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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 In order to achieve rapid and accurate detection and identification of pathogens such as diarrhea, the detection and identification of common enteric pathogens causing outbreak and epidemic of infectious diarrhea were studied. Methods By combining conventional PCR with semi-nested PCR and random amplified polymorphic DNA (RAPD) and other techniques, the common pathogenic bacteria such as Shigella, Salmonella and pathogenic Escherichia coli O157 : H7 and other testing and identification. Results The uidA primers could specifically amplify Salmonella, Shigella and Escherichia coli, while the three specific primers only amplified the corresponding pathogenic bacteria. The sensitivity of the first PCR was 30 ~ 50CFU, Sensitivity up to 3 ~ 5CFU. Conclusion The technique is characterized by high specificity, high sensitivity, simple and rapid operation, and is an effective method for the detection and diagnosis of environmental samples and clinical diarrhea.