珠江河口水体霍乱弧菌监测及菌株毒力基因分布特征  

Surveillance program on and the distribution related to the virulence-associated genes of Vibrio cholerae in estuary of Pearl River

作　　者：李柏生[1] 王多春[2] 谭海玲[1] 柯碧霞[1] 陈经雕[1] 何冬梅[1] 刘美真[1] 邓小玲[1] 柯昌文[1] 阚飙 

机构地区：[1]广东省疾病预防控制中心病原微生物检验所,广州510300 [2]中国疾病预防控制中心传染病预防控制所

出　　处：《中华流行病学杂志》2011年第12期1242-1246,共5页Chinese Journal of Epidemiology

基　　金：国家高技术研究发展计划(863计划)，国家自然科学基金，国家科技重大专项，广州市医药卫生科技重大项目

摘　　要：目的了解珠江河口水体中01群和0139群霍乱弧菌的分布状况，分析菌株的分子特征和毒力基因特征。方法对2009年1月至2010年12月从珠江河口水体中分离的59株01群和10株0139群霍乱弧菌，采用聚合酶链反应（PCR）方法体外检测ctxA、fc硝、ace、zot、tcpl,hlyA、toxR和ompU等毒力相关基因，并进行毒力相关基因分型分析，对限制性内切酶NotI消化后的基因组DNA进行脉冲场凝胶电泳（PFGE）分析，采用BioNumerics软件分析图谱，得到菌株带型相似性的聚类分析树状图。结果2009--2010年共采集1152份水体标本，分离得到0I／0139群霍乱弧菌69株，其中O1群埃尔托霍乱弧菌59株（小川型18株，稻叶型41株），0139群霍乱弧菌10株。PCR检测69株菌ctxA全部阴性，hlyA和toxR全部阳性，基因分型可分成9个型。稻叶型菌株中，34．15％（14／41）为hlyA＋toxR＋ompU＋ace＋zot＋tcpI＋型；小川型菌株中，66．67％（12／18）为hlyA＋toxR＋型；0139群菌株中，70％（7／10）为hlyA＋toxR＋型。PFGE分型发现，0139群菌株PFGE相似度为69．9％～95．5％；01群菌株相似度为72．8％。100．0％，可分成3个聚类。结论在霍乱流行间歇期，该地区外环境水体中01群和0139群霍乱弧菌非产毒株广泛存在，基因型别多样。Objective To understand the distribution, molecular characteristics and virulence genes of the O1 and O139 Vibrio cholerae isolates from the Pearl River Estuary water. Methods Vibrio cholerae isolates collected from the Pearl River estuary waters from January 2009 to December 2010, were tested by PCR for eight virulence-related genes, including cholera toxin （ctxA） , zonula occludens toxin （zot）, accessory cholera enterotoxin （ace）, hemolysin （hlyA）, toxin-coregulated pilus （tcpA） , outer membrane protein （ompU） , and the regulatory protein genes （tcpl, toxR）. Genetic relation was assessed by pulsed-field gel electrophoresis （PFGE） and the patterns were clustered by BioNumerics. Results From 1152 aquatic samples, 69 isolates were identified, including 41 Inaba, 18 Ogawa and 10 O139. All the isolates showed ctxA negative, while the hlyA and toxR genes were positive in all the isolates. 34.15% （14/41） of the Inaba strains were hlyA＋ toxR＋ ompU＋ ace＋ zot＋ tepl＋, while 66.67% （12/18） belonged to Ogawa strains and 70% （7/10） of the O139 strains were hlyA＋ toxR＋. Through PFGE analysis, the O1 isolates formed three clusters in this study. The patterns of O1 isolates differed widely, with the similarity as 72.8%-100.0%, while the patterns of O139 isolates having the similarity of 69.9%-95.5%. Conclusion The non-toxigenic O1 and O139 V. cholerae had a wide distribution in the environment of Pearl River estuary water during the nonepidemic period of cholera. All the aquatic isolates presented diversities on the related virulent genes.

关 键 词：霍乱弧菌 毒力基因 脉冲场凝胶电泳 

分 类 号：R378[医药卫生—病原生物学]