直接ELISA和PCR相结合快速检测样品中的沙门氏菌

目的 通过样品试验，得到优化的沙门氏菌检测方法。方法 在单抗直接ELISA和PCR法检测沙门氏菌的基础上，将PCR法和单抗直接ELISA两种快速检测方法进行比较研究。结果 直接ELISA方法结合PCR方法对城区141份水样、2002年春季饮食行业工作人员健康检查的1426份人粪便样品，并与常规国标方法对比，直接ELISA法的敏感性和特异性达100％和97．2％，PCR法的敏感性和特异性均为100％。通过对鲜牛奶样、龙虾、虾仁、熟食制品、水样、人粪样测试，最终确定较优化的沙门氏菌检测程序。结论 PCR法的敏感性优于直接ELISA法。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；需要确定血清型时则用国标方法。

Combination of direct ELISA and PCR for rapid detection of Salmonella

Aim An opitimized rapid protocol for the detection of Salmonella was developed.Methods An Mabbased direct ELISA and PCR protocol for the detection of Salmonella were developed previously.This study investigated the accuracy of both direct ELISA and PCR for the rapid detection of Salmonella and set up a new detection strategy.Result Using directELISA to detect water samples from downtown area of Yangzhou city,and then using PCR to test the ELISA positive samples and randomly selected negative samples,the coincidence of their results was 97.2%.In the 2002 spring physical examination of Yangzhou Guangling district for employees in the restaurant industry,1426 human feces samples were detected by the combination strategy of directELISA and PCR method.Compared with the results of National Standard method,the sensitivity and specificity of direct ELISA was 100% and 97%,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.Based on all the results,an opitimized rapid protocol for the detection of Salmonella was developed.Conclusion The results suggest that the sensitivity of PCR is higher than that of directELISA.use directELISA to screen the large number of samples first,and then use PCR to test the ELISA positive samples,the final step is,if needed,to use National Standard method to determine the serotypes of Salmonellae.