Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR.
Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR.
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DOI：
10.3390/mps4010017
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发表时间：
2021-02-23
影响因子：
2.4
通讯作者：
Jones DL
中科院分区：
文献类型：
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作者：
Farkas K;Hillary LS;Thorpe J;Walker DI;Lowther JA;McDonald JE;Malham SK;Jones DL
基于废水的流行病学已成为监视SARS-COV-2爆发的重要工具，但是，污水中的病毒发现是挑战，迄今为止，尚无对SARS-敏感检测的验证的标准方法COV-2在本文中，我们描述了一种基于聚乙二醇（PEG）的简单浓度方法（QRT-PCR）用于废水中的病毒检测。 –1500倍的废水样品浓度并充分消除了大多数有机物，这可以抑制随后的QRT-PCR分析。废水样品的物理化学特性，我们建议使用过程控制病毒来确定每个步骤的效率。对社区中病毒疾病进行全面评估的目标。
Wastewater-based epidemiology has become an important tool for the surveillance of SARS-CoV-2 outbreaks. However, the detection of viruses in sewage is challenging and to date there is no standard method available which has been validated for the sensitive detection of SARS-CoV-2. In this paper, we describe a simple concentration method based on polyethylene glycol (PEG) precipitation, followed by RNA extraction and a one-step quantitative reverse transcription PCR (qRT-PCR) for viral detection in wastewater. PEG-based concentration of viruses is a simple procedure which is not limited by the availability of expensive equipment and has reduced risk of disruption to consumable supply chains. The concentration and RNA extraction steps enable 900–1500× concentration of wastewater samples and sufficiently eliminates the majority of organic matter, which could inhibit the subsequent qRT-PCR assay. Due to the high variation in the physico-chemical properties of wastewater samples, we recommend the use of process control viruses to determine the efficiency of each step. This procedure enables the concentration and the extraction the DNA/RNA of different viruses and hence can be used for the surveillance of different viral targets for the comprehensive assessment of viral diseases in a community.