羊魏氏梭菌PCR方法的建立及初步应用  

Establishment and Application of PCR Assay for Clostridium perfringens Isolated from Sheep

作　　者：余波[1] 冉懋韬[1] 谭诗文[1] 徐景峨[1] 魏赐开 艾玉萍[1] 

机构地区：[1]贵州省畜牧兽医研究所,贵州贵阳550005 [2]贵州省瓮安县动物预控中心,贵州瓮安550400

出　　处：《中国畜牧兽医》2011年第11期106-108,共3页China Animal Husbandry & Veterinary Medicine

基　　金：贵州省农业科技攻关项目[黔科合NY字(2009)]3089号];贵州省畜禽健康养殖技术创新能力建设项目[黔科合院所创能(2010)4004号];中央补助地方科技基础条件专项基金项目[黔科条中补地(2010)4001];贵州省农科院专项[黔农科院院专项(2010)047号]

摘　　要：根据羊魏氏梭菌A、B、C、D、E型共有的α毒素基因,设计了1对引物,通过对PCR反应条件进行优化,建立了羊魏氏梭菌PCR检测方法。该方法扩增条带大小为255bp,最低核酸检测量A型为0.39ng/L、B型为0.62ng/L、C型为0.52ng/L、D型为0.87ng/L、E型为0.92ng/L,而对羊大肠杆菌、羊链球菌、金黄色葡萄球菌、羊多杀性巴氏杆菌的扩增结果均为阴性。应用该PCR方法对54份临床样本进行检测,PCR检测结果高于细菌学和生化检测结果。结果表明,该PCR方法具有很好的特异性和敏感性,可用于临床羊魏氏梭菌病的早期快速诊断。According to the gene sequences in GenBank of alpha-toxin,one pair of specific primer was designed for amplifying the specific fragments of alpha-toxin gene.After optimization of annealing temperature and primers concentrations,PCR assay was established for simultaneous detection of the Clostridium perfringens.The specific bands of 255 bp was amplified.The sensitivity and specificity tests showed that the PCR were sensitive in 0.39,0.62,0.52,0.87,0.92 ng/L A,B,C,D,EC.perfringens.No band was amplified from Escherichia coli,Streptococcus,Staphyloccocus aureus and Pasteurella multocida.54 clinical samples were detected by PCR.The results revealed that the established PCR assay was sensitive,specific and reproducible and it could be used to detect rapidly Clostridium perfringens isolated from sheep.

关 键 词：羊魏氏梭菌 Α毒素基因 快速诊断 

分 类 号：S858.26[农业科学—临床兽医学]