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Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance
Citation:
Ahmed, W., S. Simpson, P. Bertsch, K. Bibby, A. Bivins, L. Blackall, S. Bofill-Mas, A. Bosch, J. Brandão, P. Choih, M. Ciesielski, E. Donner, N. D'Souza, A. Farnleitner, D. Gerrity, R. Gonzalez, J. Griffith, P. Gyawali, C. Haas, K. Hamilton, H. ChandithaHapuarachchi, V. Harwood, R. Haque, G. Jackson, S. Khan, W. Khan, M. Kitajima, A. Korajkic, G. La Rosa, B. Layton, E. Lipp, S. McLellan, B. McMinn, G. Medema, S. Metcalfe, W. Meijer, J. Mueller, H. Murphy, C. Naughton, R. Noble, S. Payyappat, S. Pettersonak, T. Pitkänenam, V. Rajal, B. Reyneke, F. Roman, J. Rose, M. Rusiñol, M. Sadowsky, L. Sala-Comorera, Y. Setoh, S. Sherchan, K. Sirikanchana, W. Smith, AND J. Steele. Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance. SCIENCE OF THE TOTAL ENVIRONMENT. Elsevier BV, AMSTERDAM, Netherlands, 805:149877, (2022). https://doi.org/10.1016/j.scitotenv.2021.149877
Impact/Purpose:
This manuscript is a comprehensive literature review detailing sources of false positive and false negative detection rates of SARS-CoV-2.
Description:
Wastewater surveillance for pathogens using reverse transcription-polymerase chain reaction (RT-PCR) is an effective and resource-efficient tool for gathering community-level public health information, including the incidence of coronavirus disease-19 (COVID-19). Surveillance of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) in wastewater can potentially provide an early warning signal of COVID-19 infections in a community. The capacity of the world's environmental microbiology and virology laboratories for SARS-CoV-2 RNA characterization in wastewater is increasing rapidly. However, there are no standardized protocols or harmonized quality assurance and quality control (QA/QC) procedures for SARS-CoV-2 wastewater surveillance. This paper is a technical review of factors that can cause false-positive and false-negative errors in the surveillance of SARS-CoV-2 RNA in wastewater, culminating in recommended strategies that can be implemented to identify and mitigate some of these errors. Recommendations include stringent QA/QC measures, representative sampling approaches, effective virus concentration and efficient RNA extraction, PCR inhibition assessment, inclusion of sample processing controls, and considerations for RT-PCR assay selection and data interpretation. Clear data interpretation guidelines (e.g., determination of positive and negative samples) are critical, particularly when the incidence of SARS-CoV-2 in wastewater is low. Corrective and confirmatory actions must be in place for inconclusive results or results diverging from current trends (e.g., initial onset or reemergence of COVID-19 in a community). It is also prudent to perform interlaboratory comparisons to ensure results' reliability and interpretability for prospective and retrospective analyses. The strategies that are recommended in this review aim to improve SARS-CoV-2 characterization and detection for wastewater surveillance applications. A silver lining of the COVID-19 pandemic is that the efficacy of wastewater surveillance continues to be demonstrated during this global crisis. In the future, wastewater should also play an important role in the surveillance of a range of other communicable diseases.
URLs/Downloads:
DOI: Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8386095/