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

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posted on 2023-07-26, 15:31 authored by Aaron Bivins, Devrim Kaya, Kyle Bibby, Stuart L. Simpson, Stephen A. Bustin, Orin C. Shanks, Warish Ahmed
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.

History

Refereed

  • Yes

Volume

203

Page range

117516

Publication title

Water Research

ISSN

1879-2448

Publisher

Elsevier

File version

  • Published version

Language

  • eng

Legacy posted date

2021-09-22

Legacy creation date

2021-09-22

Legacy Faculty/School/Department

Faculty of Science & Engineering

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