Department of Environmental Health, School of Public Health, Institute of Health and Environment, Seoul National University, Seoul, Korea.
Lett Appl Microbiol. 2012 Sep;55(3):182-8. doi: 10.1111/j.1472-765X.2012.03276.x. Epub 2012 Jul 20.
The ability to distinguish between viable and/or infectious micro-organisms and inactivated cells is extremely important for correctly performing microbial risk assessments. In this study, we evaluated whether propidium monoazide (PMA)-qPCR could distinguish between viable and nonviable bacteria and viruses.
A PMA-qPCR combined assay was applied to viable and inactivated bacteria (Escherichia coli and Bacillus subtilis) and viruses (MS2 and murine norovirus [MNV]). PMA, a DNA-intercalating agent, in combination with PCR was better able to distinguish between viable and nonviable bacteria and viruses than conventional PCR.
These results suggest that a combined PMA-qPCR assay can be used to measure the viability of bacterial cells and bacteriophage MS2, but not MNV.
PMA-qPCR could potentially be used to measure the viability of some micro-organisms, including virus. However, a thorough evaluation should be performed prior to measuring the viability of micro-organisms by PMA-qPCR in a quantitative way.
能够区分有活力和/或传染性的微生物与失活细胞对于正确进行微生物风险评估非常重要。在本研究中，我们评估了吖啶橙单加合物（PMA）-qPCR 是否能够区分有活力和无活力的细菌和病毒。
将 PMA-qPCR 联合检测应用于有活力和失活的细菌（大肠杆菌和枯草芽孢杆菌）和病毒（MS2 和鼠诺如病毒[MNV]）。吖啶橙是一种 DNA 嵌入剂，与 PCR 联合使用比传统 PCR 更能区分有活力和无活力的细菌和病毒。
这些结果表明，PMA-qPCR 联合检测可用于测量细菌细胞和噬菌体 MS2 的活力，但不能测量 MNV 的活力。
PMA-qPCR 可能可用于测量某些微生物的活力，包括病毒。但是，在通过 PMA-qPCR 以定量方式测量微生物的活力之前，应该进行彻底的评估。
