食品中金黄色葡萄球菌的荧光定量PCR检测法

Fluorescence Quantitative PCR (FQ-PCR) for Detection of Staphylococcus Aureus (SA) in Foods

目的建立一种简单、快速、准确、特异性强的能定量检测金黄色葡萄球菌的方法。方法以金黄色葡萄球菌FomB基因为靶序列,设计并合成引物和Taqman探针,优化引物与探针比例,调整镁离子浓度,对金黄色葡萄球菌进行荧光定量检测。结果引物与Taqman探针比例为1∶4,镁离子为2.5mmol/L时本底最低,荧光信号最强;该法的灵敏度1.0×103拷贝,能特异区分金色葡萄球菌与其他葡萄球菌。结论使用Taqman探针技术的荧光定量聚合酶链反应能够对金黄色葡萄球菌进行准确快速及定量检测。

[ Objective ] To establish a simple, rapid, accurate, speclhc and quantltat^ve metlaocl tor aetermlmng Stapnytoeoccus Aureus （SA）. [ Methods] The primer and Taqman probe were designed and synthesized with SA FemB gene as target sequence. The fluorescent quantitative detection was adopted to detect SA with optimizing the primer-probe ratio and regulating the eoneentration of Mg2^＋. [ Results] The lowest background and strongest fluorescence signal appeared when the primer-probe ratio was 1：4 and the eoncentration of Mg2^＋ was 2.5 mmol/L. The sensitivity of the method was 1.0 x 103 copies, and it could distinguish SA from other Staphylococcus. [ Conclusion ] The FQ-PCR based on taqman probes earl detect SA aecurately, rapidly and quantitatively.