Taqman探针实时PCR检测金黄色葡萄球菌的研究

建立了Taqman探针实时PCR方法,针对乳中携带sea基因的金黄色葡萄球菌进行检测。研究所设计的引物具有良好的特异性,而且Taqman探针实时PCR方法检测sea基因的灵敏度高,最低检出限为69 fg,并且该方法可在8 h内完成人工污染乳中金黄色葡萄球菌的检测,最低检出限为83 cfu/mL。研究所建立的方法具有特异性强、灵敏性高及操作简便等优点,适宜于牛乳中金黄色葡萄球菌污染的调查及监测。

Study on Quantification of Staphylococcus aureus in Milk by Real-time Taqman PCR

The presence of Staphylococcus aureus in milk is a major concern in the dairy industry.Staphy- lococcal Enterotoxin A(SEA)is among the most potent of the growing list of known enterotoxins produced by S.aureus.A dose of less than 1μg of enterotoxins in contaminated food will produce symptoms after 1- 6 h in sensitive persons(FDA reported).Outbreaks as well as sporadic cases involving contaminated milk and dairy products have been reported.In this study,a real-time PCR assays,using dye-labeled DNA probes, was developed to identify the S.aureus which harboring sea genes.The detection procedure was applied to sterile milk samples that were artificially contaminated by S.aureus.The experimental result indicated the primers design had the good specificity and the detection limit was 69fg,and the developed assay enables us to detect 83 cfu/mL in sterile milk.The assay can be accomplished within 8 hours.This work can really offer a very quick,reliable alternative to identify the S.aureus.And it would offer great help to the further detec- tion of S.aureus which harboring sea genes in raw milk.