基于TaqMan探针的荧光定量PCR检测水产品中的沙门菌

Detection of Salmonella in seafoods by TaqMan-based real-time fluorescent quantitative PCR

目的:建立基于TaqMan探针的荧光定量PCR检测水产品中沙门菌的检测体系。方法:根据沙门菌外膜蛋白Ompc基因的保守序列,设计1对引物和TaqMan探针,并对荧光定量PCR反应体系和反应条件进行优化。利用建立的检测方法对舟山水产品加工厂和舟山菜市场采集的120份样本进行检测。结果:该检测方法可以达到1μl 1.0×101拷贝数量级的灵敏度。样品检出率为10.0%,与国标法检测结果一致。结论:本研究建立的TaqMan探针荧光定量PCR检测方法可以特异、灵敏、简便快速的实现对水产品中沙门菌的检测。

Objective：To develop the TaqMan-based real-time PCR assay for detection of salmonella in aquatic products.Methods： Primers and TaqMan probe were designed based on Ompc gene sequence of salmonella,and salmonella detection was achieved by optimizing Real-time PCR reaction condition and system.With TaqMan-based real-time PCR method,120 aquatic products were analyzed.Results： This method had high sensitivity and its detection limit was 10 copies per μl.And the result showed that the positive rate of salmonella contamination was 10.0%,which was consistent with the national standard approach.Conclusion： The TaqMan-based real-time PCR method is specific,sensitive,rapid and easy for detection of salmonella in aquatic products.