Development of a dual real-time PCR assay for rapid simultaneous detection of Salmonella spp. and Vibrio parahaemolyticus

A dual real-time PCR assay was developed for the simultaneous detection of Salmonella spp. and Vibrio parahaemolyticus using primers and probe designed for the conservative domain of invA gene of Salmonella spp. and toxR gene of V.parahaemolyticus.The method was highly reproducible,specific and sensitive for detection of Salmonella spp. and V.parahaemolyticus,which could detect simulta-neously less than 10 CFU Salmonella spp. and V.parahaemolyticus in the same reaction system.In the specificity test,29 non-target reference and standard strains were negative.All the variation coefficients of intra-assay and inter-assay were less than 2% in the quantitative tests.The assay was applied to samples experimentally-contaminated with the two pathogenic bacteria and to aquatic products for entry and exit,and the results were consistent with those of SN standards.It took less than 8 h to qualitatively or quantitatively detect Salmonella spp. and V.parahaemolyticus in samples simultaneously.