霍乱弧菌多重PCR-DHPLC分型方法建立  

Development of multiplex PCR-DHPLC typing method for Vibrio cholerae

作　　者：黄晓蓉[1] 邵碧英[1] 王传得[2] 郑晶[1] 曹际娟[3] 陈彬[1] 吴谦[1] 

机构地区：[1]福建出入境检验检疫局技术中心微生物室,福州350001 [2]福建农林大学动物科学院 [3]辽宁出入境检验检疫局

出　　处：《中国公共卫生》2009年第6期721-722,共2页Chinese Journal of Public Health

基　　金：国家“十一五”科技支撑计划项目子课题(2006BAK02A13-11)

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。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 multi- plex 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.

关 键 词：霍乱弧菌 多重PCR 变性高效液相色谱(DHPLC) 

分 类 号：Q93-332[生物学—微生物学]