The Molecular Detection Method For Vibrio Cholerae, Vibrio Parahaemolyticus And Vibrio Vulnificus In Seafood Samples By Using Multiplex PCR In Macao

The microbiological safety of food is always an important concern of global health. In fact, there are many food-borne illness caused by bacterial pathogens every year in Macao. Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus are the common bacteria, which always involve in the cases of seafood poisoning. Therefore, the effective prevention of the bacterial food-borne disease, a rapid and reliable detection method of the bacterial pathogens in food is acting a very important role. The conventional detection method of Vibrio cholerae 01/0139 and Vibrio parahaemolyticus in seafood samples are usually used in food laboratories of Macao. This method is time-consuming and laborious, so there is a need for the development of innovative methods for the rapid detection and identification of Vibrio spp. in seafood samples.There were 76 seafood samples have been examined by using conventional culture method and multiplex PCR (mPCR) method from May to October 2006. The main sample source was purchased from the open markets, supermarkets, restaurants within Macao and the other source was come from the beaches and coast sides in Macao. The conventional culture method and the mPCR assay were modified from the methods mentioned in Bacteriological Analytical Manual Online published by Food and Drug Administration (1). The test of mPCR detection limit has been performed in this study, and the result shows that 10~1 cells of V. parahaemolyticus, 10~2 cells of V. vulnificus and 10~3 cells of V. cholerae 01/0139 can be detected by the mPCR assay.In the results of sample detections, even though there was no toxigenic V. cholerae detected in these experiments but the high positive rate of V. parahaemoltyicus (98.7%) and V. vulnificus (71.1%) were found by mPCR method in the samples. Whereas, the isolation rate of this three Vibrio spp. from the culture method was low (toxigenic V. cholerae 0%; V. parahaemolyticus 46.1% and V. vulnificus 21.1%). The low isolation rate of culture method could be due to...