犊牛腹泻主要病原菌多重PCR方法的建立

Multiplex PCR for Major Pathogenic Bacteria Causing Diarrhea in Calves

产毒性大肠埃希菌、A/E大肠埃希菌和沙门菌是造成犊牛腹泻的主要病原菌。选取产毒性大肠埃希菌的st和lt基因、A/E大肠埃希菌的eae基因和沙门菌的invA基因作为扩增靶基因序列,通过优化反应条件建立了四重PCR检测体系。对PCR产物回收、测序,验证PCR。通过特异性试验和敏感性试验证明该PCR体系特异性强、敏感性高,可有效地检测犊牛腹泻病原菌。使用该四重PCR检测22份临床样品,发现其中9份携带相关基因,并分离得到了相关致病菌。

Enterotoxigenic Escherichia coli,attaching and effacing E.coli,and Salmonella are major pathogenic bacteria causing diarrhea in calves.A multiplex PCR was developed to identify the bacteria by amplifying genes of eae carried by AEEC,invA carried by Salmonella,st and lt carried by ETEC.The multiplex PCR was verified by sequencing the products of PCR.The multiplex PCR was specific and sensitive and is efficient to detect the pathogenic bacteria.Nine of 22 clinical samples carrying related genes were detected by this multiplex PCR.