Comparison of virus concentration methods for the RT-qPCR-based recovery of murine hepatitis virus, a surrogate for SARS-CoV-2 from untreated wastewater
Comparison of virus concentration methods for the RT-qPCR-based recovery of murine hepatitis virus, a surrogate for SARS-CoV-2 from untreated wastewater
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DOI：
10.1016/j.scitotenv.2020.139960
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发表时间：
2020-10-15
影响因子：
9.8
通讯作者：
Kitajima, Masaaki
中科院分区：
文献类型：
--
作者：
Ahmed, Warish;Bertsch, Paul M.;Kitajima, Masaaki
目前，许多国家将基于废水的流行病学（WBE）作为管理2019年冠状病毒病（COVID-19）全球大流行的现行措施的一部分，明显受益。由于大多数废水病毒浓缩方法都是针对非包膜病毒开发和验证的，因此有必要确定最常用的方法对包膜严重急性呼吸综合征冠状病毒2 （SARS-CoV-2）的效率。以人类冠状病毒（CoV）代剂小鼠肝炎病毒（MHV）为种子，对7种污水病毒浓缩方法（a - c）采用三种不同预处理方案的吸附萃取法、（D-E）采用两种不同装置的离心过滤装置法、（F)聚乙二醇（PEG 8000）沉淀法和(G)超离心法）的效率进行了测试。采用逆转录定量聚合酶链反应对MHV进行定量，并计算各方法的回收率。平均MHV回收率为26.7% ~ 65.7%。最有效的方法是有MgCl2预处理的吸附萃取法（方法C）和不经预处理的吸附萃取法（方法B）。第三种最有效的方法是Amicon (R) Ultra-15离心过滤装置（方法D），其回收率与最有效的方法无统计学差异。回收率最差的方法为酸化吸附萃取法(A)，其次为聚乙二醇沉淀法(F)。我们的研究结果表明，少量或不进行预处理的吸收-萃取方法可以提供适当的快速，成本效益和相对直接的废水中包膜病毒的回收。MHV是一种很有前景的SARS-CoV-2监测过程控制，可作为一种质量控制措施，支持社区层面的疫情缓解和风险评估。爱思唯尔B.V.版权所有
There is currently a clear benefit for many countries to utilize wastewater-based epidemiology (WBE) as part of ongoing measures to manage the coronavirus disease 2019 (COVID-19) global pandemic. Since most wastewater virus concentration methods were developed and validated for nonenveloped viruses, it is imperative to determine the efficiency of the most commonly used methods for the enveloped severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Municipal wastewater seeded with a human coronavirus (CoV) surrogate, murine hepatitis virus (MHV), was used to test the efficiency of seven wastewater virus concentration methods: (A-C) adsorption-extraction with three different pre-treatment options, (D-E) centrifugal filter device methods with two different devices, (F) polyethylene glycol (PEG 8000) precipitation, and (G) ultracentrifugation. MHV was quantified by reverse-transcription quantitative polymerase chain reaction and the recovery efficiency was calculated for each method. The mean MHV recoveries ranged from 26.7 to 65.7%. The most efficient methods were adsorption-extraction methods with MgCl2 pre-treatment (Method C), and without pre-treatment (Method B). The third most efficient method used the Amicon (R) Ultra-15 centrifugal filter device (Method D) and its recovery efficiency was not statistically different from the most efficient methods. The methods with the worst recovery efficiency included the adsorption-extraction method with acidification (A). followed by PEG precipitation (F). Our results suggest that absorption-extraction methods with minimal or without pre-treatment can provide suitably rapid, cost-effective and relatively straightforward recovery of enveloped viruses in wastewater. The MHV is a promising process control for SARS-CoV-2 surveillance and can be used as a quality control measure to support community-level epidemic mitigation and risk assessment. Crown Copyright (C) 2020 Published by Elsevier B.V.