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Correction published on 12 November 2021, see Methods Protoc. 2021, 4(4), 82.
Protocol

Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR

1
School of Natural Sciences, Bangor University, Deiniol Road, Bangor LL57 2UW, UK
2
School of Ocean Sciences, Bangor University, Menai Bridge LL59 5AB, UK
3
UK National Reference Laboratory for Foodborne Viruses, Centre for Environment, Fisheries and Aquaculture Science, Weymouth DT4 8UB, UK
4
UWA School of Agriculture and Environment, The University of Western Australia, Perth, WA 6009, Australia
*
Author to whom correspondence should be addressed.
Methods Protoc. 2021, 4(1), 17; https://doi.org/10.3390/mps4010017
Received: 21 January 2021 / Revised: 17 February 2021 / Accepted: 19 February 2021 / Published: 23 February 2021 / Corrected: 12 November 2021
(This article belongs to the Section Public Health Research)
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. View Full-Text
Keywords: COVID-19; environmental samples; sewage; wastewater virology; public health COVID-19; environmental samples; sewage; wastewater virology; public health
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MDPI and ACS Style

Farkas, K.; Hillary, L.S.; Thorpe, J.; Walker, D.I.; Lowther, J.A.; McDonald, J.E.; Malham, S.K.; Jones, D.L. Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR. Methods Protoc. 2021, 4, 17. https://doi.org/10.3390/mps4010017

AMA Style

Farkas K, Hillary LS, Thorpe J, Walker DI, Lowther JA, McDonald JE, Malham SK, Jones DL. Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR. Methods and Protocols. 2021; 4(1):17. https://doi.org/10.3390/mps4010017

Chicago/Turabian Style

Farkas, Kata, Luke S. Hillary, Jamie Thorpe, David I. Walker, James A. Lowther, James E. McDonald, Shelagh K. Malham, and Davey L. Jones 2021. "Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR" Methods and Protocols 4, no. 1: 17. https://doi.org/10.3390/mps4010017

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