用PCR鉴定大肠杆菌O157∶H7  

Identification of Escherichia coli O157H7 by PCR

作　　者：周志江[1,2] 黄上媛[1,2] 郑明光[1,2] 汪力亚 王丽 李景云[1,2] 

机构地区：[1]解放军农牧大学兽医学院 [2]解放军302医院微生物研究室

出　　处：《中国兽医学报》1999年第3期248-250,共3页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金

摘　　要：根据大肠杆菌Ｏ１５７∶Ｈ７的编码ｅａｅ蛋白的ｅａｅＡ基因和大肠杆菌编码Ｈ７抗原的ｆｌｉＣ基因的核甘酸序列，合成了２对寡核苷酸引物，建立了一个检测大肠杆菌Ｏ１５７∶Ｈ７的ＰＣＲ方法。对１１株已知大肠杆菌Ｏ１５７∶Ｈ７（ＮＭ；无运动性）株和其他不同属的４２株已知肠道致病菌的检测结果表明，该方法只从大肠杆菌Ｏ１５７∶Ｈ７（ＮＭ）株的ＤＮＡ中产生预期的扩增产物，而从其他菌株的ＤＮＡ中未扩增出任何ＤＮＡ产物。该方法从基因水平直接确定大肠杆菌的血清型，特异性强，克服了以往血清学方法有非特异性反应的缺陷，为检测和鉴定大肠杆菌Ｏ１５７∶Ｈ７（ＮＭ）提供了一个新方法。Two sets of oligonucleotide primers derived from the sequences of the eae A gene of Escherichia coli O157H7 and the fliC gene of Escherichia coli respectively were synthesized, and a polymerase chain reaction (PCR) method was developed for the identification of Escherichia coli O157H7. A total of 10 Escherichia coli O157H7 (NM) and 43 other recognized enteropathogenic bacteria of several genera were studied. The expected PCR amplification products were observed only in DNAs from recognized Escherichia coli O157H7 (NM), and no amplification product was observed in the template DNAs extracted from other enteric pathogens. This method was able to determine the Escherichia coli O157H7 serotype directly on the basis of specific genes, and showed a high specificity. This PCR method may be a new way for the detection and identification of Escherichia coli O157H7.

关 键 词：聚合酶链反应 大肠杆菌 O157:H7 鉴定 PCR 

分 类 号：S852.612[农业科学—基础兽医学]