金黄色葡萄球菌活的非可培养状态复苏及PMA-qPCR检测.doc


金黄色葡萄球菌活的非可培养状态复苏及
PMA-qPCR 检测#
田聪,余以刚,肖性龙,吴晖**
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(华南理工大学轻工与食品学院,广州 510640)
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
中图分类号:;Q7
Resuscitation of viable but non-culturable state
us aureus and PMA-qPCR detection
TIAN Cong, YU Yigang, XIAO Xinglong, WU Hui
(College of Light Industry and Food Science, South China University of Technology,
GuangZhou 510640)
Abstract: In order to study the Resuscitation of viable but non-culturable state us
aureus, three factors: temperature, salt concentrations and pH posite to induce VBNC
us aureus. Four different resuscitations were attempted, qPCR and PMA in
combination with qPCR was established to detect ocus aureus. The date shows that the
VBNC samples could resuscitate in 48 h after disposed 8%Tween80. At the same time,
PMA-qPCR was a effective method for detection of VBNC ocus aureus, and could
e the ings of plate counts.
Keywords: ocus aureus; viable but nonculturable state; resuscitation; PMA; qPCR
0 引言
微生物在自然环境中以多种生理状态存在:可培养的活菌,活的非可培养状态,具有完
整结构但无生物活性的死菌以及细胞膜损伤死菌。徐怀恕等[1]通过对霍乱弧菌和大肠杆菌存
活规律的研究,发现细菌存在一种“活的非可培养状态”(viable but nonculturable state,
VBNC)。当细菌处于不良环境条件下时,用常规方法培养不能生