四川地区鸡源产气荚膜梭菌的分离鉴定与基因分型  

Isolation,identification and genotyping of Clostridium perfringens from chickens in Sichuan province

作　　者：倪学勤[1,2] 宋振银[1,3] 曾东[1,2] 郑晓丽[1,3] Joshua Gong 

机构地区：[1]四川农业大学动物医学院动物微生态研究中心,雅安625014 [2]动物疫病与人类健康四川省重点实验室,雅安625014 [3]四川农业大学资源环境学院,雅安625014 [4]Food Research Program,Agriculture and Agri-Food Canada

出　　处：《中国人兽共患病学报》2009年第8期737-740,786,共5页Chinese Journal of Zoonoses

基　　金：教育部创新团队项目(IRT0848)资助;四川省科技厅国际合作项目(HH20080016)资助

摘　　要：目的检测四川地区健康鸡群产气荚膜梭菌的分布状况。方法从四川地区规模化鸡场采集新鲜粪便样品,按产气荚膜梭菌α、β、ε、ι毒素基因cpa、cpb、etx及iA序列,设计针对4种毒素基因的4对特异引物,应用多重PCR方法对产气荚膜梭菌进行基因分型。最后利用重复序列PCR进一步做亚型分析。结果从150份样品中分离到8株(5.3%)产气荚膜梭菌,多重PCR都只扩增出α条带,毒素基因分型结果均为A型。ERIC-PCR、REP-PCR图谱显示两者均适用于产气荚膜梭菌亚型分析,ERIC-PCR是一种更简便、快速和有效的分子流行病学调查方法。结论四川地区健康鸡群中存在的产气荚膜梭菌主要是A型。To analyze the topography and gene type of C. perfringens in healthy chickens of Sichuan province. Fresh fecal samples from integrated poultry farms of Siehuan province were collected. C. perfringens were isolated with euhure method and identified with biochemical test. Then multiple PCR was developed with 4 sets of primers designed based on the sequences of 4 C. perfringens toxin genes （cpa, apb, etx and iA） to genotype the identified isolates. Finally, repetitive-element PCR was used to analyze subtype of C. perfringens. Eight isolates were obtained from 150 samples and were genotyped as type A by multiple PCR. ERIC PCR and REP PCR were found to be equally suitable for subtyping of C. perfringens isolates. ERIC-PCR proved to be an easy-to-perform and relatively fast method which can serve as an effective mean for molecular epidemiology. C. perfringens type A is the main Clostridium species in the healthy chickens in Sichuan province.

关 键 词：产气荚膜梭菌 多重PCR 基因分型 鸡 

分 类 号：R378.3[医药卫生—病原生物学]