PCR快速检测沙门菌试剂盒的研制与应用

黄金林; 焦新安; 刘佩红; 张苏华; 潘志明; 文其乙; 刘秀梵

PCR快速检测沙门菌试剂盒的研制与应用

Development and application of PCR kit for rapid detection of salmonella

摘要

摘要:
目的在优化聚合酶链反应(PCR)检测沙门菌程序的基础上, 研制沙门菌PCR检测试剂盒。
方法通过样品检测, 确定试剂盒的组成及各种组分对保存期的影响, 对PCR和ELISA2种快速检测试剂盒进行比较。
结果该试剂盒能检测出20pg的沙门菌DNA; 在含有Taq酶、未冻干、-20℃的条件下, 试剂盒保存期为12个月; 以国标法做参照, ELISA试剂盒的敏感性和特异性分别为100%, 97.2%, PCR试剂盒的敏感性和特异性均为100%;ELISA和PCR试剂盒的符合率为97.2%;PCR试剂盒检测临床粪便样品150份, 检测结果与国标法相符。
结论沙门菌PCR试剂盒具有特异、灵敏、快速等优点, 适用于沙门菌快速检测。

Abstract:
Objective To develop a polymerase chain reaction(PCR)diag nostic kit for rapid detection of salmonella.
Methods Based on the effects of PCR detection, it was determined the components of PCR kit, and evaluated their affects on storage-life of kit.Meanwhile, the accuracy of both ELISA-kit and PCR-kit was compared.
Results The detection threshold of PCR kit was determined as 20 pg of salmonella DNA template.The storage-life of PCR-kit containing Taq DNA polymerase is longer than 12 months when it was stored at -20℃.The sensitivity and specificity of PCR method reached both 100%, while that of direct ELISA was 100% and 97.2%, respectively.The coincidence of these two methods reached 97.2%.150 human feces samples were detected by the PCR-kit and the results was coincided well with that of national standard(GB)method.
Conclusion PCR-kit was highly specific, sensitive and quick, it would be very useful in the rapid detection of salmonella.

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