Abstract：A multiplex PCR method was developed for rapid and specific detection of Escherichia coli O157︰H7 and three of its virulence genes （hylA, eaeA and stx2 gene）.Five sets of primers were designed according to the sequences of rfbE gene、fliC gene、hemolysin gene（hlyA）、inthnin gene (eaeA) and shiga-like toxin gene 2（stx2）of E.coli O157 were selected and added into one amplification system to perform PCR，The system was optimized, the specificity and sensitivity of this system were evaluated， and used to detect the clinical isolates. Results: The assay was designed to amplify the 327 bp、247 bp、494 bp、384 bp、and 779 bp regions of corresponding genes rfbE、fliC、hlyA、eaeA and stx2. It was sensitive and specific highly. Sensitivity of the assay was 104cfu /ml of bacteria samples. Conclusions: A rapid, specific, and sensitive multiplex PCR technique for the simultaneous detection of E.coli O157︰H7 and three of its virulence genes has been studied primarily. And available to test for the quantitative detection of E.coli O157︰H7 from the contaminated food samples.
2013, Vol. 29 
