应用双重PCR检测产毒素多杀性巴氏杆菌  

Detection of Toxigenic Pasteurella multocida by Diplex PCR

作　　者：唐先春[1] 吴斌[1] 刘国平[1] 刘建杰[1] 王大鹏[1] 陈焕春[1] 

机构地区：[1]华中农业大学动物病原微生物实验室,湖北武汉430070

出　　处：《中国兽医学报》2005年第4期374-375,共2页Chinese Journal of Veterinary Science

基　　金：湖北省科技攻关项目(2001AA201)

摘　　要：参照有关文献设计了2对引物,分别扩增多杀性巴氏杆菌的Kmt及toxA基因,以使同时检测并区分产毒素与非产毒素多杀性巴氏杆菌。该PCR能检出103cfu菌量的模板。特异性试验表明,这2对引物都不能从猪的其他6种常见病原菌扩出特异性的带。对扩增的2个PCR产物测序结果表明,2序列都有很高的保守性,从而进一步证明了该PCR方法的特异性及灵敏性。临床应用,从386份病料分离出66株多杀性巴氏杆菌,其中有8株均能同时扩出457bp和864bp的片段,而其他54株只能扩出457bp的片段。Two sets of specific primers which amplify Kmt and ToxA gene of Pasteurella multocida separately were designed,so as to detect and identify toxigenic and nontoxigenic P.multocida at the same time.As little as 103 cfu of T+Pm template can be detected by this PCR.Specificity test showed that the two sets of primers can not amplify DNA products from 6 other swine pathogenic bacteria.The sequencing result of the two PCR products indicated that Kmt and toxA gene were quite conservative.It further confirmed the specificity and sensitivity of this diplex PCR.Sixty-six P.multocidas were isolated by using this PCR,within them,8 isolates can amplify both 457 base pair and 864 base pair DNA products,and 54 isolates can only amplify 457 base pair DNA product.

关 键 词：产毒多杀性巴氏杆菌 PCR 检测 

分 类 号：S852.61[农业科学—基础兽医学]