Concentration and quantification of sars-cov-2 rna in wastewater using polyethylene glycol-based concentration and qrt-pcr

Kata Farkas, Luke S. Hillary, Jamie Thorpe, David I. Walker, James A. Lowther, James E. McDonald, Shelagh K. Malham, Davey L. Jones

Research output: Contribution to journalArticlepeer-review

10 Citations (Web of Science)

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 physicochemical 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
Pages (from-to)1-9
Number of pages9
JournalMethods and protocols
Volume4
Issue number1
DOIs
Publication statusPublished - 2021

Fingerprint

Dive into the research topics of 'Concentration and quantification of sars-cov-2 rna in wastewater using polyethylene glycol-based concentration and qrt-pcr'. Together they form a unique fingerprint.

Cite this