Using propidium monoazide to distinguish between viable and nonviable bacteria, MS2 and murine norovirus
Using propidium monoazide to distinguish between viable and nonviable bacteria, MS2 and murine norovirus
复制标题
DOI：
10.1111/j.1472-765x.2012.03276.x
复制
发表时间：
2012-09-01
影响因子：
2.4
通讯作者：
Ko, G.
中科院分区：
文献类型：
--
作者：
Kim, S. Y.;Ko, G.
目的：区分活的和/或感染性微生物和灭活细胞的能力对于正确进行微生物风险评估极其重要。在这项研究中，我们评估了单叠氮丙啶（PMA）-qPCR是否可以区分活的和非活的细菌和病毒。方法和结果：将PMA-qPCR联合试验应用于活菌和灭活菌（大肠埃希菌和枯草芽孢杆菌）和病毒（MS 2和鼠诺如病毒[MNV]）。PMA是一种DNA嵌入剂，与PCR相结合能够更好地区分活的和非活的细菌和病毒比常规PCR。结论：这些结果表明，组合的PMA-qPCR测定可用于测量细菌细胞和噬菌体MS 2的活力，但不能测量MNV。该研究的意义和影响：PMA-qPCR可能用于测量某些微生物（包括病毒）的活力。但是，在通过PMA-qPCR定量测量微生物活力之前，应进行全面评价。
Aims: 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. Methods and Results: 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. Conclusions: 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. Significance and Impact of the Study: 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.