Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Sci Total Environ. 2022 Jun 25;827:154258. doi: 10.1016/j.scitotenv.2022.154258. Epub 2022 Mar 3.
Waterborne diseases caused by pathogenic human viruses are a major public health concern. To control the potential risk of viral infection through contaminated waters, a rapid, reliable tool to assess the infectivity of pathogenic viruses is required. Recently, an advanced approach (i.e., capsid integrity (RT-)qPCR) was developed to discriminate intact viruses (potentially infectious) from inactivated viruses. In this approach, samples were pretreated with capsid integrity reagents (e.g., monoazide dyes or metal compounds) before (RT -)qPCR. These reagents can only penetrate inactivated viruses with compromised capsids to bind to viral genomes and prevent their amplification, but they cannot enter viruses with intact capsids. Therefore, only viral genomes of intact viruses were amplified or detected by (RT-)qPCR after capsid integrity treatment. In this study, we reviewed recent progress in the development and application of capsid integrity (RT-)qPCR to assess the potential infectivity of viruses (including non-enveloped and enveloped viruses with different genome structures [RNA and DNA]) in water. The efficiency of capsid integrity (RT-)qPCR has been shown to depend on various factors, such as conditions of integrity reagent treatment, types of viruses, environmental matrices, and the capsid structure of viruses after disinfection treatments (e.g., UV, heat, and chlorine). For the application of capsid integrity (RT-)qPCR in real-world samples, the use of suitable virus concentration methods and process controls is important to control the efficiency of capsid integrity (RT-)qPCR. In addition, potential future applications of capsid integrity (RT-)qPCR for determining the mechanism of disinfection treatment on viral structure (e.g., capsid or genome) and a combination of capsid integrity treatment and next-generation sequencing (NGS) (capsid integrity NGS) for monitoring the community of intact pathogenic viruses in water are also discussed. This review provides essential information on the application of capsid integrity (RT-)qPCR as an efficient tool for monitoring the presence of pathogenic viruses with intact capsids in water.
水中致病人类病毒引起的疾病是一个主要的公共卫生关注点。为了控制通过受污染的水感染病毒的潜在风险，需要一种快速、可靠的工具来评估致病病毒的感染力。最近，开发了一种先进的方法（即衣壳完整性（RT-）qPCR）来区分完整病毒（潜在感染性）和失活病毒。在这种方法中，样品在用衣壳完整性试剂（例如单偶氮染料或金属化合物）预处理后进行（RT-）qPCR。这些试剂只能穿透衣壳受损的失活病毒与病毒基因组结合并阻止其扩增，但不能进入衣壳完整的病毒。因此，只有经过衣壳完整性处理后，完整衣壳的病毒基因组才能通过（RT-）qPCR 扩增或检测到。在这项研究中，我们综述了衣壳完整性（RT-）qPCR 用于评估水中潜在感染性病毒（包括具有不同基因组结构[RNA 和 DNA]的非包膜和包膜病毒）的开发和应用的最新进展。衣壳完整性（RT-）qPCR 的效率已被证明取决于各种因素，例如完整性试剂处理条件、病毒类型、环境基质以及消毒处理后病毒的衣壳结构（例如 UV、热和氯）。为了在实际样品中应用衣壳完整性（RT-）qPCR，使用合适的病毒浓缩方法和过程控制对于控制衣壳完整性（RT-）qPCR 的效率非常重要。此外，衣壳完整性（RT-）qPCR 用于确定消毒处理对病毒结构（例如衣壳或基因组）的作用机制以及衣壳完整性处理与下一代测序（NGS）的结合（衣壳完整性 NGS）用于监测水中完整的致病病毒群落的潜在未来应用也进行了讨论。本综述提供了关于衣壳完整性（RT-）qPCR 作为监测水中完整衣壳致病病毒存在的有效工具的应用的重要信息。
