Meng J, Zhao S, Doyle M P, Mitchell S E, Kresovich S
Center for Food Safety and Quality Enhancement, University of Georgia, Griffin 30223-1797, USA.
Lett Appl Microbiol. 1997 Mar;24(3):172-6. doi: 10.1046/j.1472-765x.1997.00375.x.
A multiplex PCR assay specifically detecting Escherichia coli O157:H7 was developed by employing primers amplifying a DNA sequence upstream of E. coli O157:H7 eaeA gene and genes encoding Shiga-like toxins (SLT) I and II. Analysis of 151 bacterial strains revealed that all E. coli O157:H7 strains were identified simultaneously with the SLT types and could be distinguished from E. coli O55:H7 and E. coli O55:NM, and other non-O157 SLT-producing E. coli strains. Primer design, reaction composition (in particular, primer quantity and ratios), and amplification profile were most important in development of this multiplex PCR. This assay can serve not only as a confirmation test but also potentially can be applied to detect the pathogen in food.
通过使用引物扩增大肠杆菌O157:H7 eaeA基因上游的DNA序列以及编码志贺样毒素（SLT）I和II的基因，开发了一种特异性检测大肠杆菌O157:H7的多重PCR检测方法。对151株细菌菌株的分析表明，所有大肠杆菌O157:H7菌株都能与SLT类型同时鉴定出来，并且可以与大肠杆菌O55:H7、大肠杆菌O55:NM以及其他非O157产SLT的大肠杆菌菌株区分开来。引物设计、反应组成（特别是引物数量和比例）以及扩增曲线在该多重PCR的开发中最为重要。该检测方法不仅可以作为确认试验，还可能用于检测食品中的病原体。
