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Journal articles on the topic 'Quantitative real-time PCR TaqMan probe'

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Published: 4 June 2021

Last updated: 5 February 2022

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1

Lee, Jae Il, and In Seop Kim. "TaqMan Probe Real-Time PCR for Quantitative Detection of Mycoplasma during Manufacture of Biologics." KSBB Journal 29, no. 5 (2014): 361–71. http://dx.doi.org/10.7841/ksbbj.2014.29.5.361.

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Weller, S. A., J. G. Elphinstone, N. C. Smith, N. Boonham, and D. E. Stead. "Detection of Ralstonia solanacearumStrains with a Quantitative, Multiplex, Real-Time, Fluorogenic PCR (TaqMan) Assay." Applied and Environmental Microbiology 66, no. 7 (2000): 2853–58. http://dx.doi.org/10.1128/aem.66.7.2853-2858.2000.

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ABSTRACT A fluorogenic (TaqMan) PCR assay was developed to detectRalstonia solanacearum strains. Two fluorogenic probes were utilized in a multiplex reaction; one broad-range probe (RS) detected all biovars of R. solanacearum, and a second more specific probe (B2) detected only biovar 2A. Amplification of the target was measured by the 5′ nuclease activity of Taq DNA polymerase on each probe, resulting in emission of fluorescence. TaqMan PCR was performed with DNA extracted from 42 R. solanacearum and genetically or serologically related strains to demonstrate the specificity of the assay. In

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Jothikumar, Narayanan, Theresa L. Cromeans, Vincent R. Hill, Xiaoyan Lu, Mark D. Sobsey, and Dean D. Erdman. "Quantitative Real-Time PCR Assays for Detection of Human Adenoviruses and Identification of Serotypes 40 and 41." Applied and Environmental Microbiology 71, no. 6 (2005): 3131–36. http://dx.doi.org/10.1128/aem.71.6.3131-3136.2005.

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ABSTRACT A quantitative real-time TaqMan PCR assay for detection of human adenoviruses (HAdV) was developed using broadly reactive consensus primers and a TaqMan probe targeting a conserved region of the hexon gene. The TaqMan assay correctly identified 56 representative adenovirus prototype strains and field isolates from all six adenovirus species (A to F). Based on infectious units, the TaqMan assay was able to detect as few as 0.4 and 0.004 infectious units of adenovirus serotype 2 (AdV2) and AdV41, respectively, with results obtained in less than 90 min. Using genomic equivalents, the bro

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Liu, Ya-Mei, Liang Qiu, An-Zhi Sheng, Xiao-Yuan Wan, Dong-Yuan Cheng, and Jie Huang. "Quantitative detection method of Enterocytozoon hepatopenaei using TaqMan probe real-time PCR." Journal of Invertebrate Pathology 151 (January 2018): 191–96. http://dx.doi.org/10.1016/j.jip.2017.12.006.

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Rosenbergova, K., P. Lany, Z. Pospisil, O. Kubicek, V. Celer, and D. Molinkova. "Quantification of avian influenza virus in tissues of mute swans using TaqMan real time qRT-PCR." Veterinární Medicína 54, No. 9 (2009): 435–43. http://dx.doi.org/10.17221/16/2009-vetmed.

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This study reports on the first quantification of avian influenza virus in the organs of mute swans that died during the epizootic of avian influenza (H5N1) between January and April 2006 in the Czech Republic. The quantitative real-time Reverse Transcriptase PCR (qRT-PCR) assay based on a TaqMan probe was developed for a rapid detection and quantification of avian influenza virus RNA in clinical samples collected from mute swans. Conserved regions in the matrix protein gene of avian influenza virus served as targets for the primers and TaqMan probe design. A recombinant plasmid containing the

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Li, Jin, Lilin Wang, Pasi A. Jänne, and G. Mike Makrigiorgos. "Coamplification at Lower Denaturation Temperature-PCR Increases Mutation-Detection Selectivity of TaqMan-Based Real-Time PCR." Clinical Chemistry 55, no. 4 (2009): 748–56. http://dx.doi.org/10.1373/clinchem.2008.113381.

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Abstract Background: DNA genotyping with mutation-specific TaqMan® probes (Applied Biosystems) is broadly used in detection of single-nucleotide polymorphisms but is less so for somatic mutations because of its limited selectivity for low-level mutations. We recently described coamplification at lower denaturation temperature-PCR (COLD-PCR), a method that amplifies minority alleles selectively from mixtures of wild-type and mutation-containing sequences during the PCR. We demonstrate that combining COLD-PCR with TaqMan technology provides TaqMan genotyping with the selectivity needed to detect

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Jiang, Qianli, Shan Jiang, Fanyi Meng, et al. "Efficient Duplex Real-Time Quantitative RT-PCR of BCR-ABL and ABL Gene Transcripts for Monitoring of Minimal Residual Disease." Blood 112, no. 11 (2008): 4887. http://dx.doi.org/10.1182/blood.v112.11.4887.4887.

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Abstract BCL-ABL fusion gene can be found in 100% chronic myelogenous leukemia (CML) and about 25% adult acute lympholid leukemia where its expression level is a crucial parameter for monitoring of minimal residual disease (MRD). Depending on the breakpoint in BCR, exon 2 of ABL (a2) joins with exons 1 (e1), 13 (b2), or 14 (b3), or rarely to exon 19 (e19) of BCR resulting in chimeric proteins of p190, p210 and p230, respectively. The c-ABL gene is one of the best controls for MRD detection by real-time quantitative RT-PCR (RQ-RT-PCR). In most studies published before, PCR probes were labeled b

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Pusterla, Nicola, Jon B. Huder, Christian M. Leutenegger, Ueli Braun, John E. Madigan, and Hans Lutz. "Quantitative Real-Time PCR for Detection of Members of the Ehrlichia phagocytophila Genogroup in Host Animals and Ixodes ricinus Ticks." Journal of Clinical Microbiology 37, no. 5 (1999): 1329–31. http://dx.doi.org/10.1128/jcm.37.5.1329-1331.1999.

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A TaqMan PCR was established for identification and quantitation of members of the Ehrlichia phagocytophila group in experimentally infected cows and in Ixodes ricinus ticks. The TaqMan PCR identified a 106-bp section of the 16S rRNA gene by use of a specific fluorogenic probe and two primers. This technique was specific for members of the E. phagocytophila group, which include E. phagocytophila, Ehrlichia equi, and the agent of human granulocytic ehrlichiosis. The TaqMan system identified 10 copies of a cloned section of the 16S rRNA gene ofE. phagocytophila. The sensitivity and specificity o

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Boyer, S., and J. Yang. "Measurement of gene amplification in FFPE tumor tissues by quantitative real-time PCR in comparison with FISH." Journal of Clinical Oncology 25, no. 18_suppl (2007): 14086. http://dx.doi.org/10.1200/jco.2007.25.18_suppl.14086.

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14086 Background: Amplified oncogenes such as HER2 and EGFR have proven to be good targets for molecular therapy. Recent retrospective analyses suggest that EGFR amplification predicate better responses to EGFR kinase inhibitors. However, different methods were used to measure EGFR gene amplification and polysomy in these studies, including FISH and various real-time quantitative PCR platforms, which may have led to the different conclusions reported by these groups. In this study, we compared Taqman- QPCR with FISH in scoring EGFR and HER2 amplification and EGFR polysomy on 42 FFPE tumor samp

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10

Lin, Mei-Hui, Tse-Ching Chen, Tseng-tong Kuo, Ching-Chung Tseng, and Ching-Ping Tseng. "Real-Time PCR for Quantitative Detection ofToxoplasma gondii." Journal of Clinical Microbiology 38, no. 11 (2000): 4121–25. http://dx.doi.org/10.1128/jcm.38.11.4121-4125.2000.

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The protozoan Toxoplasma gondii is one of the most common infectious pathogenic parasites and can cause severe medical complications in infants and immunocompromised individuals. We report here the development of a real-time PCR-based assay for the detection of T. gondii. Oligonucleotide primers and a fluorescence-labeled TaqMan probe were designed to amplify the T. gondii B1 gene. After 40 PCR cycles, the cycle threshold values (CT) indicative of the quantity of the target gene were determined. Typically, a CT of 25.09 was obtained with DNA from 500 tachyzoites of the T. gondii RH strain. The

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Winton, L. M., J. K. Stone, L. S. Watrud, and E. M. Hansen. "Simultaneous One-Tube Quantification of Host and Pathogen DNA with Real-Time Polymerase Chain Reaction." Phytopathology® 92, no. 1 (2002): 112–16. http://dx.doi.org/10.1094/phyto.2002.92.1.112.

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Phaeocryptopus gaeumannii is a widespread foliar parasite of Douglas-fir. Although normally innocuous, the fungus also causes the defoliating disease Swiss needle cast in heavily infected needles. The extent of P. gaeumannii colonization in Douglas-fir foliage was estimated with real-time quantitative polymerase chain reaction (PCR) using TaqMan chemistry. In order to derive a normalized expression of colonization, both pathogen and host DNA were simultaneously amplified but individually detected by species-specific primers and TaqMan probes labeled with different fluorescent dyes. Detection o

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Hein, Ingeborg, Angelika Lehner, Petra Rieck, Kurt Klein, Ernst Brandl, and Martin Wagner. "Comparison of Different Approaches To QuantifyStaphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese." Applied and Environmental Microbiology 67, no. 7 (2001): 3122–26. http://dx.doi.org/10.1128/aem.67.7.3122-3126.2001.

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ABSTRACT Two different real-time quantitative PCR (RTQ-PCR) approaches were applied for PCR-based quantification of Staphylococcus aureus cells by targeting the thermonuclease (nuc) gene. Purified DNA extracts from pure cultures ofS. aureus were quantified in a LightCycler system using SYBR Green I. Quantification proved to be less sensitive (60nuc gene copies/μl) than using a fluorigenic TaqMan probe (6 nuc gene copies/μl). Comparison of the LightCycler system and the well-established ABI Prism 7700 SDS with TaqMan probes revealed no statistically significant differences with respect to sensi

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Mingxiao, Ma, Liu Jinhua, Song Yingjin, Li Li, and Li Yongfei. "TaqMan MGB Probe Fluorescence Real-Time Quantitative PCR for Rapid Detection of Chinese Sacbrood Virus." PLoS ONE 8, no. 2 (2013): e52670. http://dx.doi.org/10.1371/journal.pone.0052670.

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14

PEGELS, NICOLETTE, ISABEL GONZÁLEZ, TERESA GARCÍA, and ROSARIO MARTÍN. "Detection of Banned Ruminant-Derived Material in Industrial Feedstuffs by TaqMan Real-Time PCR Assay." Journal of Food Protection 74, no. 8 (2011): 1300–1308. http://dx.doi.org/10.4315/0362-028x.jfp-11-029.

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A ruminant-specific real-time PCR system was designed and applied for the detection of processed animal protein from ruminants in industrial feedstuffs. The assay includes a primer pair and a TaqMan probe selectively targeting mitochondrial 16S rRNA gene sequences from the ruminant group and another primer-probe set based on the eukaryotic nuclear 18S rRNA gene (positive amplification control). Both ruminant and eukaryotic PCR systems generated short PCR amplicons of 79 and 77 bp, respectively. To evaluate the suitability of the real-time PCR assay for the detection of banned by-products of ru

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15

Nakhla, Mark K., Kristina J. Owens, Wenbin Li, Gang Wei, Andrea M. Skantar, and Laurene Levy. "Multiplex Real-Time PCR Assays for the Identification of the Potato Cyst and Tobacco Cyst Nematodes." Plant Disease 94, no. 8 (2010): 959–65. http://dx.doi.org/10.1094/pdis-94-8-0959.

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TaqMan primer-probe sets were developed for the detection and identification of potato cyst nematodes (PCNs) Globodera pallida and G. rostochiensis using two-tube, multiplex real-time polymerase chain reaction (PCR). One tube contained a primer-probe set specific for G. pallida (pale potato cyst nematode) multiplexed with another primer-probe set specific for G. rostochiensis (golden potato cyst nematode). A second tube consisted of the G. pallida-specific primer-probe set multiplexed with a primer-probe set specific for G. tabacum (the morphologically similar tobacco cyst nematode). This inte

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16

Ko, Woon Young, Na Gyeong Noh, and In Seop Kim. "TaqMan probe real-time PCR for quantitative detection of bovine adenovirus type 1 during the manufacture of biologics and medical devices using bovine-derived raw materials." Korean Journal of Microbiology 51, no. 3 (2015): 199–208. http://dx.doi.org/10.7845/kjm.2015.5036.

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17

Kirchner, Sebastian, K. Melanie Krämer, Martin Schulze, et al. "Pentaplexed Quantitative Real-Time PCR Assay for the Simultaneous Detection and Quantification of Botulinum Neurotoxin-Producing Clostridia in Food and Clinical Samples." Applied and Environmental Microbiology 76, no. 13 (2010): 4387–95. http://dx.doi.org/10.1128/aem.02490-09.

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ABSTRACT Botulinum neurotoxins are produced by the anaerobic bacterium Clostridium botulinum and are divided into seven distinct serotypes (A to G) known to cause botulism in animals and humans. In this study, a multiplexed quantitative real-time PCR assay for the simultaneous detection of the human pathogenic C. botulinum serotypes A, B, E, and F was developed. Based on the TaqMan chemistry, we used five individual primer-probe sets within one PCR, combining both minor groove binder- and locked nucleic acid-containing probes. Each hydrolysis probe was individually labeled with distinguishable

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Jabłoński, Artur, Dominika Borowska, Sylwia Zębek, et al. "Application of quantitative PCR for detection of Mycoplasma suis in blood samples." Bulletin of the Veterinary Institute in Pulawy 58, no. 4 (2014): 533–39. http://dx.doi.org/10.2478/bvip-2014-0082.

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Abstract The aim of the study was to develop and validate a real-time PCR method, using a TaqMan probe, for quantification of Mycoplasma suis in porcine blood. No PCR signals with closely related non-haemotrophic mycoplasmas were obtained. The detection limit of PCR for plasmid combined with blood DNA was determined to be 103/reaction (5 μL of DNA) (1.2x105 target copies in 1 mL of blood). The linearity of real-time PCR (near 1) indicates its use as a quantitative method. Real-time and quantitative PCR were sensitive and specific for the detection and quantification of M. suis in the blood of

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Becker, Sven, Peter Böger, Ralfh Oehlmann, and Anneliese Ernst. "PCR Bias in Ecological Analysis: a Case Study for Quantitative Taq Nuclease Assays in Analyses of Microbial Communities." Applied and Environmental Microbiology 66, no. 11 (2000): 4945–53. http://dx.doi.org/10.1128/aem.66.11.4945-4953.2000.

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ABSTRACT Succession of ecotypes, physiologically diverse strains with negligible rRNA sequence divergence, may explain the dominance of small, red-pigmented (phycoerythrin-rich) cyanobacteria in the autotrophic picoplankton of deep lakes (C. Postius and A. Ernst, Arch. Microbiol. 172:69–75, 1999). In order to test this hypothesis, it is necessary to determine the abundance of specific ecotypes or genotypes in a mixed background of phylogenetically similar organisms. In this study, we examined the performance of Taq nuclease assays (TNAs), PCR-based assays in which the amount of an amplicon is

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20

Fu, Hua-Ying, Sheng-Ren Sun, Jin-Da Wang, et al. "Rapid and Quantitative Detection ofLeifsonia xylisubsp.xyliin Sugarcane Stalk Juice Using a Real-Time Fluorescent (TaqMan) PCR Assay." BioMed Research International 2016 (2016): 1–8. http://dx.doi.org/10.1155/2016/2681816.

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Ratoon stunting disease (RSD) of sugarcane, one of the most important diseases seriously affecting the productivity of sugarcane crops, was caused by the bacterial agentLeifsonia xylisubsp.xyli(Lxx). A TaqMan probe-based real-time quantitative polymerase chain reaction (qPCR) assay was established in this study for the quantification ofLxxdetection in sugarcane stalk juice. A pair of PCR primers (Pat1-QF/Pat1-QR) and a fluorogenic probe (Pat1-QP) targeting thePart1gene ofLxxwere used for the qPCR assay. The assay had a detection limit of 100 copies of plasmid DNA and 100 fg ofLxxgenomic DNA, w

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Schmidt, Anna-mary, and Michael E. Rott. "Real-Time Polymerase Chain Reaction (PCR) Quantitative Detection ofBrassica napusUsing a Locked Nucleic Acid TaqMan Probe." Journal of Agricultural and Food Chemistry 54, no. 4 (2006): 1158–65. http://dx.doi.org/10.1021/jf052036m.

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22

AMOAKO, KINGSLEY K., NORIKO GOJI, TREVOR MACMILLAN, et al. "Development of Multitarget Real-Time PCR for the Rapid, Specific, and Sensitive Detection of Yersinia pestis in Milk and Ground Beef." Journal of Food Protection 73, no. 1 (2010): 18–25. http://dx.doi.org/10.4315/0362-028x-73.1.18.

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Real-time PCR has been used previously to detect Yersinia pestis; this study applies this rapid, specific, and sensitive nucleic acid–based method to the detection and quantitation of Y. pestis specifically in food. Five sets of primers and corresponding TaqMan dual-labelled fluorogenic hybridization probes for Y. pestis were designed and optimized for specificity testing using genomic DNA from 71 bacterial strains. Four Y. pestis–specific primer and probe sets were developed, based on the virulence plasmid targets, and used to distinguish this bacterium from the various Yersinia and other bac

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23

Cheng, Ching-Yang, Jing-Ruei Chi, Sin-Rong Lin, Chi-Chiang Chou, and Chin-Cheng Huang. "Rapid quantification of Salmonella Typhimurium inoculated to meat products by real-time PCR." Acta Veterinaria Hungarica 57, no. 1 (2009): 25–38. http://dx.doi.org/10.1556/avet.57.2009.1.3.

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The objective of this study was to use a 5′-nuclease (TaqMan) real-time PCR method with primers and probe specific to thespaQgene as a rapid approach to quantitatively determineSalmonellaTyphimurium. The result showed that the correlation coefficient between real-time PCR estimates and bovine serum albumin (BSA) plate counts ofS. Typhimurium was 0.99, independently of 105-fold numbers of bystanderEscherichia coliO157:H7 or total viable counts. The sensitivity of the real-time quantitative PCR assay was 10 CFU/mL for pureS. Typhimurium culture without enrichment. A known number ofS. Typhimurium

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Bai, Lijun, Yi-Mo Deng, Anthony J. Dodds, Sam Milliken, John Moore, and David D. F. Ma. "A SYBR Green - Based Real Time PCR Method for Detection of Haemopoietic Chimerism in Allogeneic Stem Cell Transplant Recipients." Blood 106, no. 11 (2005): 5222. http://dx.doi.org/10.1182/blood.v106.11.5222.5222.

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Abstract Aim Detection of donor cells (chimerism) in patients post allogeneic stem cell transplantation is for monitoring engraftment, early detection of graft rejection and disease relapse. Current methods include cytogenetics, blood grouping and DNA microsatellite test. Our aim is to develop a reliable and rapid real-time quantitative PCR (Q-PCR) method using SYBR green for detecting chimerism in allogeneic haemopoietic stem cell transplant recipients. Methods Twelve specific nucleotide polymorphisms (NPs) on 11 different chromosomes (including X, Y) were selected. Specific primers and fluor

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Hall, S. J., P. Hugenholtz, N. Siyambalapitiya, J. Keller, and L. L. Blackall. "The development and use of real-time PCR for the quantification of nitrifiers in activated sludge." Water Science and Technology 46, no. 1-2 (2002): 267–72. http://dx.doi.org/10.2166/wst.2002.0488.

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Chemical analytical data has long been used to monitor the performance of activated sludge plants even though the process relies on the performance of microorganisms. It is now evident that a rapid and reliable quantitative method is required, to be able to monitor the organisms responsible for nutrient transformation and their activities, allowing avenues for more efficient nutrient removal. The development of real-time or quantitative polymerase chain reaction (PCR) also known as TaqMan® or 5′-nuclease assay has allowed the rapid, quantitative analysis of DNA templates, eliminating some of t

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Yue, Zhi-Qin, Hong Liu, Wei-Ji Wang, Zhi-Wen Lei, Cheng-Zhu Liang, and Yu-Lin Jiang. "Development of Real-Time Polymerase Chain Reaction Assay with TaqMan Probe for the Quantitative Detection of Infectious Hypodermal and Hematopoietic Necrosis Virus from Shrimp." Journal of AOAC INTERNATIONAL 89, no. 1 (2006): 240–44. http://dx.doi.org/10.1093/jaoac/89.1.240.

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Abstract An assay was developed for the detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV) based on real-time quantitative polymerase chain reaction (PCR). A pair of primers and a TaqMan probe were designed that are specific for the recognition of a conservative region in the IHHNV genome. The IHHNV real-time PCR assay had a detection limit of 9 DNA copies,with a dynamic range of detection between 9 106 and 9 DNA copies. The primer pairs and probe were specific to IHHNV and did not cross-reactwith shrimp genomic DNAor other shrimp viruses such as White Spot Syndrome Vi

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Steinberg, Lisa M., and John M. Regan. "mcrA-Targeted Real-Time Quantitative PCR Method To Examine Methanogen Communities." Applied and Environmental Microbiology 75, no. 13 (2009): 4435–42. http://dx.doi.org/10.1128/aem.02858-08.

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ABSTRACT Methanogens are of great importance in carbon cycling and alternative energy production, but quantitation with culture-based methods is time-consuming and biased against methanogen groups that are difficult to cultivate in a laboratory. For these reasons, methanogens are typically studied through culture-independent molecular techniques. We developed a SYBR green I quantitative PCR (qPCR) assay to quantify total numbers of methyl coenzyme M reductase α-subunit (mcrA) genes. TaqMan probes were also designed to target nine different phylogenetic groups of methanogens in qPCR assays. Tot

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Martï¿½n, Belï¿½n, Anna Jofrï¿½, Margarita Garriga, Maria Pla, and Teresa Aymerich. "Rapid Quantitative Detection of Lactobacillus sakei in Meat and Fermented Sausages by Real-Time PCR." Applied and Environmental Microbiology 72, no. 9 (2006): 6040–48. http://dx.doi.org/10.1128/aem.02852-05.

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ABSTRACT 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 qua

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Hu, Chenyan, Zhongzhu Yang, Zhen Song, Linghui Xiao, and Yang He. "A strategy for preparing non-fluorescent graphene oxide quantum dots as fluorescence quenchers in quantitative real-time PCR." RSC Advances 10, no. 25 (2020): 14944–52. http://dx.doi.org/10.1039/d0ra00142b.

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Ruiz-Ruiz, S., P. Moreno, J. Guerri, and S. Ambrós. "Discrimination Between Mild and Severe Citrus tristeza virus Isolates with a Rapid and Highly Specific Real-Time Reverse Transcription-Polymerase Chain Reaction Method Using TaqMan LNA Probes." Phytopathology® 99, no. 3 (2009): 307–15. http://dx.doi.org/10.1094/phyto-99-3-0307.

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Severe isolates of Citrus tristeza virus (CTV) inducing seedling yellows (SY) and/or stem pitting (SP) in grapefruit or sweet orange are a major threat for the citrus industry worldwide. Identification of these CTV variants was achieved by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) using a general primer set and three TaqMan locked nucleic acids (LNA) probes targeting sequences characteristic of severe, mild (non-SY, non-SP), and T36-like isolates. Successful amplification was achieved from fresh or silica-desiccated CTV-infected samples and all isolates bu

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Su, Yachun, Shanshan Wang, Jinlong Guo, Bantong Xue, Liping Xu, and Youxiong Que. "A TaqMan Real-Time PCR Assay for Detection and Quantification ofSporisorium scitamineumin Sugarcane." Scientific World Journal 2013 (2013): 1–9. http://dx.doi.org/10.1155/2013/942682.

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Sporisorium scitamineumis a fungal smut pathogen epidemic in sugarcane producing areas. Early detection and proper identification of the smut are an essential requirement in its management practice. In this study, we developed a TaqMan real-time PCR assay using specific primers (bEQ-F/bEQ-R) and a TaqMan probe (bEQ-P) which were designed based on thebE(b East mating type) gene (Genbank Accession no. U61290.1). This method was more sensitive (a detection limit of 10 ag pbE DNA and 0.8 ng sugarcane genomic DNA) than that of conventional PCR (10 fg and 100 ng, resp.). Reliability was demonstrated

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WANG, LUXIN, YONG LI, and AZLIN MUSTAPHA. "Rapid and Simultaneous Quantitation of Escherichia coli O157:H7, Salmonella, and Shigella in Ground Beef by Multiplex Real-Time PCR and Immunomagnetic Separation†." Journal of Food Protection 70, no. 6 (2007): 1366–72. http://dx.doi.org/10.4315/0362-028x-70.6.1366.

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The objective of this study was to establish a multiplex real-time PCR for the simultaneous quantitation of Escherichia coli O157:H7, Salmonella, and Shigella. Genomic DNA for the real-time PCR was extracted by the boiling method. Three sets of primers and corresponding TaqMan probes were designed to target these three pathogenic bacteria. Multiplex real-time PCR was performed with TaqMan Universal PCR Master Mix in an ABI Prism 7700 Sequence Detection System. Final standard curves were calculated for each pathogen by plotting the threshold cycle value against the bacterial number (log CFU per

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Liang, D. W., T. Zhang, and H. H. P. Fang. "Real-time quantifications of dominant anaerobes in an upflow reactor by polymerase chain reaction using a TaqMan probe." Water Science and Technology 57, no. 11 (2008): 1851–55. http://dx.doi.org/10.2166/wst.2008.042.

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This study was conducted to demonstrate the application of quantitative real-time polymerase chain reaction (qRT-PCR) for the quantification of dominant bacteria in an anaerobic reactor using a designed TaqMan probe. A novel group of uncultured thermophilic bacteria affiliated with Thermotogales was first found in a phenol-degrading sludge from a 55°C upflow anaerobic sludge blanket (UASB) reactor, which effectively removed 99% of phenol at loading of 0.51 g-phenol l−1 d−1 h of hydraulic retention. A TaqMan probe was then designed targeting this group of Thermotogales affiliated bacteria (TAB)

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BELLAGAMBA, FEDERICA, SERGIO COMINCINI, LUCA FERRETTI, FRANCO VALFRÈ, and VITTORIO M. MORETTI. "Application of Quantitative Real-Time PCR in the Detection of Prion-Protein Gene Species-Specific DNA Sequences in Animal Meals and Feedstuffs." Journal of Food Protection 69, no. 4 (2006): 891–96. http://dx.doi.org/10.4315/0362-028x-69.4.891.

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This study describes a method for quantitative and species-specific detection of animal DNA from different species (cattle, sheep, goat, swine, and chicken) in animal feed and feed ingredients, including fish meals. A quantitative real-time PCR approach was carried out to characterize species-specific sequences based on the amplification of prion-protein sequence. Prion-protein species-specific primers and TaqMan probes were designed, and amplification protocols were optimized in order to discriminate the different species with short PCR amplicons. The real-time quantitative PCR approach was a

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Cubero, J., and J. H. Graham. "Quantitative Real-Time Polymerase Chain Reaction for Bacterial Enumeration and Allelic Discrimination to Differentiate Xanthomonas Strains on Citrus." Phytopathology® 95, no. 11 (2005): 1333–40. http://dx.doi.org/10.1094/phyto-95-1333.

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Quantitative real-time polymerase chain reaction (QRT-PCR) was developed for identification and enumeration of bacteria in citrus plant samples infected with Xanthomonas axonopodis pvs. citri and citrumelo, the cause of citrus bacterial canker (CBC) and citrus bacterial spot (CBS), respectively. Three sets of primers based on the pathogenicity gene (pth) in X. axonopodis pv. citri, a ribosomal gene in X. axonopodis pv. citrumelo, and the leucine-responsive regulatory protein (lrp) in both pathovars were combined with TaqMan probes and applied for specific strain detection and quantification. C

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Fu, Wei-Lin, Sheng-Ren Sun, Hua-Ying Fu, Ru-Kai Chen, Jin-Wei Su, and San-Ji Gao. "A One-Step Real-Time RT-PCR Assay for the Detection and Quantitation ofSugarcane Streak Mosaic Virus." BioMed Research International 2015 (2015): 1–9. http://dx.doi.org/10.1155/2015/569131.

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Sugarcane mosaic disease is caused by theSugarcane streak mosaic virus(SCSMV; genusPoacevirus, familyPotyviridae) which is common in some Asian countries. Here, we established a protocol of a one-step real-time quantitative reverse transcription PCR (real-time qRT-PCR) using the TaqMan probe for the detection of SCSMV in sugarcane. Primers and probes were designed within the conserved region of the SCSMV coat protein (CP) gene sequences. Standard single-stranded RNA (ssRNA) generated by PCR-based gene transcripts of recombinant pGEM-CP plasmidin vitroand total RNA extracted from SCSMV-infected

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Dors, Arkadiusz, Andrzej Kowalczyk, and Małgorzata Pomorska-Mól. "Real-time quantitative PCR for detection and identification of Actinobacillus pleuropneumoniae serotype 2." Journal of Veterinary Research 60, no. 3 (2016): 253–56. http://dx.doi.org/10.1515/jvetres-2016-0038.

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AbstractIntroduction: Porcine pleuropneumonia inflicts important economic losses on most commercial herds. Detection of subclinical or chronic infection in animals still remains a challenge, as isolation and identification of A. pleuropneumoniae serotypes is difficult and quantification of the bacteria on agar plates is often almost impossible. The aim of the study was to develop and evaluate a serotype-specific quantitative TaqMan probe-based PCR for detection of serotype 2 in pig lungs, tonsils, and nasal swabs.Material and Methods: The primers were designed from the capsular polysaccharide

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SHARMA, VIJAY K. "Detection and Quantitation of Enterohemorrhagic Escherichia coli O157, O111, and O26 in Beef and Bovine Feces by Real-Time Polymerase Chain Reaction†." Journal of Food Protection 65, no. 9 (2002): 1371–80. http://dx.doi.org/10.4315/0362-028x-65.9.1371.

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Enterohemorrhagic Escherichia coli (EHEC) O157:H7 and certain non-O157 EHEC serotypes (such as O26:H11, O26: NM, O111:H8, and O111:NM) have emerged as significant causes of human disease throughout the world. Important virulence attributes of EHEC are the intimin protein (encoded by the eae gene) and Shiga toxins 1 and 2 (encoded by the stx1 and stx2 genes, respectively). Two sets of real-time polymerase chain reaction (R-PCR) assays were developed for the simultaneous detection and quantitation of EHEC through the monitoring of the presence of the eae and stx genes, and these assays were eval

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Bælum, Jacob, and Carsten S. Jacobsen. "TaqMan Probe-Based Real-Time PCR Assay for Detection and Discrimination of Class I, II, and III tfdA Genes in Soils Treated with Phenoxy Acid Herbicides." Applied and Environmental Microbiology 75, no. 9 (2009): 2969–72. http://dx.doi.org/10.1128/aem.02051-08.

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ABSTRACT Separate quantification of three classes of tfdA genes was performed using TaqMan quantitative real-time PCR for 13 different soils subsequent to mineralization of three phenoxy acids. Class III tfdA genes were found to be involved in mineralization more often than class I and II tfdA genes.

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Liu, Chun, Yanming M. Guo, Jizhen Z. Cao, et al. "Detection and quantification of Aeromonas schubertii in Channa maculata by TaqMan MGB probe fluorescence real‐time quantitative PCR." Journal of Fish Diseases 42, no. 1 (2018): 109–17. http://dx.doi.org/10.1111/jfd.12911.

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Coolong, Timothy W., Ronald R. Walcott, and William M. Randle. "A Quantitative Real-time Polymerase Chain Reaction Assay for Botrytis aclada in Onion Bulb Tissue." HortScience 43, no. 2 (2008): 408–13. http://dx.doi.org/10.21273/hortsci.43.2.408.

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A real-time polymerase chain reaction (PCR) assay has been developed for the detection and quantification of Botrytis aclada (Fresenius), a causal agent of neck rot in onion (Allium cepa L.) bulbs. The assay uses TaqMan probe-based chemistry to detect an amplicon from the L45-550 region of B. aclada while using a DNA sequence from the onion serine acetyl transferase gene (SAT1) as a control. The assay detection limits for B. aclada and onion were 10 pg·μL−1 of genomic DNA. The detection limit for lyophilized B. aclada mycelium was 1 μg. The presence of onion tissue in the samples did not affec

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KRCMAR, PAVEL, and EVA RENCOVA. "Quantitative Detection of Species-Specific DNA in Feedstuffs and Fish Meals." Journal of Food Protection 68, no. 6 (2005): 1217–21. http://dx.doi.org/10.4315/0362-028x-68.6.1217.

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A sensitive and rapid method for the quantitative detection of bovine-, ovine-, swine-, and chicken-specific mitochondrial DNA sequences based on real-time PCR has been developed. The specificity of the primers and probes for real-time PCR has been tested using DNA samples of other vertebrate species that may also be present in rendered products. The quantitative detection was performed with dual-labeled probes (TaqMan) using absolute quantification with external standards of single species meat-and-bone meals. This method facilitates the detection of 0.01% of the target species–derived materi

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Labuhn, M., V. Vuaroqueaux, F. Fina, et al. "Simultaneous Quantitative Detection of Relevant Biomarkers in Breast Cancer by Quantitative Real-Time PCR." International Journal of Biological Markers 21, no. 1 (2006): 30–39. http://dx.doi.org/10.1177/172460080602100105.

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The assessment of ERα, PgR and HER2 status is routinely performed today to determine the endocrine responsiveness of breast cancer samples. Such determination is usually accomplished by means of immunohistochemistry and in case of HER2 amplification by means of fluorescent in situ hybridization (FISH). The analysis of these markers can be improved by simultaneous measurements using quantitative real-time PCR (Qrt-PCR). In this study we compared Qrt-PCR results for the assessment of mRNA levels of ERα, PgR, and the members of the human epidermal growth factor receptor family, HER1, HER2, HER3 a

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Hou, Yi-Xuan, Chun Xie, Kang Wang, et al. "Development and application of a TaqMan-MGB real-time RT-PCR assay for the detection of porcine epidemic diarrhoea virus strains in China." Journal of Veterinary Research 60, no. 2 (2016): 127–33. http://dx.doi.org/10.1515/jvetres-2016-0018.

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AbstractIntroduction:A real-time RT-PCR method for identification and quantification of porcine epidemic diarrhoea virus (PEDV) strains in China was developed.Material and Methods:Based on the conserved sequence of the PEDV nucleocapsid (N) gene, a primer pair and probe were designed to establish a TaqMan-MGB real-time RT-PCR assay for quantitative detection of the virus. The sequence was cloned into the pMD18-T vector and a series of diluted recombinant plasmids were used to generate a standard curve with an R2 value of 0.999.Results:The developed quantitative PCR assay detected viral titres

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Alizadeh, Mehdi, Marc Bernard, Bruno Danic, et al. "Quantitative assessment of hematopoietic chimerism after bone marrow transplantation by real-time quantitative polymerase chain reaction." Blood 99, no. 12 (2002): 4618–25. http://dx.doi.org/10.1182/blood.v99.12.4618.

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We have developed a real-time quantitative polymerase chain reaction (PCR) assay using TaqMan technology (Applied Biosystems, Foster City, CA) for monitoring donor cell engraftment in allogenic hematopoietic stem cell transplant recipients. For this purpose, we selected 19 specific sequence polymorphisms belonging to 11 human biallelic loci located on 9 different chromosomes. Using a set of specially designed primers and fluorogenic probes, we evaluated the 19 markers' informativity on a panel of 126 DNA samples from 63 recipient/donor pairs. In more than 90% of these pairs, discrimination bet

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Chabou, S., T. Leangapichart, L. Okdah, S. Le Page, L. Hadjadj, and J. M. Rolain. "Real-time quantitative PCR assay with Taqman ® probe for rapid detection of MCR-1 plasmid-mediated colistin resistance." New Microbes and New Infections 13 (September 2016): 71–74. http://dx.doi.org/10.1016/j.nmni.2016.06.017.

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Crosslin, J. M., G. J. Vandemark, and J. E. Munyaneza. "Development of a Real-Time, Quantitative PCR for Detection of the Columbia Basin Potato Purple Top Phytoplasma in Plants and Beet Leafhoppers." Plant Disease 90, no. 5 (2006): 663–67. http://dx.doi.org/10.1094/pd-90-0663.

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A quantitative, real-time “TaqMan” polymerase chain reaction assay (real-time PCR) was developed which was capable of detecting and quantifying a group 16SrVI phytoplasma in DNA extracts prepared from infected tomatoes, potatoes, and beet leafhoppers (Circulifer tenellus). Primers and probe were designed from the 16S rRNA gene of the Columbia Basin potato purple top phytoplasma, which is closely related to the beet leafhopper transmitted virescence agent. The detection limit in phytoplasma-infected tomato DNA was approximately 50 pg. The concentration of phytoplasma varied considerably among p

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Rodríguez-Lázaro, David, Maria Pla, Mariela Scortti, Héctor J. Monzó, and José A. Vázquez-Boland. "A Novel Real-Time PCR for Listeria monocytogenes That Monitors Analytical Performance via an Internal Amplification Control." Applied and Environmental Microbiology 71, no. 12 (2005): 9008–12. http://dx.doi.org/10.1128/aem.71.12.9008-9012.2005.

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ABSTRACT We describe a novel quantitative real-time (Q)-PCR assay for Listeria monocytogenes based on the coamplification of a target hly gene fragment and an internal amplification control (IAC). The IAC is a chimeric double-stranded DNA containing a fragment of the rapeseed BnACCg8 gene flanked by the hly-specific target sequences. This IAC is detected using a second TaqMan probe labeled with a different fluorophore, enabling the simultaneous monitoring of the hly and IAC signals. The hly-IAC assay had a specificity and sensitivity of 100%, as assessed using 49 L. monocytogenes isolates of d

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Van der Heyden, Hervé, Thérèse Wallon, C. André Lévesque, and Odile Carisse. "Detection and Quantification of Pythium tracheiphilum in Soil by Multiplex Real-Time qPCR." Plant Disease 103, no. 3 (2019): 475–83. http://dx.doi.org/10.1094/pdis-03-18-0419-re.

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In Canada, head lettuce (Lactuca sativa capitata) is extensively produced in the muck soils of southwestern Québec. However, yields are increasingly affected by various soilborne pathogens, including Pythium spp., which cause wilt and damping off. In a survey conducted in Québec muck soils in 2010 and 2011, Pythium tracheiphilum Matta was identified as the predominant Pythium sp. in the root of head lettuce showing Pythium stunt symptoms. Therefore, to improve risk assessment and help further understanding of disease epidemiology, a specific and sensitive real-time quantitative polymerase chai

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Natonek-Wiśniewska, Małgorzata, and Piotr Krzyścin. "The use of PCR and Real-Time PCR for Qualitative and Quantitative Determination of Poultry and Chicken Meals." Annals of Animal Science 16, no. 3 (2016): 731–41. http://dx.doi.org/10.1515/aoas-2016-0003.

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AbstractEuropean Union regulations (e.g. Commission Regulation (EU) No 56/2013) set restrictions on the use of poultry meals in animal nutrition, which requires the species composition of manufactured feeds to be constantly monitored. The aim of this study is to develop a method for qualitative analysis of poultry meals, enabling the four main poultry species (chicken, turkey, duck, goose) to be detected using one pair of primers, and a method for quantitative determination of chicken meals in poultry meals. The qualitative identification method was developed using PCR technology, whereas qPCR

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