Development of multiplex PCR-DHPLC typing method for Vibrio cholerae

Objective To develop a tpyingmethod ofmultiplex PCR-DHPLC forVibrio cholerae.Metbods The special priners were composed to expand the collagenase gene(vcc gene) of Vibrio cholerae,the virulent genes of V.cholerae O 1 and O 139(ctxA gene and tcpA gene),and the virulent gene of V.cholerae O 139 (LPSgt gene),respectively.The multiplex PCR-DHPLC typing method was developed after the PCR anneal temperature for four pairs of grin ers being optinized.Results The fourgeneswere amplified synchronously. The typing results of V.cholerae bymultiplex PCR-DHPLC method were the same as those of anticipated.Conclusion It is rapid and exact to identify whether the bacteria pure culture is V.cholerae and idiographic strain according to themultiplex PCR-DHPLC detection result.