U.S. Environmental Protection Agency, National Exposure Research Laboratory, 26W. Martin Luther King Dr., Cincinnati, OH 45268, USA.
U.S. Environmental Protection Agency, National Exposure Research Laboratory, 26W. Martin Luther King Dr., Cincinnati, OH 45268, USA.
J Virol Methods. 2014 Apr;199:76-85. doi: 10.1016/j.jviromet.2014.01.005. Epub 2014 Jan 22.
The objective of this study was to compare three nucleic acid extraction and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) approaches for norovirus (NoV) detection in drinking water with respect to performance, costs, and analysis time. The approaches evaluated were: (A) an approach that utilizes the QIAamp DNA Blood Mini Kit and multiplex primers and probes for detection; (B) a procedure which includes the NucliSENS Magnetic Extraction Kit and other components of a proposed European Union standard method for NoV detection in foods; and (C) a commercialized assay which uses NucliSENS extraction and Cepheid SmartCycler® technologies. Each approach was evaluated by most probable number (MPN) analysis for detection of GI.1 and GII.4 NoVs from human stool. Furthermore, recoveries of spiked primary effluent in tap water concentrates were compared for each approach. Few significant differences were observed between approaches with regard to performance. However, Approach C was the most time consuming and expensive to perform. This research presents a case study of how molecular-based approaches for detection of NoVs can be compared and how various factors may play a role in which approach laboratories choose to employ.
本研究旨在比较三种核酸提取和逆转录定量聚合酶链反应 (RT-qPCR) 方法在饮用水中诺如病毒 (NoV) 的检测性能、成本和分析时间。评估的方法有：(A) 一种利用 QIAamp DNA 血液迷你试剂盒和多重引物和探针进行检测的方法；(B) 一种包括 NucliSENS 磁提取试剂盒和拟议的欧盟食品中 NoV 检测标准方法的其他成分的程序；(C) 一种商业化的测定法，使用 NucliSENS 提取和 Cepheid SmartCycler®技术。每种方法均通过最可能数 (MPN) 分析对从人粪便中提取的 GI.1 和 GII.4 NoVs 进行检测。此外，还比较了每种方法对自来水中加标原水的回收率。在性能方面，三种方法之间几乎没有明显差异。然而，方法 C 的耗时和费用最高。本研究提供了一个案例研究，说明如何比较基于分子的 NoV 检测方法，以及各种因素如何在实验室选择使用哪种方法方面发挥作用。
