  20. Real time damage detection using recursive principal components and time varying auto-regressive modeling

    Science.gov (United States)

    Krishnan, M.; Bhowmik, B.; Hazra, B.; Pakrashi, V.

    2018-02-01

    experiments performed on a cantilever beam subjected to earthquake excitation; a two-storey benchscale model with a TMD and, data from recorded responses of UCLA factor building demonstrate the efficacy of the proposed methodology as an ideal candidate for real time, reference free structural health monitoring.

  1. Development of a surface plasmon resonance biosensor for real-time detection of osteogenic differentiation in live mesenchymal stem cells.

    Directory of Open Access Journals (Sweden)

    Yi-Chun Kuo

    Full Text Available Surface plasmon resonance (SPR biosensors have been recognized as a useful tool and widely used for real-time dynamic analysis of molecular binding affinity because of its high sensitivity to the change of the refractive index of tested objects. The conventional methods in molecular biology to evaluate cell differentiation require cell lysis or fixation, which make investigation in live cells difficult. In addition, a certain amount of cells are needed in order to obtain adequate protein or messenger ribonucleic acid for various assays. To overcome this limitation, we developed a unique SPR-based biosensing apparatus for real-time detection of cell differentiation in live cells according to the differences of optical properties of the cell surface caused by specific antigen-antibody binding. In this study, we reported the application of this SPR-based system to evaluate the osteogenic differentiation of mesenchymal stem cells (MSCs. OB-cadherin expression, which is up-regulated during osteogenic differentiation, was targeted under our SPR system by conjugating antibodies against OB-cadherin on the surface of the object. A linear relationship between the duration of osteogenic induction and the difference in refractive angle shift with very high correlation coefficient was observed. To sum up, the SPR system and the protocol reported in this study can rapidly and accurately define osteogenic maturation of MSCs in a live cell and label-free manner with no need of cell breakage. This SPR biosensor will facilitate future advances in a vast array of fields in biomedical research and medical diagnosis.

  2. Quantum dots-hyperbranched polyether hybrid nanospheres towards delivery and real-time detection of nitric oxide

    DEFF Research Database (Denmark)

    Liu, Shuiping; Gu, Tianxun; Fu, Jiajia

    2014-01-01

    In this work, novel hybrid nanosphere vehicles were synthesized for nitric oxide (NO) donating and real-time detection. The hybrid nanosphere vehicles consist of cadmium selenide quantum dots (CdSe QDs) as NO fluorescent probes, and the modified hyperbranched polyether (mHP)-based diazeniumdiolates...... as NO donors, respectively. The nanospheres have spherical outline with dimension of ~ 127 nm. The data of systematic characterization demonstrated that the mHP-based hybrid nanosphere vehicles (QDs-mHP-NO) can release and real-time detect NO with the low limit of 25 nM, based on fluorescence quenching...

  3. Real time detection of exhaled human breath using quantum cascade laser based sensor technology

    Science.gov (United States)

    Tittel, Frank K.; Lewicki, Rafal; Dong, Lei; Liu, Kun; Risby, Terence H.; Solga, Steven; Schwartz, Tim

    2012-02-01

    The development and performance of a cw, TE-cooled DFB quantum cascade laser based sensor for quantitative measurements of ammonia (NH3) and nitric oxide (NO) concentrations present in exhaled breath will be reported. Human breath contains ~ 500 different chemical species, usually at ultra low concentration levels, which can serve as biomarkers for the identification and monitoring of human diseases or wellness states. By monitoring NH3 concentration levels in exhaled breath a fast, non-invasive diagnostic method for treatment of patients with liver and kidney disorders, is feasible. The NH3 concentration measurements were performed with a 2f wavelength modulation quartz enhanced photoacoustic spectroscopy (QEPAS) technique, which is suitable for real time breath measurements, due to the fast gas exchange inside a compact QEPAS gas cell. A Hamamatsu air-cooled high heat load (HHL) packaged CW DFB-QCL is operated at 17.5°C, targeting the optimum interference free NH3 absorption line at 967.35 cm-1 (λ~10.34 μm), with ~ 20 mW of optical power. The sensor architecture includes a reference cell, filled with a 2000 ppmv NH3 :N2 mixture at 130 Torr, which is used for absorption line-locking. A minimum detection limit (1σ) for the line locked NH3 sensor is ~ 6 ppbv (with a 1σ 1 sec time resolution of the control electronics). This NH3 sensor was installed in late 2010 and is being clinically tested at St. Luke's Hospital in Bethlehem, PA.

  4. Socio-economic Impact Analysis for Near Real-Time Flood Detection in the Lower Mekong River Basin

    Science.gov (United States)

    Oddo, P.; Ahamed, A.; Bolten, J. D.

    2017-12-01

    Flood events pose a severe threat to communities in the Lower Mekong River Basin. The combination of population growth, urbanization, and economic development exacerbate the impacts of these flood events. Flood damage assessments are frequently used to quantify the economic losses in the wake of storms. These assessments are critical for understanding the effects of flooding on the local population, and for informing decision-makers about future risks. Remote sensing systems provide a valuable tool for monitoring flood conditions and assessing their severity more rapidly than traditional post-event evaluations. The frequency and severity of extreme flood events are projected to increase, further illustrating the need for improved flood monitoring and impact analysis. In this study we implement a socio-economic damage model into a decision support tool with near real-time flood detection capabilities (NASA's Project Mekong). Surface water extent for current and historical floods is found using multispectral Moderate-resolution Imaging Spectroradiometer (MODIS) 250-meter imagery and the spectral Normalized Difference Vegetation Index (NDVI) signatures of permanent water bodies (MOD44W). Direct and indirect damages to populations, infrastructure, and agriculture are assessed using the 2011 Southeast Asian flood as a case study. Improved land cover and flood depth assessments result in a more refined understanding of losses throughout the Mekong River Basin. Results suggest that rapid initial estimates of flood impacts can provide valuable information to governments, international agencies, and disaster responders in the wake of extreme flood events.

  5. Real-time, high frequency QRS electrocardiograph with reduced amplitude zone detection

    Science.gov (United States)

    Schlegel, Todd T. (Inventor); DePalma, Jude L. (Inventor); Moradi, Saeed (Inventor)

    2009-01-01

    Real time cardiac electrical data are received from a patient, manipulated to determine various useful aspects of the ECG signal, and displayed in real time in a useful form on a computer screen or monitor. The monitor displays the high frequency data from the QRS complex in units of microvolts, juxtaposed with a display of conventional ECG data in units of millivolts or microvolts. The high frequency data are analyzed for their root mean square (RMS) voltage values and the discrete RMS values and related parameters are displayed in real time. The high frequency data from the QRS complex are analyzed with imbedded algorithms to determine the presence or absence of reduced amplitude zones, referred to herein as ''RAZs''. RAZs are displayed as ''go, no-go'' signals on the computer monitor. The RMS and related values of the high frequency components are displayed as time varying signals, and the presence or absence of RAZs may be similarly displayed over time.

  6. Interlaboratory validation data on real-time polymerase chain reaction detection for unauthorized genetically modified papaya line PRSV-YK

    Directory of Open Access Journals (Sweden)

    Kosuke Nakamura

    2016-06-01

    Real-time polymerase chain reaction (PCR detection method for unauthorized genetically modified (GM papaya (Carica papaya L. line PRSV-YK (PRSV-YK detection method was developed using whole genome sequence data (DDBJ Sequenced Read Archive under accession No. PRJDB3976. Interlaboratory validation datasets for PRSV-YK detection method were provided. Data indicating homogeneity of samples prepared for interlaboratory validation were included. Specificity and sensitivity test data for PRSV-YK detection method were also provided.

  7. Real time 1.55 μm VCSEL-based coherent detection link

    DEFF Research Database (Denmark)

    Rodes Lopez, Roberto; Parekh, D.; Jensen, Jesper Bevensee

    2012-01-01

    This paper presents an experimental demonstration of VCSEL-based PON with simplified real-time coherent receiver at 2.5 Gbps. Receiver sensitivity of −37 dBm is achieved proving splitting ratio up to 2048 after 17 km fiber transmission.......This paper presents an experimental demonstration of VCSEL-based PON with simplified real-time coherent receiver at 2.5 Gbps. Receiver sensitivity of −37 dBm is achieved proving splitting ratio up to 2048 after 17 km fiber transmission....

  8. Comparison of DNA extraction kits for detection of Burkholderia pseudomallei in spiked human whole blood using real-time PCR.

    Directory of Open Access Journals (Sweden)

    Nicole L Podnecky

    Full Text Available Burkholderia pseudomallei, the etiologic agent of melioidosis, is endemic in northern Australia and Southeast Asia and can cause severe septicemia that may lead to death in 20% to 50% of cases. Rapid detection of B. pseudomallei infection is crucial for timely treatment of septic patients. This study evaluated seven commercially available DNA extraction kits to determine the relative recovery of B. pseudomallei DNA from spiked EDTA-containing human whole blood. The evaluation included three manual kits: the QIAamp DNA Mini kit, the QIAamp DNA Blood Mini kit, and the High Pure PCR Template Preparation kit; and four automated systems: the MagNAPure LC using the DNA Isolation Kit I, the MagNAPure Compact using the Nucleic Acid Isolation Kit I, and the QIAcube using the QIAamp DNA Mini kit and the QIAamp DNA Blood Mini kit. Detection of B. pseudomallei DNA extracted by each kit was performed using the B. pseudomallei specific type III secretion real-time PCR (TTS1 assay. Crossing threshold (C T values were used to compare the limit of detection and reproducibility of each kit. This study also compared the DNA concentrations and DNA purity yielded for each kit. The following kits consistently yielded DNA that produced a detectable signal from blood spiked with 5.5×10(4 colony forming units per mL: the High Pure PCR Template Preparation, QIAamp DNA Mini, MagNA Pure Compact, and the QIAcube running the QIAamp DNA Mini and QIAamp DNA Blood Mini kits. The High Pure PCR Template Preparation kit yielded the lowest limit of detection with spiked blood, but when this kit was used with blood from patients with confirmed cases of melioidosis, the bacteria was not reliably detected indicating blood may not be an optimal specimen.

  9. Rapid process development of chromatographic process using direct analysis in real time mass spectrometry as a process analytical technology tool.

    Science.gov (United States)

    Yan, Binjun; Chen, Teng; Xu, Zhilin; Qu, Haibin

    2014-06-01

    The concept of quality by design (QbD) is widely applied in the process development of pharmaceuticals. However, the additional cost and time have caused some resistance about QbD implementation. To show a possible solution, this work proposed a rapid process development method, which used direct analysis in real time mass spectrometry (DART-MS) as a process analytical technology (PAT) tool for studying the chromatographic process of Ginkgo biloba L., as an example. The breakthrough curves were fast determined by DART-MS at-line. A high correlation coefficient of 0.9520 was found between the concentrations of ginkgolide A determined by DART-MS and HPLC. Based on the PAT tool, the impacts of process parameters on the adsorption capacity were discovered rapidly, which showed a decreased adsorption capacity with the increase of the flow rate. This work has shown the feasibility and advantages of integrating PAT into QbD implementation for rapid process development. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Real-time PCR for the early detection and quantification of Coxiella burnetii as an alternative to the murine bioassay.

    Science.gov (United States)

    Howe, Gerald B; Loveless, Bonnie M; Norwood, David; Craw, Philip; Waag, David; England, Marilyn; Lowe, John R; Courtney, Bernard C; Pitt, M Louise; Kulesh, David A

    2009-01-01

    Real-time PCR was used to analyze archived blood from non-human primates (NHP) and fluid samples originating from a well-controlled Q fever vaccine efficacy trial. The PCR targets were the IS1111 element and the com1 gene of Coxiella burnetii. Data from that previous study were used to evaluate real-time PCR as an alternative to the use of sero-conversion by mouse bioassay for both quantification and early detection of C. burnetii bacteria. Real-time PCR and the mouse bioassay exhibited no statistical difference in quantifying the number of microorganisms delivered in the aerosol challenge dose. The presence of C. burnetii in peripheral blood of non-human primates was detected by real-time PCR as early after exposure as the mouse bioassay with results available within hours instead of weeks. This study demonstrates that real-time PCR has the ability to replace the mouse bioassay to measure dosage and monitor infection of C. burnetii in a non-human primate model.

  11. European validation of a real-time PCR-based method for detection of Listeria monocytogenes in soft cheese.

    Science.gov (United States)

    Gianfranceschi, Monica Virginia; Rodriguez-Lazaro, David; Hernandez, Marta; González-García, Patricia; Comin, Damiano; Gattuso, Antonietta; Delibato, Elisabetta; Sonnessa, Michele; Pasquali, Frederique; Prencipe, Vincenza; Sreter-Lancz, Zuzsanna; Saiz-Abajo, María-José; Pérez-De-Juan, Javier; Butrón, Javier; Kozačinski, Lidija; Tomic, Danijela Horvatek; Zdolec, Nevijo; Johannessen, Gro S; Jakočiūnė, Džiuginta; Olsen, John Elmerdahl; De Santis, Paola; Lovari, Sarah; Bertasi, Barbara; Pavoni, Enrico; Paiusco, Antonella; De Cesare, Alessandra; Manfreda, Gerardo; De Medici, Dario

    2014-08-01

    The classical microbiological method for detection of Listeria monocytogenes requires around 7 days for final confirmation, and due to perishable nature of RTE food products, there is a clear need for an alternative methodology for detection of this pathogen. This study presents an international (at European level) ISO 16140-based validation trial of a non-proprietary real-time PCR-based methodology that can generate final results in the following day of the analysis. This methodology is based on an ISO compatible enrichment coupled to a bacterial DNA extraction and a consolidated real-time PCR assay. Twelve laboratories from six European countries participated in this trial, and soft cheese was selected as food model since it can represent a difficult matrix for the bacterial DNA extraction and real-time PCR amplification. The limit of detection observed was down to 10 CFU per 25 of sample, showing excellent concordance and accordance values between samples and laboratories (>75%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (82.75%, 96.70% and 97.62%, respectively) when the results obtained for the real-time PCR-based methods were compared to those of the ISO 11290-1 standard method. An interesting observation was that the L. monocytogenes detection by the real-time PCR method was less affected in the presence of Listeria innocua in the contaminated samples, proving therefore to be more reliable than the reference method. The results of this international trial demonstrate that the evaluated real-time PCR-based method represents an excellent alterative to the ISO standard since it shows a higher performance as well as reduce the extent of the analytical process, and can be easily implemented routinely by the competent authorities and food industry laboratories. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Multiplex real-time PCR for detection of Staphylococcus aureus, mecA and Panton-Valentine Leukocidin (PVL genes from selective enrichments from animals and retail meat.

    Directory of Open Access Journals (Sweden)

    Valeria Velasco

    Full Text Available The aim of this study was to compare a real-time PCR assay, with a conventional culture/PCR method, to detect S. aureus, mecA and Panton-Valentine Leukocidin (PVL genes in animals and retail meat, using a two-step selective enrichment protocol. A total of 234 samples were examined (77 animal nasal swabs, 112 retail raw meat, and 45 deli meat. The multiplex real-time PCR targeted the genes: nuc (identification of S. aureus, mecA (associated with methicillin resistance and PVL (virulence factor, and the primary and secondary enrichment samples were assessed. The conventional culture/PCR method included the two-step selective enrichment, selective plating, biochemical testing, and multiplex PCR for confirmation. The conventional culture/PCR method recovered 95/234 positive S. aureus samples. Application of real-time PCR on samples following primary and secondary enrichment detected S. aureus in 111/234 and 120/234 samples respectively. For detection of S. aureus, the kappa statistic was 0.68-0.88 (from substantial to almost perfect agreement and 0.29-0.77 (from fair to substantial agreement for primary and secondary enrichments, using real-time PCR. For detection of mecA gene, the kappa statistic was 0-0.49 (from no agreement beyond that expected by chance to moderate agreement for primary and secondary enrichment samples. Two pork samples were mecA gene positive by all methods. The real-time PCR assay detected the mecA gene in samples that were negative for S. aureus, but positive for Staphylococcus spp. The PVL gene was not detected in any sample by the conventional culture/PCR method or the real-time PCR assay. Among S. aureus isolated by conventional culture/PCR method, the sequence type ST398, and multi-drug resistant strains were found in animals and raw meat samples. The real-time PCR assay may be recommended as a rapid method for detection of S. aureus and the mecA gene, with further confirmation of methicillin-resistant S. aureus (MRSA

  13. Multiplex Real-Time PCR for Detection of Staphylococcus aureus, mecA and Panton-Valentine Leukocidin (PVL) Genes from Selective Enrichments from Animals and Retail Meat

    Science.gov (United States)

    Velasco, Valeria; Sherwood, Julie S.; Rojas-García, Pedro P.; Logue, Catherine M.

    2014-01-01

    The aim of this study was to compare a real-time PCR assay, with a conventional culture/PCR method, to detect S. aureus, mecA and Panton-Valentine Leukocidin (PVL) genes in animals and retail meat, using a two-step selective enrichment protocol. A total of 234 samples were examined (77 animal nasal swabs, 112 retail raw meat, and 45 deli meat). The multiplex real-time PCR targeted the genes: nuc (identification of S. aureus), mecA (associated with methicillin resistance) and PVL (virulence factor), and the primary and secondary enrichment samples were assessed. The conventional culture/PCR method included the two-step selective enrichment, selective plating, biochemical testing, and multiplex PCR for confirmation. The conventional culture/PCR method recovered 95/234 positive S. aureus samples. Application of real-time PCR on samples following primary and secondary enrichment detected S. aureus in 111/234 and 120/234 samples respectively. For detection of S. aureus, the kappa statistic was 0.68–0.88 (from substantial to almost perfect agreement) and 0.29–0.77 (from fair to substantial agreement) for primary and secondary enrichments, using real-time PCR. For detection of mecA gene, the kappa statistic was 0–0.49 (from no agreement beyond that expected by chance to moderate agreement) for primary and secondary enrichment samples. Two pork samples were mecA gene positive by all methods. The real-time PCR assay detected the mecA gene in samples that were negative for S. aureus, but positive for Staphylococcus spp. The PVL gene was not detected in any sample by the conventional culture/PCR method or the real-time PCR assay. Among S. aureus isolated by conventional culture/PCR method, the sequence type ST398, and multi-drug resistant strains were found in animals and raw meat samples. The real-time PCR assay may be recommended as a rapid method for detection of S. aureus and the mecA gene, with further confirmation of methicillin-resistant S. aureus (MRSA) using

  14. Label-free detection of gliadin food allergen mediated by real-time apta-PCR.

    Science.gov (United States)

    Pinto, Alessandro; Polo, Pedro Nadal; Henry, Olivier; Redondo, M Carmen Bermudo; Svobodova, Marketa; O'Sullivan, Ciara K

    2014-01-01

    Celiac disease is an immune-mediated enteropathy triggered by the ingestion of gluten. The only effective treatment consists in a lifelong gluten-free diet, requiring the food industry to tightly control the gluten contents of their products. To date, several gluten quantification approaches using antibodies are available and recommended by the legal authorities, such as Codex Alimentarius. However, whilst these antibody-based tests exhibit high sensitivity and specificity, the production of antibodies inherently requires the killing of host animals and is time-consuming and relatively expensive. Aptamers are structured single-stranded nucleic acid ligands that bind with high affinity and specificity to their cognate target, and aiming for a cost-effective viable alternative to the use of antibodies. Herein, we report the systematic evolution of ligands by exponential enrichment (SELEX)-based selection of a DNA aptamer against gliadin from a combinatorial DNA library and its application in a novel detection assay. Taking into account the hydrophobic nature of the gliadin target, a microtitre plate format was exploited for SELEX, where the target was immobilised via hydrophobic interactions, thus exposing aptatopes accessible for interaction with the DNA library. Evolution was followed using surface plasmon resonance, and following eight rounds of SELEX, the enriched DNA pool was cloned, sequenced and a clear consensus motif was identified. An apta-PCR assay was developed where competition for the aptamer takes place between the surface-immobilised gliadin and gliadin in the target sample, akin to an ELISA competitive format where the more target present in the sample, the less aptamer will bind to the immobilised gliadin. Following competition, any aptamer bound to the immobilised gliadin was heat-eluted and quantitatively amplified using real-time PCR, achieving a detection limit of approx. 2 nM (100 ng mL(-1)). The specificity of the selected aptamer was

  15. Two quantitative real-time PCR assays for the detection of penaeid shrimp and blue crab, crustacean shellfish allergens.

    Science.gov (United States)

    Eischeid, Anne C; Kim, Bang-hyun; Kasko, Sasha M

    2013-06-19

    Food allergen detection methods must be able to specifically detect minute quantities of an allergenic food in a complex food matrix. One technique that can be used is real-time PCR. For the work described here, real-time PCR assays were developed to detect penaeid shrimp and blue crab, crustacean shellfish allergens. The method was tested using shrimp meat and crab meat spiked into several types of foods, including canned soups, deli foods, meat, seafood, and prepared seafood products. Foods were spiked with either shrimp or crab at levels ranging from 0.1 to 10⁶ parts per million (ppm) and analyzed either raw or cooked by a variety of methods. Real-time PCR data were used to generate linear standard curves, and assays were evaluated with respect to linear range and reaction efficiency. Results indicate that both assays performed well in a variety of food types. High reaction efficiencies were achieved across a linear range of 6-8 orders of magnitude. Limits of detection were generally between 0.1 and 1 ppm. Cooking methods used to simulate thermal processing of foods had little effect on assay performance. This work demonstrates that real-time PCR can be a valuable tool in the detection of crustacean shellfish.

  16. A timed-automata approach for critical path detection in a soft real-time application

    NARCIS (Netherlands)

    Yildiz, Bugra Mehmet; Bockisch, Christoph; Rensink, Arend; Aksit, Mehmet

    In this paper, we report preliminary ideas from our project called “Time Performance Improvement With Parallel Processing Systems‿ (TIPS). In the TIPS project, we plan to take advantage of multi-core platforms for performance improvement by parallelizing a complex soft real-time application. In

  17. Detection of Fusobacterium necrophorum subsp funduliforme in tonsillitis in young adults by real-time PCR

    DEFF Research Database (Denmark)

    Jensen, Anders; Hagelskjær Kristensen, Lena; Prag, Jørgen

    2007-01-01

    Throat swabs from 61 patients, aged 18-32 years, with non-streptococcal tonsillitis (NST) and 92 healthy controls were examined for the presence of Fusobacterium necrophorum DNA using a novel TaqMan-based real-time quantitative PCR assay for F. necrophorum subspecies. The assay was based on the g...

  18. Real-time MEG neurofeedback training of posterior alpha activity modulates subsequent visual detection performance

    NARCIS (Netherlands)

    Okazaki, Y.O.; Horschig, J.; Luther, L.M.; Oostenveld, R.; Murakami, I.; Jensen, O.

    2015-01-01

    It has been demonstrated that alpha activity is lateralized when attention is directed to the left or right visual hemifield. We investigated whether real-time neurofeedback training of the alpha lateralization enhances participants' ability to modulate posterior alpha lateralization and causes

  19. Real time quantitative amplification detection on a microarray: towards high multiplex quantitative PCR.

    NARCIS (Netherlands)

    Pierik, A.; Moamfa, M; van Zelst, M.; Clout, D.; Stapert, H.; Dijksman, Johan Frederik; Broer, D.; Wimberger-Friedl, R.

    2012-01-01

    Quantitative real-time polymerase chain reaction (qrtPCR) is widely used as a research and diagnostic tool. Notwithstanding its many powerful features, the method is limited in the degree of multiplexing to about 6 due to spectral overlap of the available fluorophores. A new method is presented that

  20. Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR

    NARCIS (Netherlands)

    Pierik, Anke; Boamfa, M.; Zelst, van M.; Clout, D.; Stapert, H.R.; Dijksman, J.F.; Broer, D.J.; Wimberger-Friedl, R.

    2012-01-01

    Quantitative real-time polymerase chain reaction (qrtPCR) is widely used as a research and diagnostic tool. Notwithstanding its many powerful features, the method is limited in the degree of multiplexing to about 6 due to spectral overlap of the available fluorophores. A new method is presented that

  1. Detection of Legionella pneumophila by real-time PCR for the mip gene.

    Science.gov (United States)

    Wilson, Deborah A; Yen-Lieberman, Belinda; Reischl, Udo; Gordon, Steve M; Procop, Gary W

    2003-07-01

    A real-time PCR assay for the mip gene of Legionella pneumophila was tested with 27 isolates of L. pneumophila, 20 isolates of 14 other Legionella species, and 103 non-Legionella bacteria. Eight culture-positive and 40 culture-negative clinical specimens were tested. This assay was 100% sensitive and 100% specific for L. pneumophila.

  2. Real-time PCR assays for detection of Brucella spp. and the identification of genotype ST27 in bottlenose dolphins (Tursiops truncatus).

    Science.gov (United States)

    Wu, Qingzhong; McFee, Wayne E; Goldstein, Tracey; Tiller, Rebekah V; Schwacke, Lori

    2014-05-01

    Rapid detection of Brucella spp. in marine mammals is challenging. Microbiologic culture is used for definitive diagnosis of brucellosis, but is time consuming, has low sensitivity and can be hazardous to laboratory personnel. Serological methods can aid in diagnosis, but may not differentiate prior exposure versus current active infection and may cross-react with unrelated Gram-negative bacteria. This study reports a real-time PCR assay for the detection of Brucella spp. and application to screen clinical samples from bottlenose dolphins stranded along the coast of South Carolina, USA. The assay was found to be 100% sensitive for the Brucella strains tested, and the limit of detection was 0.27fg of genomic DNA from Brucella ceti B1/94 per PCR volume. No amplification was detected for the non-Brucella pathogens tested. Brucella DNA was detected in 31% (55/178) of clinical samples tested. These studies indicate that the real-time PCR assay is highly sensitive and specific for the detection of Brucella spp. in bottlenose dolphins. We also developed a second real-time PCR assay for rapid identification of Brucella ST27, a genotype that is associated with human zoonotic infection. Positive results were obtained for Brucella strains which had been identified as ST27 by multilocus sequence typing. No amplification was found for other Brucella strains included in this study. ST27 was identified in 33% (18/54) of Brucella spp. DNA-positive clinical samples. To our knowledge, this is the first report on the use of a real-time PCR assay for identification of Brucella genotype ST27 in marine mammals. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Flood Foresight: A near-real time flood monitoring and forecasting tool for rapid and predictive flood impact assessment

    Science.gov (United States)

    Revilla-Romero, Beatriz; Shelton, Kay; Wood, Elizabeth; Berry, Robert; Bevington, John; Hankin, Barry; Lewis, Gavin; Gubbin, Andrew; Griffiths, Samuel; Barnard, Paul; Pinnell, Marc; Huyck, Charles

    2017-04-01

    The hours and days immediately after a major flood event are often chaotic and confusing, with first responders rushing to mobilise emergency responders, provide alleviation assistance and assess loss to assets of interest (e.g., population, buildings or utilities). Preparations in advance of a forthcoming event are becoming increasingly important; early warning systems have been demonstrated to be useful tools for decision markers. The extent of damage, human casualties and economic loss estimates can vary greatly during an event, and the timely availability of an accurate flood extent allows emergency response and resources to be optimised, reduces impacts, and helps prioritise recovery. In the insurance sector, for example, insurers are under pressure to respond in a proactive manner to claims rather than waiting for policyholders to report losses. Even though there is a great demand for flood inundation extents and severity information in different sectors, generating flood footprints for large areas from hydraulic models in real time remains a challenge. While such footprints can be produced in real time using remote sensing, weather conditions and sensor availability limit their ability to capture every single flood event across the globe. In this session, we will present Flood Foresight (www.floodforesight.com), an operational tool developed to meet the universal requirement for rapid geographic information, before, during and after major riverine flood events. The tool provides spatial data with which users can measure their current or predicted impact from an event - at building, basin, national or continental scales. Within Flood Foresight, the Screening component uses global rainfall predictions to provide a regional- to continental-scale view of heavy rainfall events up to a week in advance, alerting the user to potentially hazardous situations relevant to them. The Forecasting component enhances the predictive suite of tools by providing a local

  4. CNR considerations for rapid real-time MRI tumor tracking in radiotherapy hybrid devices: Effects of B0 field strength

    International Nuclear Information System (INIS)

    Wachowicz, K.; De Zanche, N.; Yip, E.; Volotovskyy, V.; Fallone, B. G.

    2016-01-01

    Purpose: This work examines the subject of contrast-to-noise ratio (CNR), specifically between tumor and tissue background, and its dependence on the MRI field strength, B 0 . This examination is motivated by the recent interest and developments in MRI/radiotherapy hybrids where real-time imaging can be used to guide treatment beams. The ability to distinguish a tumor from background tissue is of primary importance in this field, and this work seeks to elucidate the complex relationship between the CNR and B 0 that is too often assumed to be purely linear. Methods: Experimentally based models of B 0 -dependant relaxation for various tumor and normal tissues from the literature were used in conjunction with signal equations for MR sequences suitable for rapid real-time imaging to develop field-dependent predictions for CNR. These CNR models were developed for liver, lung, breast, glioma, and kidney tumors for spoiled gradient-echo, balanced steady-state free precession (bSSFP), and single-shot half-Fourier fast spin echo sequences. Results: Due to the pattern in which the relaxation properties of tissues are found to vary over B 0 field (specifically the T 1 time), there was always an improved CNR at lower fields compared to linear dependency. Further, in some tumor sites, the CNR at lower fields was found to be comparable to, or sometimes higher than those at higher fields (i.e., bSSFP CNR for glioma, kidney, and liver tumors). Conclusions: In terms of CNR, lower B 0 fields have been shown to perform as well or better than higher fields for some tumor sites due to superior T 1 contrast. In other sites this effect was less pronounced, reversing the CNR advantage. This complex relationship between CNR and B 0 reveals both low and high magnetic fields as viable options for tumor tracking in MRI/radiotherapy hybrids.

  5. Multiplex real-time PCR (TaqMan) assay for the simultaneous detection and discrimination of potato powdery and common scab diseases and pathogens.

    Science.gov (United States)

    Qu, X S; Wanner, L A; Christ, B J

    2011-03-01

    To develop a multiplex real-time PCR assay using TaqMan probes for the simultaneous detection and discrimination of potato powdery scab and common scab, two potato tuber diseases with similar symptoms, and the causal pathogens Spongospora subterranea and plant pathogenic Streptomyces spp. Real-time PCR primers and a probe for S. subterranea were designed based on the DNA sequence of the ribosomal RNA ITS2 region. Primers and a probe for pathogenic Streptomyces were designed based on the DNA sequence of the txtAB genes. The two sets of primer pairs and probes were used in a single real-time PCR assay. The multiplex real-time PCR assay was confirmed to be specific for S. subterranea and pathogenic Streptomyces. The assay detected DNA quantities of 100 fg for each of the two pathogens and linear responses and high correlation coefficients between the amount of DNA and C(t) values for each pathogen were achieved. The presence of two sets of primer pairs and probes and of plant extracts did not alter the sensitivity and efficiency of multiplex PCR amplification. Using the PCR assay, we could discriminate between powdery scab and common scab tubers with similar symptoms. Common scab and powdery scab were detected in some tubers with no visible symptoms. Mixed infections of common scab and powdery scab on single tubers were also revealed. This multiplex real-time PCR assay is a rapid, cost efficient, specific and sensitive tool for the simultaneous detection and discrimination of the two pathogens on infected potato tubers when visual symptoms are inconclusive or not present. Accurate and quick identification and discrimination of the cause of scab diseases on potatoes will provide critical information to potato growers and researchers for disease management. This is important because management strategies for common and powdery scab diseases are very different. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  6. Increased detection of mastitis pathogens by real-time PCR compared to bacterial culture.

    Science.gov (United States)

    Keane, O M; Budd, K E; Flynn, J; McCoy, F

    2013-09-21

    Rapid and accurate identification of mastitis pathogens is important for disease control. Bacterial culture and isolate identification is considered the gold standard in mastitis diagnosis but is time consuming and results in many culture-negative samples. Identification of mastitis pathogens by PCR has been proposed as a fast and sensitive alternative to bacterial culture. The results of bacterial culture and PCR for the identification of the aetiological agent of clinical mastitis were compared. The pathogen identified by traditional culture methods was also detected by PCR in 98 per cent of cases indicating good agreement between the positive results of bacterial culture and PCR. A mastitis pathogen could not be recovered from approximately 30 per cent of samples by bacterial culture, however, an aetiological agent was identified by PCR in 79 per cent of these samples. Therefore, a mastitis pathogen was detected in significantly more milk samples by PCR than by bacterial culture (92 per cent and 70 per cent, respectively) although the clinical relevance of PCR-positive culture-negative results remains controversial. A mixed infection of two or more mastitis pathogens was also detected more commonly by PCR. Culture-negative samples due to undetected Staphylococcus aureus infections were rare. The use of PCR technology may assist in rapid mastitis diagnosis, however, accurate interpretation of PCR results in the absence of bacterial culture remains problematic.

  7. Mycoplasma detection by triplex real-time PCR in bronchoalveolar lavage fluid from bovine respiratory disease complex cases

    NARCIS (Netherlands)

    Cornelissen, Jan B.W.J.; Bree, de Freddy M.; Wal, van der Fimme J.; Kooij, Engbert A.; Koene, Miriam G.J.; Bossers, Alex; Smid, Bregtje; Antonis, Adriaan F.; Wisselink, Henk J.

    2017-01-01

    Background: In this study we evaluated the RespoCheck Mycoplasma triplex real-time PCR for the detection in bronchoalveolar lavage fluid (BALF) of Mycoplasma (M.) dispar, M. bovis and M. bovirhinis, all three associated with bovine respiratory disease (BRD). Primers and probes of the RespoCheck

  8. Combination of real-time PCR and sequencing to detect multiple clinically relevant genetic variations in the lactase gene

    DEFF Research Database (Denmark)

    Brasen, Claus Lohman; Frischknecht, Lone; Ørnskov, Dorthe

    2017-01-01

    in the probe-binding site may cause errors in analysis. The aim of this study was to determine the prevalence of the variants in a Danish cohort examined for lactose intolerance as well as to improve the real-time PCR analysis for detection of the different variants. METHODS: We genotyped 3395 routine samples...

  9. Partial validation of a TaqMan real-time quantitative PCR for the detection of ranaviruses

    DEFF Research Database (Denmark)

    Stilwell, Natalie K.; Whittington, Richard J.; Hick, Paul M.

    2018-01-01

    Ranaviruses are globally emerging pathogens negatively impacting wild and cultured fish, amphibians, and reptiles. Although conventional and diagnostic real-time PCR (qPCR) assays have been developed to detect ranaviruses, these assays often have not been tested against the known diversity of ran...

  10. Development of a real-time absorption method for detecting the mercaptan odorizing mixture of natural gas

    NARCIS (Netherlands)

    Kireev, SV; Petrov, NG; Podolyako, EM; Shnyrev, SL

    The absorption of mercaptan mixtures used for odorizing natural gas and mixtures of natural gas is experimentally studied in the spectral range 2.5-20 mu m. An absorption method for the real-time detection of the odorant concentration is proposed. The method is based on intensity measurements of the

  11. Comparison of different real-time PCR chemistries and their suitability for detection and quantification of genetically modified organisms

    NARCIS (Netherlands)

    Gasparic, M.B.; Cankar, K.; Zel, J.; Gruden, K.

    2008-01-01

    Background: The real-time polymerase chain reaction is currently the method of choice for quantifying nucleic acids in different DNA based quantification applications. It is widely used also for detecting and quantifying genetically modified components in food and feed, predominantly employing

  12. Development of Candida-Specific Real-Time PCR Assays for the Detection and Identification of Eight Medically Important Candida Species.

    Science.gov (United States)

    Zhang, Jing; Hung, Guo-Chiuan; Nagamine, Kenjiro; Li, Bingjie; Tsai, Shien; Lo, Shyh-Ching

    2016-01-01

    Culture-based identification methods have been the gold standard for the diagnosis of fungal infection. Currently, molecular technologies such as real-time PCR assays with short turnaround time can provide desirable alternatives for the rapid detection of Candida microbes. However, most of the published PCR primer sets are not Candida specific and likely to amplify DNA from common environmental contaminants, such as Aspergillus microbes. In this study, we designed pan-Candida primer sets based on the ribosomal DNA-coding regions conserved within Candida but distinct from those of Aspergillus and Penicillium. We demonstrate that the final two selected pan-Candida primer sets would not amplify Aspergillus DNA and could be used to differentiate eight medically important Candida pathogens in real-time PCR assays based on their melting profiles, with a sensitivity of detection as low as 10 fg of Candida genomic DNA. Moreover, we further evaluated and selected species-specific primer sets covering Candida albicans, Candida glabrata, Candida tropicalis, and Candida dubliniensis and show that they had high sensitivity and specificity. These real-time PCR primer sets could potentially be assembled into a single PCR array for the rapid detection of Candida species in various clinical settings, such as corneal transplantation.

  13. Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR.

    Science.gov (United States)

    Mwaengo, D; Lorenzo, G; Iglesias, J; Warigia, M; Sang, R; Bishop, R P; Brun, A

    2012-10-01

    Diagnostic methods allowing for rapid identification of pathogens are crucial for controlling and preventing dissemination after disease outbreaks as well as for use in surveillance programs. For arboviruses, detection of the presence of virus in their arthropod hosts is important for monitoring of viral activity and quantitative information is useful for modeling of transmission dynamics. In this study, molecular detection of Rift Valley fever virus (RVFV) in mosquito samples from the 2006 to 2007 East African outbreaks was performed using quantitative real-time PCR assay (qRT-PCR). Specific RVFV sequence-based primer/fluorogenic (TaqMan) probe sets were derived from the L and S RNA segments of the virus. Both primer-probe L and S segment-based combinations detected genomic RVFV sequences, with generally comparable levels of sensitivity. Viral loads from three mosquito species, Aedes mcintoshi, Aedes ochraceus and Mansonia uniformis were estimated and significant differences of between 5- and 1000-fold were detected between Ae. mcintoshi and M. uniformis using both the L and S primer-probe-based assays. The genetic relationships of the viral sequences in mosquito samples were established by partial M segment sequencing and assigned to the two previously described viral lineages defined by analysis of livestock isolates obtained during the 2006-2007 outbreak, confirming that similar viruses were present in both the vector and mammalian host. The data confirms the utility of qRT-PCR for identification and initial quantification of virus in mosquito samples during RVFV outbreaks. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Direct Detection and Differentiation of Pathogenic Leptospira Species Using a Multi-Gene Targeted Real Time PCR Approach

    Science.gov (United States)

    Ferreira, Ana Sofia; Costa, Pedro; Rocha, Teresa; Amaro, Ana; Vieira, Maria Luísa; Ahmed, Ahmed; Thompson, Gertrude; Hartskeerl, Rudy A.; Inácio, João

    2014-01-01

    Leptospirosis is a growing public and veterinary health concern caused by pathogenic species of Leptospira. Rapid and reliable laboratory tests for the direct detection of leptospiral infections in animals are in high demand not only to improve diagnosis but also for understanding the epidemiology of the disease. In this work we describe a novel and simple TaqMan-based multi-gene targeted real-time PCR approach able to detect and differentiate Leptospira interrogans, L. kirschneri, L. borgpeteresenii and L. noguchii, which constitute the veterinary most relevant pathogenic species of Leptospira. The method uses sets of species-specific probes, and respective flanking primers, designed from ompL1 and secY gene sequences. To monitor the presence of inhibitors, a duplex amplification assay targeting both the mammal β-actin and the leptospiral lipL32 genes was implemented. The analytical sensitivity of all primer and probe sets was estimated to be <10 genome equivalents (GE) in the reaction mixture. Application of the amplification reactions on genomic DNA from a variety of pathogenic and non-pathogenic Leptospira strains and other non-related bacteria revealed a 100% analytical specificity. Additionally, pathogenic leptospires were successfully detected in five out of 29 tissue samples from animals (Mus spp., Rattus spp., Dolichotis patagonum and Sus domesticus). Two samples were infected with L. borgpetersenii, two with L. interrogans and one with L. kirschneri. The possibility to detect and identify these pathogenic agents to the species level in domestic and wildlife animals reinforces the diagnostic information and will enhance our understanding of the epidemiology of leptopirosis. PMID:25398140

  15. Detecting spatial patterns of rivermouth processes using a geostatistical framework for near-real-time analysis

    Science.gov (United States)

    Xu, Wenzhao; Collingsworth, Paris D.; Bailey, Barbara; Carlson Mazur, Martha L.; Schaeffer, Jeff; Minsker, Barbara

    2017-01-01

    This paper proposes a geospatial analysis framework and software to interpret water-quality sampling data from towed undulating vehicles in near-real time. The framework includes data quality assurance and quality control processes, automated kriging interpolation along undulating paths, and local hotspot and cluster analyses. These methods are implemented in an interactive Web application developed using the Shiny package in the R programming environment to support near-real time analysis along with 2- and 3-D visualizations. The approach is demonstrated using historical sampling data from an undulating vehicle deployed at three rivermouth sites in Lake Michigan during 2011. The normalized root-mean-square error (NRMSE) of the interpolation averages approximately 10% in 3-fold cross validation. The results show that the framework can be used to track river plume dynamics and provide insights on mixing, which could be related to wind and seiche events.

  16. Real-Time Polymerase Chain Reaction Detection of Angiostrongylus cantonensis DNA in Cerebrospinal Fluid from Patients with Eosinophilic Meningitis.

    Science.gov (United States)

    Qvarnstrom, Yvonne; Xayavong, Maniphet; da Silva, Ana Cristina Aramburu; Park, Sarah Y; Whelen, A Christian; Calimlim, Precilia S; Sciulli, Rebecca H; Honda, Stacey A A; Higa, Karen; Kitsutani, Paul; Chea, Nora; Heng, Seng; Johnson, Stuart; Graeff-Teixeira, Carlos; Fox, LeAnne M; da Silva, Alexandre J

    2016-01-01

    Angiostrongylus cantonensis is the most common infectious cause of eosinophilic meningitis. Timely diagnosis of these infections is difficult, partly because reliable laboratory diagnostic methods are unavailable. The aim of this study was to evaluate the usefulness of a real-time polymerase chain reaction (PCR) assay for the detection of A. cantonensis DNA in human cerebrospinal fluid (CSF) specimens. A total of 49 CSF specimens from 33 patients with eosinophilic meningitis were included: A. cantonensis DNA was detected in 32 CSF specimens, from 22 patients. Four patients had intermittently positive and negative real-time PCR results on subsequent samples, indicating that the level of A. cantonensis DNA present in CSF may fluctuate during the course of the illness. Immunodiagnosis and/or supplemental PCR testing supported the real-time PCR findings for 30 patients. On the basis of these observations, this real-time PCR assay can be useful to detect A. cantonensis in the CSF from patients with eosinophilic meningitis. © The American Society of Tropical Medicine and Hygiene.

  17. Near Re