Protocol for safe, affordable, and reproducible isolation and quantitation of SARS-CoV-2 RNA from wastewater.
Protocol for safe, affordable, and reproducible isolation and quantitation of SARS-CoV-2 RNA from wastewater.
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DOI：
10.1371/journal.pone.0257454
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Dennehy JJ
中科院分区：
文献类型：
--
作者：
Trujillo M;Cheung K;Gao A;Hoxie I;Kannoly S;Kubota N;San KM;Smyth DS;Dennehy JJ
以下方案描述了我们处理废水的工作流程，目的是检测SARS-CoV-2的遗传信号。步骤包括巴氏杀菌、病毒浓缩、RNA提取和RT-qPCR定量。我们包括辅助步骤，为新用户提供工具和策略，以帮助解决流程中的关键步骤。该方案是检测废水中SARS-CoV-2 RNA的最安全、最便宜和最可复制的方法之一。由于巴氏灭菌步骤，它在BSL2设施中使用是安全的。除了使方案对相关人员安全之外，巴氏消毒还有增加SARS-CoV-2遗传信号的额外好处。此外，使用该方案获得的RNA可以使用Sanger和Illumina测序技术进行测序。该议定书由纽约市环境保护部于2020年8月通过，用于监测该市所有五个行政区废水中SARS-CoV-2的流行情况。在未来，该方案可用于检测废水中各种其他临床相关病毒，并作为监测已知和新出现的病毒病原体社区传播的废水监测策略的基础。
The following protocol describes our workflow for processing wastewater with the goal of detecting the genetic signal of SARS-CoV-2. The steps include pasteurization, virus concentration, RNA extraction, and quantification by RT-qPCR. We include auxiliary steps that provide new users with tools and strategies that will help troubleshoot key steps in the process. This protocol is one of the safest, cheapest, and most reproducible approaches for the detection of SARS-CoV-2 RNA in wastewater. Owing to a pasteurization step, it is safe for use in a BSL2 facility. In addition to making the protocol safe for the personnel involved, pasteurization had the added benefit of increasing the SARS-CoV-2 genetic signal. Furthermore, the RNA obtained using this protocol can be sequenced using both Sanger and Illumina sequencing technologies. The protocol was adopted by the New York City Department of Environmental Protection in August 2020 to monitor SARS-CoV-2 prevalence in wastewater in all five boroughs of the city. In the future, this protocol could be used to detect a variety of other clinically relevant viruses in wastewater and serve as a foundation of a wastewater surveillance strategy for monitoring community spread of known and emerging viral pathogens.
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DOI：
10.1016/j.coesh.2020.09.006
发表时间：
2020-10
影响因子：
--
作者：
Medema G;Been F;Heijnen L;Petterson S
通讯作者：
Petterson S
DOI：
10.1039/d0ew00946f
发表时间：
2021-01-20
期刊：
Environmental science : water research & technology
影响因子：
--
作者：
Pecson BM;Darby E;Haas CN;Amha YM;Bartolo M;Danielson R;Dearborn Y;Di Giovanni G;Ferguson C;Fevig S;Gaddis E;Gray D;Lukasik G;Mull B;Olivas L;Olivieri A;Qu Y;SARS-CoV-2 Interlaboratory Consortium
通讯作者：
SARS-CoV-2 Interlaboratory Consortium
影响因子：
6.4
作者：
Crits-Christoph A;Kantor RS;Olm MR;Whitney ON;Al-Shayeb B;Lou YC;Flamholz A;Kennedy LC;Greenwald H;Hinkle A;Hetzel J;Spitzer S;Koble J;Tan A;Hyde F;Schroth G;Kuersten S;Banfield JF;Nelson KL
通讯作者：
Nelson KL
影响因子：
5.4
作者：
VIDAVER, AK;KOSKI, RK;VANETTEN, JL
通讯作者：
VANETTEN, JL
DOI：
10.1016/j.scitotenv.2021.145274
发表时间：
2021-05-20
期刊：
The Science of the total environment
影响因子：
--
作者：
Hokajärvi AM;Rytkönen A;Tiwari A;Kauppinen A;Oikarinen S;Lehto KM;Kankaanpää A;Gunnar T;Al-Hello H;Blomqvist S;Miettinen IT;Savolainen-Kopra C;Pitkänen T
通讯作者：
Pitkänen T