Rapid detection of Vibrio parahaemolyticus in seafood by Real- time PCR targeted to the H- NS gene

Objective: In this study,a rapid and accurate method was established to detect Vibrio parahaemolyticus of using SYBR Green I fluorescence PCR reaction.Methods: Primers were designed by using conserved region of the H- NS gene.Twelve non Vibrio parahaemolyticus were amplified by PCR to confirm the specificity.Based on H- NS gene,the standard curve was established and the sensitivity was determined by SYBR Green I fluorescence PCR reaction. Sterile clam samples contaminated by different concentrations of bacteria were detected to determine the feasibility of this method.Then,thirty clam samples were detected,and the results were compared with the GB 4789.7-2013.Results: Correlation coefficient of the standard curve established in this study was 0.998,and the sensitivity was 7.3 × 10 CFU / m L.The detection limit of artificial contamination was 6.7 × 102 CFU / m L.There were four positive results detected in the thirty samples by using SYBR Green I fluorescence PCR method which was consistent with GB 4789.7- 2013.Conclusion: Method of this article could be used in rapid detection of Vibrio parahaemolyticus.