Development and application of multiplex real-time PCR for detection of pathogenic bacteria in aquatic products
Abstract：A multiplex real-time PCR method was developed for the simultaneous detection of Salmonella spp, Vibrio parahaemolytichus and Escherichia Coli O157：H7 based on the primers and probes designed based on the conserved domains of invA gene for Salmonella spp, toxR gene for Vibrio parahaemolytichus and RFBE gene for Escherichia Coli O157： }t7- The developed multiplex real-time PCR method could detect less than 10efu per PCR reaction for all the pathogenic bacteria above in pure cultured broth and less than 10cfu per 25g for 6h-cultured artificially inoculated samples. Only target strains of 48 reference/standard strains were detected positive in the specificity testing. The intra- and inter-assay coefficients of variation were less than 2%. The method was applied to samples artificially coniaminated with these pathogenic bacteria and samples collected from aquatic products for entry and exit. The detection results were consistent with those of SN reference methods. It took less than 8h to detect these three pathogenic bacteria qualitatively or quantitatively in samples of aquatic products, and it could be applied to the quick detection of these three pathogenic bacteria in aquatic products for entry and exit.
Keywords：SalmonellasppVibrioparahaemolytichusEscherichiaColiOt57： H7MultiplexReal-TimePCR
Publication Date：2011-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 22-27 )