Comparison of approaches to quantify SARS-CoV-2 in wastewater using RT-qPCR: Results and implications from a collaborative inter-laboratory study in Canada

Authors

Alex H.S. Chik, Consultant to Canadian Water Network Inc.
Melissa B. Glier, BC Centre for Disease Control
Mark Servos, University of Waterloo
Chand S. Mangat, National Microbiology Laboratory
Xiao Li Pang, Alberta Precision Laboratories
Yuanyuan Qiu, University of Alberta
Patrick M. D'Aoust, University of Ottawa
Jean Baptiste Burnet, Polytechnique Montréal
Robert Delatolla, University of Ottawa
Sarah Dorner, Polytechnique Montréal
Qiudi Geng, Great Lakes Institute for Environmental Research
John P. Giesy, University of Saskatchewan
Robert Mike McKay, Great Lakes Institute for Environmental Research
Michael R. Mulvey, National Microbiology Laboratory
Natalie Prystajecky, BC Centre for Disease Control
Nivetha Srikanthan, University of Waterloo
Yuwei Xie, University of Saskatchewan
Bernadette Conant, Canadian Water Network

Document Type

Article

Publication Date

9-1-2021

Publication Title

Journal of Environmental Sciences (China)

Volume

107

First Page

218

Keywords

COVID-19, Public health, Quality assurance, Quality control, Wastewater surveillance

Last Page

229

Abstract

Detection of SARS-CoV-2 RNA in wastewater is a promising tool for informing public health decisions during the COVID-19 pandemic. However, approaches for its analysis by use of reverse transcription quantitative polymerase chain reaction (RT-qPCR) are still far from standardized globally. To characterize inter- and intra-laboratory variability among results when using various methods deployed across Canada, aliquots from a real wastewater sample were spiked with surrogates of SARS-CoV-2 (gamma-radiation inactivated SARS-CoV-2 and human coronavirus strain 229E [HCoV-229E]) at low and high levels then provided “blind” to eight laboratories. Concentration estimates reported by individual laboratories were consistently within a 1.0-log10 range for aliquots of the same spiked condition. All laboratories distinguished between low- and high-spikes for both surrogates. As expected, greater variability was observed in the results amongst laboratories than within individual laboratories, but SARS-CoV-2 RNA concentration estimates for each spiked condition remained mostly within 1.0-log10 ranges. The no-spike wastewater aliquots provided yielded non-detects or trace levels (/mL) of SARS-CoV-2 RNA. Detections appear linked to methods that included or focused on the solids fraction of the wastewater matrix and might represent in-situ SARS-CoV-2 to the wastewater sample. HCoV-229E RNA was not detected in the no-spike aliquots. Overall, all methods yielded comparable results at the conditions tested. Partitioning behavior of SARS-CoV-2 and spiked surrogates in wastewater should be considered to evaluate method effectiveness. A consistent method and laboratory to explore wastewater SARS-CoV-2 temporal trends for a given system, with appropriate quality control protocols and documented in adequate detail should succeed.

DOI

10.1016/j.jes.2021.01.029

ISSN

10010742

E-ISSN

18787320

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Chik, Alex H.S.; Glier, Melissa B.; Servos, Mark; Mangat, Chand S.; Pang, Xiao Li; Qiu, Yuanyuan; D'Aoust, Patrick M.; Burnet, Jean Baptiste; Delatolla, Robert; Dorner, Sarah; Geng, Qiudi; Giesy, John P.; McKay, Robert Mike; Mulvey, Michael R.; Prystajecky, Natalie; Srikanthan, Nivetha; Xie, Yuwei; and Conant, Bernadette. (2021). Comparison of approaches to quantify SARS-CoV-2 in wastewater using RT-qPCR: Results and implications from a collaborative inter-laboratory study in Canada. Journal of Environmental Sciences (China), 107, 218-229.
https://scholar.uwindsor.ca/glierpub/535

PubMed ID

34412784