Fratamico P M, Sackitey S K, Wiedmann M, Deng M Y
Eastern Regional Research Center, U.S. Department of Agriculture, Philadelphia, Pennsylvania 19118, USA.
J Clin Microbiol. 1995 Aug;33(8):2188-91. doi: 10.1128/jcm.33.8.2188-2191.1995.
In order to develop a PCR assay for Escherichia coli O157:H7, a portion of the 60-MDa plasmid harbored by enterohemorrhagic E. coli (EHEC) was sequenced and PCR primers were designed. A multiplex PCR method was then designed by employing primers specific for the EHEC eaeA gene, conserved sequences of Shiga-like toxins I (SLT-I) and II (SLT-II), and the 60-MDa plasmid. PCR products of 1,087 bp (eaeA), 227 and/or 224 bp (SLT-I and/or SLT-II), and 166 bp (plasmid) were successfully amplified simultaneously in a single reaction. The multiplex PCR method can be used to specifically identify EHEC of serogroup O157.
为开发一种针对大肠杆菌O157:H7的聚合酶链反应（PCR）检测方法，对肠出血性大肠杆菌（EHEC）携带的60-MDa质粒的一部分进行了测序，并设计了PCR引物。然后通过使用针对EHEC eaeA基因、志贺样毒素I（SLT-I）和II（SLT-II）的保守序列以及60-MDa质粒的特异性引物，设计了一种多重PCR方法。在单个反应中成功同时扩增出了1087 bp（eaeA）、227和/或224 bp（SLT-I和/或SLT-II）以及166 bp（质粒）的PCR产物。该多重PCR方法可用于特异性鉴定O157血清型的EHEC。
