Establishment of rapid detection method for Vibrio vulnificus in aquatic food by loop-mediated isothermal amplification.

Vibrio vulnificus is an important foodborne pathogen,which mainly exists in estuaries and the marine environment and seriously harms aquaculture development and human health.Therefore,the development of a technique that can identify the pathogen quickly,accurately and protect aquaculture development and enhance the food safety is of great importance.Here a rapid method for detecting Vibrio vulnificus with loop-mediated isothermal amplification(LAMP)using the vvHA gene of Vibrio vulnificus as target gene were established.Forty-six bacterial strains were selected to test specificity of the LAMP method and only Vibrio vulnificus strain was LAMP positive result,and indicating the high specificity of the LAMP method.The LAMP method could be completed in 40~60min and showed a good sensitivity of 15 CFU/ml for cultured Vibrio vulnificus and of 20 CFU/g for its contaminated food.Finally,25 samples in detected 278 practical samples of shrimps,crabs,oysters and artificial contamination ones were positive results using the LAMP method,which accorded with the detection results according to SN/T 1870-2007.The LAMP assay is a sensitive,rapid simple tool for the detection of Vibrio vulnificus.