直接酶联免疫吸附法和聚合酶链反应联合检测沙门菌的应用

目的　通过比较试验 ,得到准确、快速的沙门菌检测方法。方法　 2 2 8株沙门菌经过前增菌、选择性增菌后 ,采用在聚合酶链反应 (PCR)和酶联免疫吸附法 (ELISA)同时检测 ,并将 2种快速检测方法进行比较研究。结果　PCR法的敏感性优于直接ELISA法 ,2种方法的检测符合率达 99%以上。直接ELISA结合PCR法对饮食行业工作人员健康检查的 15 4 6份人粪便样品进行检测 ,同时以国家标准方法为参照 ,直接ELISA法的敏感性和特异性达 10 0 %和 97 14 % ,PCR法的敏感性和特异性均为 10 0 %。结论　优化的沙门菌检测程序是对大量样品采用直接ELISA筛检 ,除去大量阴性样品 ,阳性样品用PCR法作进一步鉴定 ;血清型的确定用国家标准方法。

Combination of direct-ELISA and PCR for the rapid detection and identification of Salmonella spp

Objective To develop a protocol for the rapid detection of Salmonellae Methods A mono-antibody-based direct-ELISA and PCR methods for the detection of Salmonella were developed previously. This study assessed the accuracy of both direct-ELISA and PCR methods for the rapid detection of Salmonella and set up a new detection protocol. Results The sensitivity of the PCR method was higher than that of direct-ELISA method. In the 2002 spring physical examination for employees, 1 546 human fecal samples were examined by the combination of direct-ELISA and PCR method. Compared with the results of national standard method, the sensitivity and specificity of direct ELISA was 100% and 97.14%, respectively, while those of PCR method reached both 100%. It also indicated that combination use of two methods could give positive report within 40 hrs, and also achieve high sensitivity and specificity. Conclusions Based on the results obtained, a protocol for the rapid detection of Salmonella was developed. The first step is to us direct-ELISA method to screen the large number of samples, and then use PCR method to validate the ELISA positive samples, and the final step is, if needed, is to use the national standard method to determine the serotypes of Salmonellae.