Concentration and quantification of SARS-CoV-2 RNA in wastewater using polyethylene glycol-based concentration and qRT-PCR
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- mps-04-00017 (2)
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DOI
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.
Keywords
- COVID-19, public health, wastewater virology, Sewage, environmental sample
Original language | English |
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Article number | 17 |
Number of pages | 10 |
Journal | Methods and Protocols |
Volume | 4 |
Issue number | 1 |
DOIs | |
Publication status | Published - 23 Feb 2021 |