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犬产气荚膜梭菌的分离和PCR检测方法的建立

彭建国， 强京宁， 方建强

（公安部昆明警犬基地， 云南 昆明650204）

摘要： 从12头疑似产气荚膜梭菌感染的猝死警犬中分离获得了18株病原菌，经生化试验及毒素中和试验，确定为A型和C型产气荚膜梭菌。参考GenBank中登录的基因序列，设计合成了针对产气荚膜梭菌α、β、ε、ι、CPE和β2 共6对分型毒素基因的引物，对以前昆明地区分离的1株产气荚膜梭菌进行了PCR扩增，结果扩增出了与预期大小相同的6个基因片段，分别为233、196、324、446、567和665bp。建立的PCR方法能同时用于产气荚膜梭菌鉴定和毒素分型，较细菌毒素检测方法快速，与细菌分离鉴定的结果一致。

关键词： 犬； 产气荚膜梭菌； 分离鉴定； PCR

中图分类号：S 852.616.3 文献标识码： A  文章编号：1000-6419(2004)12-0045-04

Isolation and development of PCR method for detection of Clostridium perfringens from dog

PENG Jianguo, QIANG Jingning, FANG Jianqiang

(Police Dog Base in Kunming， Ministry of Public Security, Kunming 650204，China)

Abstract: The 18 strains of Clostridium perfringens separated from 12 suddenlydied dogs infected with C.perfringens were identified as type A and C by the biochemical tests and the toxin neutralization reaction. Six couples of the specialized primers corresponding respectively to α,β,ε,ι, CPE and β2 of toxin gene were designed and synthesized according to the genetic sequences and the published literatures. The purpose fragments amplified by the PCR are 233, 196, 324, 446, 567 and 665bp in length respectively. The developed PCR method can be used in bacterial identification and toxin differentiation, and was more rapid than the method for detection of bacteriotoxin.
Key words: dog; Clostridium perfringens; isolation and identification; PCR

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