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Journal article
Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR
Methods and protocols, Vol.4(1), 17
2021
PMID: 33672247
Abstract
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 900-1500x 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.
Details
- Title
- Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR
- Authors/Creators
- Kata Farkas - Bangor UniversityLuke S. Hillary - Bangor UniversityJamie Thorpe - Bangor UniversityDavid I. Walker - Ctr Environm Fisheries & Aquaculture Sci, UK Natl Reference Lab Foodborne Viruses, Weymouth DT4 8UB, EnglandJames A. Lowther - Centre for Environment, Fisheries and Aquaculture ScienceJames E. McDonald - Bangor UniversityShelagh K. Malham - Bangor UniversityDavey L. Jones - Bangor University
- Publication Details
- Methods and protocols, Vol.4(1), 17
- Publisher
- MDPI
- Number of pages
- 9
- Grant note
- 1937811 / NERC; UK Research & Innovation (UKRI); Natural Environment Research Council (NERC) NE/V004883/1 / UK Natural Environment Research Council (NERC); UK Research & Innovation (UKRI); Natural Environment Research Council (NERC) Soils Training and Research Studentship (STARS) grant from the Biotechnology and Biological Sciences Research Council (BBSRC); UK Research & Innovation (UKRI); Biotechnology and Biological Sciences Research Council (BBSRC) Defra Animal and Plant Health and Welfare (APHW) Official Development Assistance (ODA) COVID-19 Response Project NE/M009106/1 / Natural Environment Research Council; UK Research & Innovation (UKRI); Natural Environment Research Council (NERC)
- Identifiers
- 991005560420207891
- Copyright
- © 2021 by the authors.
- Murdoch Affiliation
- Centre for Sustainable Farming Systems
- Language
- English
- Resource Type
- Journal article
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