牛奶中金黄色葡萄球菌PCR检测方法的建立与应用

Consturction and Application of a PCR-based Assay for Staphylococcus aureus in Milk

目的:建立一种直接从牛奶中检测金黄色葡萄球菌的PCR快速诊断方法。方法:根据已发表的金黄色葡萄球菌16-23SrRNA特异性序列设计并合成一对引物,通过优化反应条件建立从牛奶中直接检测金黄色葡萄球菌的PCR方法。结果:与细菌的常规分离方法相比,PCR法敏感性高,与其它型葡萄球菌无交叉反应,特异性达100%,检测细菌基因组DNA最低浓度为35ng/μL,能在5h内对送检的牛奶样品直接进行金黄色葡萄球菌的测定,可用于乳制品中金黄色葡萄球菌的检测。结论:成功建立了快速检测牛奶中金黄色葡萄球菌PCR方法。

Objective： To establish a rapid diagnostic polymerase chain reaction （PCR） assay for direct detection of bacterial pathogens in raw milk from cows with mastitis. Methods： The primers were designed based on the published gene sequences coding for 16-23S rRNA of Staphylococcus aureus （S.aureus）, and a PCR-based method was developed and optimized to identify S.aureus in milk, Results： The experimental results show that this PCR assay could detect S.aureus in milk samples within 5 hours, including extracting bacterial genome DNA. This PCR assay had virtues of good sensitivity, specificity and stability, and its sensitivity could be as little as 35 ng/izl of DNA. This method can be used for identification for bacterial pathogens in milk products. Conclusions A method was developed to detect S.aurens in milk by PCR.