Comparison of approaches to quantify SARS-CoV-2 in wastewater using RT-qPCR: Results and implications from a collaborative inter-laboratory study in Canada
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 charact...
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| Veröffentlicht in: | Journal of environmental sciences (China) 2021-09, Vol.107, p.218-229 |
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| creator | 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 Conant, Bernadette Hrudey, Steve E. |
| description | 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 ( |
| doi_str_mv | 10.1016/j.jes.2021.01.029 |
| format | Article |
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[Display omitted]</description><identifier>ISSN: 1001-0742</identifier><identifier>EISSN: 1878-7320</identifier><identifier>EISSN: 1001-0742</identifier><identifier>DOI: 10.1016/j.jes.2021.01.029</identifier><identifier>PMID: 34412784</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>COVID-19 ; Humans ; Laboratories ; Pandemics ; Public health ; Quality assurance ; Quality control ; RNA, Viral ; SARS-CoV-2 ; Wastewater ; Wastewater surveillance</subject><ispartof>Journal of environmental sciences (China), 2021-09, Vol.107, p.218-229</ispartof><rights>2021</rights><rights>Copyright © 2021. Published by Elsevier B.V.</rights><rights>2021 The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. 2021</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c517t-40f483c8daa33d0774db50c93f8b86b0455788e8528b8ac06b143b81f8cea8933</citedby><cites>FETCH-LOGICAL-c517t-40f483c8daa33d0774db50c93f8b86b0455788e8528b8ac06b143b81f8cea8933</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jes.2021.01.029$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,780,784,885,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34412784$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chik, Alex H.S.</creatorcontrib><creatorcontrib>Glier, Melissa B.</creatorcontrib><creatorcontrib>Servos, Mark</creatorcontrib><creatorcontrib>Mangat, Chand S.</creatorcontrib><creatorcontrib>Pang, Xiao-Li</creatorcontrib><creatorcontrib>Qiu, Yuanyuan</creatorcontrib><creatorcontrib>D'Aoust, Patrick M.</creatorcontrib><creatorcontrib>Burnet, Jean-Baptiste</creatorcontrib><creatorcontrib>Delatolla, Robert</creatorcontrib><creatorcontrib>Dorner, Sarah</creatorcontrib><creatorcontrib>Geng, Qiudi</creatorcontrib><creatorcontrib>Giesy, John P.</creatorcontrib><creatorcontrib>McKay, Robert Mike</creatorcontrib><creatorcontrib>Mulvey, Michael R.</creatorcontrib><creatorcontrib>Prystajecky, Natalie</creatorcontrib><creatorcontrib>Srikanthan, Nivetha</creatorcontrib><creatorcontrib>Xie, Yuwei</creatorcontrib><creatorcontrib>Conant, Bernadette</creatorcontrib><creatorcontrib>Hrudey, Steve E.</creatorcontrib><creatorcontrib>Canadian SARS-CoV-2 Inter-Laboratory Consortium</creatorcontrib><title>Comparison of approaches to quantify SARS-CoV-2 in wastewater using RT-qPCR: Results and implications from a collaborative inter-laboratory study in Canada</title><title>Journal of environmental sciences (China)</title><addtitle>J Environ Sci (China)</addtitle><description>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 (<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.
[Display omitted]</description><subject>COVID-19</subject><subject>Humans</subject><subject>Laboratories</subject><subject>Pandemics</subject><subject>Public health</subject><subject>Quality assurance</subject><subject>Quality control</subject><subject>RNA, Viral</subject><subject>SARS-CoV-2</subject><subject>Wastewater</subject><subject>Wastewater surveillance</subject><issn>1001-0742</issn><issn>1878-7320</issn><issn>1001-0742</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kdtq3DAQhkVpaQ7tA_Sm6AW8kWR5JbdQCKYnCLRs0t6KsSQnWmzJkeQN-yx92WrZNLQ3hQGNZub_huFH6A0lK0ro-mK72tq0YoTRFSnB2mfolEohK1Ez8rzkhNCKCM5O0FlKW0IIb0jzEp3UnFMmJD9Fv7owzRBdCh6HAcM8xwD6ziacA75fwGc37PH15ea66sLPimHn8QOkbB8g24iX5Pwt3txU99-7zTu8sWkZc8LgDXbTPDoN2QWf8BDDhAHrMI7Qh1iqO1tQBVE9FkLc45QXsz9s6MCDgVfoxQBjsq8f33P049PHm-5LdfXt89fu8qrSDRW54mTgstbSANS1IUJw0zdEt_Uge7nuy82NkNLKhpU_aLLuKa97SQepLci2rs_RhyN3XvrJGm19jjCqOboJ4l4FcOrfjnd36jbslGhZK-QBQI8AHUNK0Q5PWkrUwSm1VcUpdXBKkRKsLZq3fy99Uvyxpgy8Pw7YcvrO2aiSdtZra1y0OisT3H_wvwH5VKe9</recordid><startdate>20210901</startdate><enddate>20210901</enddate><creator>Chik, Alex H.S.</creator><creator>Glier, Melissa B.</creator><creator>Servos, Mark</creator><creator>Mangat, Chand S.</creator><creator>Pang, Xiao-Li</creator><creator>Qiu, Yuanyuan</creator><creator>D'Aoust, Patrick M.</creator><creator>Burnet, Jean-Baptiste</creator><creator>Delatolla, Robert</creator><creator>Dorner, Sarah</creator><creator>Geng, Qiudi</creator><creator>Giesy, John P.</creator><creator>McKay, Robert Mike</creator><creator>Mulvey, Michael R.</creator><creator>Prystajecky, Natalie</creator><creator>Srikanthan, Nivetha</creator><creator>Xie, Yuwei</creator><creator>Conant, Bernadette</creator><creator>Hrudey, Steve E.</creator><general>Elsevier B.V</general><general>The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences. 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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 (<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.
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| fulltext | fulltext |
| identifier | ISSN: 1001-0742 |
| ispartof | Journal of environmental sciences (China), 2021-09, Vol.107, p.218-229 |
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| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7929783 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present); Alma/SFX Local Collection |
| subjects | COVID-19 Humans Laboratories Pandemics Public health Quality assurance Quality control RNA, Viral SARS-CoV-2 Wastewater Wastewater surveillance |
| title | Comparison of approaches to quantify SARS-CoV-2 in wastewater using RT-qPCR: Results and implications from a collaborative inter-laboratory study in Canada |
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