Objective:Traditional methods for the detecction of C.jejuni are slow,labour-intensive and costly,especially when large numbers of samples are involved,In this report,we describe a PapManfluorogenic PCR system to rapidly detect and quantitate C.Jejuni.Methods: TapMan PCR uses a pair of primers and a fluorescently-labeled TaqMan probe,The C.jejuni-specific primers and probe were designed through a specific primer designing program based on C.jejuni-specific sequence,Campylobacter strains and non-campylobacters were tested with this assay.Spiked chicken rinse,milk and water samples were also tested.Results:This PCR assay detected all 22C.jejuni and two C.coli strains tested,while none of the 23 non-campylobacters tested positive.The assay was 100% specific for campylobacters,This assay can quantitate C.jejuni over a 5-log range,with the thershold of detection being less than 100 cfu in water and approximately 1000 cfu in chicken rines,The TaqMan PCR assay gave positive signals for chicken rinse,milk,and water samples inoculated with C.jejuni at 1.5-3.5cfu/25ml sample within 2h,after enrichment in Preston broth at 37℃ for 3-4h and then at 42℃ for 16-20h.Conclusion.:the Taqman PCR assay can be used for the detection and enumeration of C.jejuni from milk,chicken,and water samples with a short enrichment step.This technology should be useful for the food industry in monitoring levels of C.jejunit and C.coli at various critical control points in a food processing facility.
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