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

• 作者: Alex H.S.Chik Melissa B.Glier Mark Servos Chand S.Mangat Xiao-Li Pang Yuanyuan Qiu Patrick M.D'Aoust Jean-Baptiste Burnet Robert Delatolla Sarah Dorner Qiudi Geng John P.Giesy Jr Robert Mike McKay Michael R.Mulvey Natalie Prystajecky Nivetha Srikanthan Yuwei Xie Bernadette Conant Steve E.Hrudey Canadian SARS-CoV- Inter-Laboratory Consortium
• 来源期刊: 环境科学学报（英文版）
• 年，卷（期）: 2021,（9）
• 所属期刊栏目: Regular articles
• 页码范围: 218-229
• 页数: 12页
• 语种: 英文

摘要

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 re-verse transcription quantitative polymerase chain reaction (RT-qPCR) are still far from stan-dardized 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 andhuman coronavirus strain 229E [HCoV-229E]) at low and high levels then provided "blind" to eight laboratories. Concentration estimates reported by individual laboratories were con-sistently 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 vari-ability 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 (<20 gene copies/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 repre-sent 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 con-sidered to evaluate method effectiveness. A consistent method and laboratory to explore wastewater SARS-CoV-2 temporal trends for a given system, with appropriate quality con-trol protocols and documented in adequate detail should succeed.