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 polymer...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-q PCR)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-log_(10) 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-log_(10) 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 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.展开更多
基金supported by a CHEO (Children’s Hospital of Eastern Ontario) CHAMO (Children’s Hospital Academic Medical Organization) grant, awarded to Dr.Alex E.Mac Kenziesupported by the “Next generation solutions to ensure healthy water resources for future generations” funded by the Global Water Futures program, Canada First Research Excellence Fund (#419205)+7 种基金supported by the Canada Research Chairs Program of the Natural Sciences and Engineering Research Council of Canada (NSERC)supported by funding from NSERC Discovery and Strategic Grant Programssupported by funding from an NSERC Discovery Grantsupported by the NSERC Alliance COVID-19 Grantby Mitacs through the Mitacs Accelerate programsupported by Canadian Institutes of Health Research (CIHR), Alberta Innovates, Alberta Health-Water for Life Strategysupported by the BC center for Disease Control, BC center for Disease Control Foundation for Public Health and Metro Vancouversupported by the Canada Research Chairs Program of NSERC。
文摘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-q PCR)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-log_(10) 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-log_(10) 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 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.
