Taqman探针实时PCR检测金黄色葡萄球菌的研究  

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

作　　者：李一松[1,2] 韩希妍[1,2] 赵凤[1,2] 曲妍妍[1,2] 周艳秋[1,2] 李彬[1,2] 姜毓君[1,2] 

机构地区：[1]东北农业大学食品学院,乳品科学教育部重点实验室,黑龙江哈尔滨150030 [2]国家乳业工程技术研究中心,黑龙江哈尔滨150086

出　　处：《食品与发酵工业》2008年第5期156-161,共6页Food and Fermentation Industries

基　　金：国家科技支撑计划(2006BAD04A08-13);黑龙江省科技攻关重大项目(GA06C101-08);黑龙江省高校青年学术骨干项目(1151G006)

摘　　要：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。The presence of Staphylococcus aureus in milk is a major concern in the dairy industry. Staphylococcal 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 detection of S. aureus which harboring sea genes in raw milk.

关 键 词：TAQMAN 实时PCR 牛乳 金黄色葡萄球菌 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]