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人类诺如病毒检测方法及其感染力预测的批判性评价。

A critical review of methods for detecting human noroviruses and predicting their infectivity.

机构信息

Leatherhead Food Research, Leatherhead, UK.

出版信息

Crit Rev Microbiol. 2013 Aug;39(3):295-309. doi: 10.3109/1040841X.2012.709820. Epub 2012 Aug 20.

Abstract

Human noroviruses (hNoVs) are the single most common cause of acute non-bacterial gastroenteritis in the industrialized world but cannot be grown in simple culture systems. Different approaches for detecting hNoVs and predicting their infectivity are reviewed. Although reverse transcription quantitative PCR (RT-qPCR) is the most widely used method to detect human noroviruses (hNoVs) it is unable to discriminate between infectious and non-infectious particles. There is therefore a dilemma in assessing the risk to human health from samples detected as positive in RT-qPCR assays. In the absence of an efficient cell, culture based detection system for hNoVs RT-qPCR methods need to differentiate RT-qPCR signals from intact infective particles, intact defective particles, degraded particles (consisting of capsid protein and virus RNA, herein referred to as ribonucleoprotein complexes (RNPs), and "naked" RNA. This review provides a critical analysis of methods for detecting hNoVs, and differentiating such signals with reference to relevant studies of virus infectivity, structure, inactivation, and the disassembly of virus particles during infection. The application of these methods as an adjunct to the proposed RT-qPCR European Committee for Standards (CEN) methods for the detection of hNoVs in foods and the environment is discussed.

摘要

人类诺如病毒(hNoVs)是工业化世界中引起急性非细菌性胃肠炎的单一最常见原因,但不能在简单的培养系统中生长。本文综述了用于检测 hNoVs 并预测其感染性的不同方法。尽管逆转录定量 PCR(RT-qPCR)是检测人类诺如病毒(hNoVs)最广泛使用的方法,但它无法区分传染性和非传染性颗粒。因此,在评估 RT-qPCR 检测呈阳性的样本对人类健康的风险时存在困境。在缺乏有效的基于细胞的 hNoVs 培养检测系统的情况下,RT-qPCR 方法需要区分 RT-qPCR 信号与完整的感染性颗粒、完整的缺陷颗粒、降解颗粒(由衣壳蛋白和病毒 RNA 组成,本文称为核糖核蛋白复合物(RNPs))和“裸露”RNA。本文批判性地分析了检测 hNoVs 的方法,并参考了有关病毒感染性、结构、失活以及感染过程中病毒颗粒解体的相关研究,对这些信号进行了区分。讨论了将这些方法应用于拟议的 RT-qPCR 欧洲标准委员会(CEN)方法,以检测食品和环境中的 hNoVs。

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