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: A highly specific real-time PCR strategy for the detection of Y. pestis was developed and evaluated, obtaining results within 3 h including DNA preparation, showing 100% accuracy when compared with combinations of conventional PCR assays.
Abstract: The objective of the present study was to establish a system of real-time polymerase chain reactions (PCRs) for the specific detection of Yersinia pestis using the LightCycler™ (LC) instrument. Twenty-five strains of Y. pestis, 94 strains of other Yersinia species and 33 clinically relevant bacteria were investigated. Assays for the 16S rRNA gene target and the plasminogen activator gene (resides on the 9.5-kb plasmid) and for the Y. pestis murine toxin gene and the fraction 1 antigen gene (both on the 100-kb plasmid) were combined for the use in two multiplex assays including an internal amplification control detecting bacteriophage λ-DNA. Applying these multiplex assays, Y. pestis was selectively identified; other bacteria yielded no amplification products. The lower limit of detection was approximately 0.1 genome equivalent. Rat or flea DNA had no inhibitory effects on the detection of Y. pestis. The results obtained using the multiplex real-time assays showed 100% accuracy when compared with combinations of conventional PCR assays. We developed and evaluated a highly specific real-time PCR strategy for the detection of Y. pestis, obtaining results within 3 h including DNA preparation.
83 citations
TL;DR: A real-time multiplex PCR was developed that identifies common diarrhea-causing bacteria in fecal samples and might be useful as an additional diagnostic tool whenever time is important in the diagnosis of enteropathogenic bacteria.
83 citations
TL;DR: Repeated sampling or cell culture in combination with multiplex PCR and with the incorporation of IL-2 in culture medium increased the sensitivity for detection of EHV in PBMC and demonstrated that EhV5 DNA could be identified inPBMC from 89% of foals and 100% of adult horses.
83 citations
TL;DR: Multiplex real-time PCR was found to be an equivalent or superior modality for the diagnosis of STIs and could be a cost-effective and rapid diagnostic tool for the simultaneous detection of multiple STI microorganisms.
83 citations
TL;DR: The multiplex PCR assay method was capable of detecting 5 colony-forming units of each of the three pathogens per 25 g of more than 40 types of food, and the detection rate of the PCR assay was higher than that of conventional culture methods.
Abstract: Conventional culture methods were compared to a multiplex polymerase chain reaction (PCR) assay for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 from enrichment cultures of various types of artificially inoculated and naturally contaminated foods. The multiplex PCR assay was evaluated in 44 types of spiked food samples, including meat, produce, fish, and dairy products targeting genes specific for each pathogen for simultaneous detection. The sensitivity of the assay was
82 citations