Concentration and quantification of SARS-CoV-2 RNA in wastewater using polyethylene glycol-based concentration and qRT-PCR

Kata Farkas*, Luke Hillary, Jamie Thorpe, David Walker, James A. Lowther, James McDonald, Shelagh Malham, Davey L. Jones

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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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 900-1500× 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.
Original languageEnglish
Article number17
Number of pages9
JournalMethods and Protocols
Volume4
Issue number1
DOIs
Publication statusPublished - 23 Feb 2021

Keywords

  • COVID-19
  • public health
  • wastewater virology
  • Sewage
  • environmental sample

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