肠道致病菌PCR检测与鉴定研究

Detection and identification of pathogenic enterobacteria by PCR

目的 为了实现对腹泻等病原菌的快速、准确检测与鉴定 ,对引起感染性腹泻的暴发与流行的常见肠道致病菌进行检测与鉴定方法研究。方法 将常规PCR与半套式PCR及随机引物扩增DNA多态性分析 (RAPD)等技术相结合 ,对常见肠道致病菌 ,如志贺菌、沙门菌和致病性大肠杆菌O15 7∶H7等进行检测与鉴定。结果 uidA引物可特异扩增沙门菌、志贺菌及大肠杆菌 ,而 3种特异性引物则只扩增相应的致病菌 ;第一次PCR敏感性为 30～ 5 0CFU ,半套式PCR的敏感性可达 3～5CFU。结论 该技术特异性强、敏感性高、操作简便、快速 ,是环境样品及临床腹泻等病原学检测与诊断的有效方法之一。

Objective Pathogenic enterobacteria are the most frequent etiologic agents of infectious enteritis. In order to make a rapid and definite diagnosis of bacterial enteritis, a set of conventional PCR assay was developed to detect and identify Shigella, Salmonella and E. coli O157∶H7 directly from fecal or environmental samples. Methods The detection system included common PCR, semi PCR and RAPD. Results The uidA primer could be used to detect Shigella, Salmonella and E.coli O157∶H7, and the species primer could only be used to amplify the corresponding bacteria; the detecting limit for PCR was 30 to 50CFU, and the sensitive for semi PCR was 3 to 5 CFU. Conclusions Not only is this method more sensitive, specific and efficient, but also the processing is rapid and simple. This PCR method, therefore, will be a routine and practical protocols for detecting and identifying pathogenic microorganisms from clinical or environmental samples.