【摘要】 目的建立霍乱弧菌多重PCR-变性高效液相色谱(DHPLC)快速分型方法。方法分别合成扩增霍乱弧菌胶原酶基因(vcc基因)、O1群和O139群的毒力基因(ctxA基因和tcpA基因)以及O139群毒力基因(LPSgt基因),并对4对引物的PCR退火温度进行优化,建立多重PCR-DHPLC分型方法。结果4种基因可同时被扩增,应用多重PCR-DHPLC方法对霍乱弧菌进行分型的结果与预期的一致。结论多重PCR-DHPLC分型方法可用于快速、准确地鉴定细菌纯培养物是否为霍乱弧菌以及具体的菌群。更多还原

【Abstract】 Objective To develop a tpying method of multiplex PCR-DHPLC for Vibrio cholerae.Methods The special primers were composed to expand the collagenase gene(vcc gene) of Vibrio cholerae,the virulent genes of V.cholerae O1 and O139(ctxA gene and tcpA gene),and the virulent gene of V.cholerae O139 (LPSgt gene),respectively.The multiplex PCR-DHPLC typing method was developed after the PCR anneal temperature for four pairs of primers being optimized.Results The four genes were amplified synchronously.The typing results of V.cholerae by multiplex 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 the multiplex PCR-DHPLC detection result.更多还原