Variability in RT-qPCR assay parameters indicates unreliable SARS-CoV-2 RNA quantification for wastewater surveillance

Bivins, Aaron and Kaya, Devrim and Bibby, Kyle and Simpson, Stuart L. and Bustin, Stephen A. and Shanks, Orin C. and Ahmed, Warish (2021) Variability in RT-qPCR assay parameters indicates unreliable SARS-CoV-2 RNA quantification for wastewater surveillance. Water Research, 203. p. 117516. ISSN 1879-2448

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Official URL: https://doi.org/10.1016/j.watres.2021.117516

Abstract

Due to the coronavirus disease 2019 (COVID-19) pandemic, wastewater surveillance has become an important tool for monitoring the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) within communities. In particular, reverse transcription-quantitative PCR (RT-qPCR) has been used to generate large datasets aimed at detecting and quantifying SARS-CoV-2 RNA in wastewater. Although RT-qPCR is rapid and sensitive, there is no standard method yet, there are no certified quantification standards, and experiments are conducted using different assays, reagents, instruments, and data analysis protocols. These variations can induce errors in quantitative data reports, thereby potentially misleading interpretations, and conclusions. We review the SARS-CoV-2 wastewater surveillance literature focusing on variability of RT-qPCR data as revealed by inconsistent standard curves and associated parameters. We find that variation in these parameters and deviations from best practices, as described in the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines suggest a frequent lack of reproducibility and reliability in quantitative measurements of SARS-CoV-2 RNA in wastewater.

Item Type: Journal Article
Keywords: RT-qPCR, assay validity, standard curve, quality assurance, quality control, wastewater surveillance
Faculty: Faculty of Science & Engineering
COVID-19 Research Collection
Depositing User: Lisa Blanshard
Date Deposited: 22 Sep 2021 11:13
Last Modified: 22 Sep 2021 11:16
URI: https://arro.anglia.ac.uk/id/eprint/706959

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