反刍动物源产气荚膜梭菌新疆流行株的分离鉴定及分型研究  

ISOLATION AND MOLECULAR TYPING OF CLOSTRIDIUM PERFRINGENS FROM RUMINANTS IN XINJIANG

作　　者：王晓婷 乔军[1] 孟庆玲[1] 蔡扩军[2] 李静[1] 李妍 李杰[1] 张国武 才学鹏[3] WANG Xiao-ting;QIAO Jun;MENG Qing-ling;CAI Kuo-jun;LI Jing;LI Yan;LI Jie;ZHANG Guo-wu;CAI Xue-peng(College of Animal Science and Technology,Shihezi University,Shihezi 832003,China;Animal Disease Control and Diagnosis Center in Urumqi,Urumqi 830063,China;China Institute of Veterinary Drugs Control,Beijing 100081,China)

机构地区：[1]石河子大学动物科技学院,石河子832003 [2]乌鲁木齐市动物疾病控制与诊断中心,乌鲁木齐830063 [3]中国兽医药品监察所,北京100081

出　　处：《中国动物传染病学报》2019年第6期22-28,共7页Chinese Journal of Animal Infectious Diseases

基　　金：兵团中青年科技创新领军人才计划(2016BC001);国课家重点研发计划子课题(2016YFD0500908-3)

摘　　要：为了弄清反刍动物源产气荚膜梭菌新疆流行株的生物学特性和分子特征,本研究从新疆不同地区规模化养殖场和屠宰场采集158份牛羊临床病料,进行产气荚膜梭菌的分离培养;通过形态学、生化特性、16S rRNA序列分析对产气荚膜梭菌分离株进行鉴定;利用多重PCR技术扩增产气荚膜梭菌α、β、ε、ι、β2基因,将测序结果与NCBI数据库比对进行新疆流行株的基因分型。结果从158份临床病料中共分离出产气荚膜梭菌67株,形态学、生化和16S rRNA序列分析鉴定结果均为产气荚膜梭菌。毒素基因检测表明,其中α基因检出率为100.00%(67/67),ε基因检出率为17.91%(12/67),β2基因检出率为37.31%(25/67),β、ι基因未检出。毒素基因分型可将产气荚膜梭菌新疆流行株分为A、D型,其中A型占82.09%(55/67),D型占17.91%(12/67),提示产气荚膜梭菌新疆流行株优势型别为A型。本研究为产气荚膜梭菌流行病学研究和防控提供科学依据。Total 158 clinical samples were collected from cattle and sheep on large-scale farms and slaughterhouses in Xinjiang to conduct the epidemiological investigation to Clostridium perfringens.The bacterial isolation and identification were performed based on morphology,biochemical characteristics and 16 S rRNA gene sequencing.The genes alpha,beta,epsilon,Iota,and beta2 of C.perfringens isolates were amplified by multiplex PCR.The sequencing data were compared with NCBI database for genotyping of C.perfringens epidemic isolates in Xinjiang.As a result,67 C.perfringens strains were isolated from these clinical samples and confirmed according to their morphology,biochemical and 16 S rRNA.Analysis of toxin genes of these strains showed thatαgene was positive in 67 stains(100.00%),εgene in 12 strains(17.91%)andβ2 gene in 25 strains(37.31%).βandιgenes were not detected in these strains.Toxin genotyping results showed that these C.perfringens strains belonged to genotypes A and D,among which 55 strains were genotype A and 12 strains were genotype D.This epidemiological investigation provided scientific information for prevalence and control of C.perfringens in Xinjiang.

关 键 词：产气荚膜梭菌 分离鉴定 反刍动物 

分 类 号：S852.616.3[农业科学—基础兽医学]