应用多元PCR鉴定水体中大肠杆菌的毒素基因

Detection for Virulence Genes of Escherchia coli Isolates Recovered from Water Body Using Multiplex PCR

根据已发表的大肠杆菌(Escherchia coli)毒素基因序列,针对大肠杆菌的8种毒素基因设计特异引物并进行多元PCR扩增,检测某猪场排污口附近分离的16株大肠杆菌的毒素基因。结果表明,从16株待测样品中成功扩增出7个毒素基因,即fanA、fedA、aidA-1、stx2e、astA、fasA和sepA。其PCR产物大小与预期一致。在16个样品中,aidA-1的检出率最高,总共16株。其后依次为stx2e 9株,astA 8株,fanA 7株,sepA 2株,fedA和fasA各1株。实验成功验证了多元PCR技术的可靠性及其在水环境中大肠杆菌毒素基因检测的实用性。

According to the published DNA sequences,eight pairs of specific oligonucleotide primers were designed to identify 8 virulence genes harbored by 16 Escherchia coli isolates recovered from water body of a pig farm using multiplex PCR method.7 virulence genes（fanA,fedA,aidA-1,stx2e,astA,fasA and sepA） were detected in the strains.All the strains harbored aidA-1 gene while stx2e,astA,fanA,sepA 2,fedA and fasA were detected in 9,8,7,2,1 and 1 strains.The study successfully demonstrated the reliability of multiplex PCR technique and its practicability in detection of virulence genes of E.coli within aquatic environment.