金黄色葡萄球菌活非可培养状态复苏和PMA-qPCR检测.PDFVIP

现代食品科技 Modern Food Science and Technology 2013, Vol.29, No.6 金黄色葡萄球菌活的非可培养状态复苏及 PMA-qPCR 检测 1 1 1 1 1 2 田聪 ，余以刚 ，肖性龙 ，吴晖 ，赖富饶 ，杨锡洪 （1.华南理工大学轻工与食品学院，广东广州 510640）（2.广东温氏佳润食品有限公司，广东云浮 527400） 摘要：为研究金黄色葡萄球菌活的非可培养（VBNC ）状态的复苏问题，采取温度、盐度和酸度3 个因素复合诱导制备VBNC 菌，尝试四种复苏方法，并以荧光定量PCR 和PMA-qPCR 技术结合的方法进行检测。结果证明：8%无菌Tween80 可以使VBNC 状 态金黄色葡萄球菌48 h 后复苏，同时PMA-qPCR 能够有效检出VBNC 状态金黄色葡萄球菌，克服传统平板计数法对于VBNC 菌的 漏检。 关键词：金黄色葡萄球菌；活的非可培养状态；复苏；PMA ；荧光定量PCR 文章篇号：1673-9078(2013)6-1390-1394 Resuscitation and PMA-qPCR Detection of Staphylococcus aureus in the Viable but Non-culturable State 1 1 1 1 1 2 TIAN Cong , YU Yi-gang , XIAO Xing-long , WU Hui , LAI Fu-rao , YANG Xi-hong (1.College of Light Industry and Food Science, South China University of Technology, Guangzhou 510640, China) (2.Guangdong Wen’s Caren Food Co., Ltd. ,Yunfu 527400, China) Abstract: In order to study the resuscitation of Staphylococcus aureus in the viable but non-culturable state (VBNC), three factors (temperature, salt concentrations and pH) were investigated to induce VBNC Staphylococcus aureus. Four different resuscitation methods were attempted and a new detection method using qPCR and PMA in combination with qPCR was established to detect Staphyloccocus aureus. The results showed that the VBNC samples could resuscitate in 48 h after disposed in 8% Tween80. In addition, PMA-qPCR was an effective method for detection of VBNC Staphyloccocus aureus, which efficiently overcome the shortcomings of plate counts. Key words: Staphyloccocus aureus ; viable but nonculturable state; resuscitation; PMA; qPCR 微生物在自然环境