检测987p+肠毒素性大肠埃希氏菌PCR方法的建立

以 98 7p菌毛结构基因保守序列为靶序列 ,设计合成了 1对可扩增 45 9bp目的片段的引物 ,建立了检测肠毒素性大肠埃希氏菌 987p菌毛基因的PCR方法。该方法对K 88 ( 为菌毛阳性 )、K 99 、F 41 参考菌株和链球菌、葡萄球菌、巴氏杆菌的检测结果均为阴性 ;该方法的敏感度可达 10 2 CFU ,对 2 0株腹泻仔猪粪便分离物进行检测 ,有 1株为阳性 ,与血清学检测的结果一致。结果表明 ,此方法特异性和敏感性都很高 ,可用于临床 987p肠毒素性大肠埃希氏菌病的快速诊断和流行病学调查

Detection of 987p positive Enterotoxigenic Escherichia coli by Polymerase Chain Reaction

p fimbriae is one of the adhesive fimbriae antigens of enterotoxigenic Escherichia coli. A polymerase chain reaction(PCR) method was developed to detect 987p positive Escherichia coli by means of amplifing 987p fimbriae gene. The primer used in this assay was designed from the conserved section of 987p structural gene sequences. The detection specificity of this PCR method was confirmed with PCR assay of enterotoxigenic Escherichia coli reference strains, Salmonella typhimurium, Streptococcus, Staphylococcus aureus as well as Pasteurella pneumotropica. 20 Escherichia coli strains isolated from 1 to 7 days old diarrhea piglets were detected with this PCR method, and one of these is positive for the enterotoxigenic property. And the same result was achieved with slide agglutination.The results demonstrated that this PCR was specific and sensitive. It will be useful for identification of 987p positive Escherichia coli strains.