Establishment and application of multiple PCR technique for the rapid detection of Salmonella sp.,Shigella sp. and Vibrio parahaemolyticus

Objective To establish a multiplex PCR technique for the simultaneous detection of Salmonella sp.(SM),Shigella sp.(SP) and Vibrio parahaemolyticus.(VP).Methods Three sets of primers were designed to amplify the gene segments of hilA of Salmonella sp.,ipaH of Shigella sp.,and tdh of Vibrio parahaemolyticus,and the products were analyzed by electrophoresis.At the same time,this system was optimized,and the specificity and sensitivity of this system were evaluated.Results Three target bacteria were detecte d in 8 h by using this multiple PCR technique.The high specificity was demonstrated by detecting other nine kinds of food-borne pathogenic bacteria.and the detection sensitivity of multiplex PCR were 104 cfu/mL for SM,102 cfu/mL for SP,104cfu/mL for VP.Artificially contaminated food and stool samples were inspected with this method.Conclusions A rapid,specific,and sensitive multiplex PCR technique for the simultaneous detection of Salmonella sp.,Shigella sp.and Vibrio parahaemolyticus.in 8 h has been studied primarily.