Topic
Multiplex polymerase chain reaction
About: Multiplex polymerase chain reaction is a research topic. Over the lifetime, 6409 publications have been published within this topic receiving 221244 citations.
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TL;DR: The specificity and the sensitivity of the ProFlu-1 assay ranged from 97% to 100% and the assay amplified 9% of samples not detected by conventional methods.
Abstract: We evaluated the one-step multiplex real-time reverse transcription-PCR ProFlu-1 assay for the detection of influenza A and influenza B viruses and respiratory syncytial viruses from 353 pediatric nasopharyngeal aspirates. As assessed by comparison with the results of immunofluorescence testing and cell culture, the specificity and the sensitivity of the ProFlu-1 assay ranged from 97% to 100%. In addition, the ProFlu-1 assay amplified 9% of samples not detected by conventional methods.
53 citations
Journal Article•
TL;DR: Multiplex PCR amplification with a pre-enrichment step was shown to be an effective, sensitive and rapid method for the simultaneous detection of these three microbiological parameters in drinking water.
Abstract: Multiplex PCR amplification of lacZ, uidA and plc genes was developed for the simulta- neous detection of total coliform bacteria for Escherichia coli and Clostridium perfringens, in drinking water. Detection by agarose gel electrophoresis yielded a band of 876 bp for the lacZ gene of all coliform bacteria; a band of 147 bp for the uidA gene and a band of 876 bp for the lacZ gene of all strains of E. coli; a band of 280 bp for the plc gene for all strains of C. perfringens; and a negative result for all three genes when tested with other bacteria. The detection limit was 100 pg for E. coli and C. perfringens, and 1 ng for coliform bacteria when measured with purified DNA. This assay was applied to the detection of these bacteria in spiked water samples. Spiked water samples with 0-1,000 CFU/ml of coliform bacteria and/or E. coli and/or C. perfringens were detected by this multiplex PCR after a pre-enrichment step to increase the sensitivity and to ensure that the detection was based on the presence of cultivable bacteria. The result of bacterial detection from the multiplex PCR was comparable with that of a standard plate count on selective medium (p=0.62). When using standard plate counts as a gold standard, the sensitivity for this test was 99.1 % (95% CI 95.33, 99.98) and the specificity was 90.9 % (95% CI 75.67, 98.08). Multiplex PCR amplifica- tion with a pre-enrichment step was shown to be an effective, sensitive and rapid method for the simultaneous detection of these three microbiological parameters in drinking water.
53 citations
TL;DR: The genetic diversity of B. cereus isolates was investigated using a random amplified polymorphic DNA (RAPD) assay and it was found that the discriminating ability of the OPG 16 primer was about threefold higher than that of MUP 3 in the RAPD assay.
52 citations
TL;DR: Limited field testing of this procedure demonstrated the utility of a ribosomal gene based species-specific malaria diagnosis by showing that the PCR procedure yields 1.4-kb and 0.5-kb DNA fragments for P. falciparum and P. vivax.
52 citations
TL;DR: Analysis of naturally contaminated water and food samples using agadAB-specific DNA probe that was labeled with digoxigenin showed that a gadAB-based assay is as reliable as standard methods that enumerate E. coli organisms on the basis of lactose fermentation.
Abstract: The enzyme glutamate decarboxylase (GAD) is prevalent in Escherichia coli but few strains in the various pathogenic E coli groups have been tested for GAD Using PCR primers that amplify a 670-bp segment from the gadA and gadB genes encoding GAD, we examined the distribution of the gadAB genes among enteric bacteria Analysis of 173 pathogenic E coli strains, including 125 enterohemorrhagic E coli isolates of the O157:H7 serotype and its phenotypic variants and 48 isolates of enteropathogenic E coli, enterotoxigenic E coli, enteroinvasive E coli, and other Shiga toxin-producing E coli (STEC) serotypes, showed that gadAB genes were present in all these strains Among the 22 non-E coli isolates tested, only the 6 Shigella spp carried gadAB Analysis of naturally contaminated water and food samples using a gadAB-specific DNA probe that was labeled with digoxigenin showed that a gadAB-based assay is as reliable as standard methods that enumerate E coli organisms on the basis of lactose fermentation The presence of few E coli cells initially seeded into produce rinsates could be detected by PCR to gadA/B genes after overnight enrichment A multiplex PCR assay using the gadAB primers in combination with primers to Shiga toxin (Stx) genes stx(1) and stx(2) was effective in detecting STEC from the enrichment medium after seeding produce rinsate samples with as few as 2 CFU The gadAB primers may be multiplexed with primers to other trait virulence markers to specifically identify other pathogenic E coli groups
52 citations