摘要
目的 建立简易实用的检测EHEC和EPEC多种毒力基因的方法。方法 应用多重PCR同时检测大肠埃希菌菌株的stx1、stx2、EPEC和EHEC共同的eaeA(eaeA -gen)等靶基因 ;用常规PCR检测EHECO15 7特异的eaeA(eaeA -O15 7)基因 ,以区分EHEC和EPEC的eaeA基因。结果 对分别分离自美国和我国江苏的EHECO15 7菌株以MPCR检测 ,stx1、stx2和eaeA -gen基因均阳性 ,同时常规PCR扩增eaeA -O15 7也阳性。在 8株分离自杭州的志贺毒素阴性的大肠埃希菌O15 7菌株中 ,stx1、stx2和eaeA -O15 7基因均阴性 ,仅 2株扩增出EPECeaeA基因。在 2 8株EPEC血清型、12株ETEC血清型、12株EIEC血清型大肠埃希菌菌株中 ,stx1和stx2基因均阴性 ;3株EPEC血清型菌株eaeA - gen基因阳性。结论 MPCR技术可同时检测菌株的stx1、stx2和eaeA基因 ,可为EHEC和EPEC感染的诊断及流行病学调查 ,提供一种简便、实用。
Aim To develop a simple, efficient method for determining the virulence trait of enterohemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC). Methods Virulence genes, including stx1, stx2, EPEC and EHEC general eaeA fragment (eaeA-gen), were amplified by multiplex PCR in E. coli strains.In order to distinguish EHEC eaeA from EPEC eaeA, another PCR was used to amplify EHEC O157 specific eaeA fragment (eaeA-O157). Results In EHEC O157∶H7 strain 933 and 88-2364,isolated from US and Jiangsu in China respectively, stx1, stx2, eaeA-gen and eaeA-O157 genes were all found by the multiplex PCR. In 8 strains of nontoxinogenic E. coli O157 isolated from Hangzhou, there was no stx1, stx2, or eaeA-O157 genes, except 2 strains isolated from diarrhea patients showed EPEC eaeA gene positive. Of all 28 E. coli strains with EPEC O serotype which were showed both of the stx1 and stx2 negative, 3 were EPEC eaeA gene positive. Neither EHEC nor EPEC virulence gene was found in all tested E. coli strains with ETEC and EIEC O serotypes.Conclusion The multiplex PCR can be used to detect stx1, stx2, and eaeA genes at the same time.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2001年第1期52-55,共4页
Chinese Journal of Zoonoses
