表型和分子分型技术在霍乱弧菌研究中的应用  

Application of phenotypic and molecular subtypic techniques in study of vibrio cholerae

作　　者：黄冰[1] 李孝权[1] 邓志爱[1] 张欣强[1] 

机构地区：[1]广州市疾病预防控制中心,510080

出　　处：《中国公共卫生管理》2012年第3期378-380,共3页Chinese Journal of Public Health Management

基　　金：广东省医学科研基金(A2010493);广州市医药卫生科技项目(2009-YB-121)

摘　　要：目的分析1999～2005年广州地区O1群和O139群霍乱弧菌代表菌株的致病基因型和PFGE型别,确定霍乱菌株的分子流行病学特征和同源性,研究本地霍乱弧菌的分子特性。方法采用多重PCR方法检测霍乱弧菌的4种致病相关基因,采用限制性内切酶Not I,以脉冲场凝胶电泳技术(PFGE)对菌株进行分子分型,采用分型处理软件BioNumerics Version对PFGE图谱做聚类分析。结果本地霍乱弧菌代表株中存在3种致病基因型(A、B、C型),病例分离的霍乱弧菌多为A型基因,而环境分离的霍乱弧菌则多为C型和B型。总共96株霍乱弧菌分为60个不同的PFGE型,带型范围:8 kb-650 kb。相同或相近的PFGE型(相差1～3条带)存在于多次霍乱暴发疫情分离株中、环境分离霍乱与临床株之间、输入病例与本地病例分离霍乱弧菌间,不同的PFGE克隆型(4～6带及以上)的流行病学联系也较远。结论将致病基因分型、PFGE型与流行病学资料结合,揭示了本地霍乱菌株从受污染的珠江水及水产品传递给人并引起霍乱暴发、散发的特征,提示开展霍乱菌株PFGE分子分型监测及加强地区间合作的必要性和急迫性。Objectives To analyze the virulence types and PFGE types of representative strains of Vibrio cholerae （V.cholerae） O1 and O139 serogroups isolated in Guangzhou area from 1999 to 2005 and to determine the molecularly epidemiological characteristics of these strains, in order to track the sources and changes of V.eholerae and study epidemic trerals and laws. Methods To apply Multiplex Polymerase Chain Reae- tion（MPCR） assay in detecting the four pathogenicity-related genes of V.eholerae. Pulse-field gel electrophoresis analysis（PFGE） using restric- tion enzyme Not I was employed in the molecularly subtyping of V.cholerae and PFGE patterns were analyzed by BioNumeries Version4.0 soft- ware. Results There are three virulence types in representative strains of V.cholerae O1 and O139 serogroups isolated in Guangzhou area, namely, virulence type A,B and C. Sixty distinct PFGE types in the range of 8-650kb were obtained among the 96 isolates typeable by PFGE. The same and closely related PFGE profiles with differences of 1-3 hands exist in the isolates in several cholera outbreaks, between isolates from the environment and human resources and also between isolates from imported cases and local cases as well. Heterogeneous PFGE clonal types with differences of as many as four bands and beyond indicated the distant relationship epidemiolgically. Conclusion The combina- tion of virulence types,PFGE elonal types and epidemiological data shown the epidemic trend that V.cholerae was transmitted from the contami- nated Pearl River water and seafood to human bodies triggering cholera outbreaks. It is necessary to reinforce cooperations between different re- gions in the PFGE typing of V.eholerae.

关 键 词：霍乱弧菌 多重PCR 致病相关基因 脉冲场凝胶电泳 分子分型 

分 类 号：R117[医药卫生—公共卫生与预防医学]