1. Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance
- Author
-
Ahmed, W, Simpson, SL, Bertsch, PM, Bibby, K, Bivins, A, Blackall, LL, Bofill-Mas, S, Bosch, A, Brandao, J, Choi, PM, Ciesielski, M, Donner, E, D'Souza, N, Farnleitner, AH, Gerrity, D, Gonzalez, R, Griffith, JF, Gyawali, P, Haas, CN, Hamilton, KA, Hapuarachchi, HC, Harwood, VJ, Haque, R, Jackson, G, Khan, SJ, Khan, W, Kitajima, M, Korajkic, A, La Rosa, G, Layton, BA, Lipp, E, McLellan, SL, McMinn, B, Medema, G, Metcalfe, S, Meijer, WG, Mueller, JF, Murphy, H, Naughton, CC, Noble, RT, Payyappat, S, Petterson, S, Pitkanen, T, Rajal, VB, Reyneke, B, Roman, FA, Rose, JB, Rusinol, M, Sadowsky, MJ, Sala-Comorera, L, Setoh, YX, Sherchan, SP, Sirikanchana, K, Smith, W, Steele, JA, Subburg, R, Symonds, EM, Thai, P, Thomas, K, Tynan, J, Toze, S, Thompson, J, Whiteley, AS, Wong, JCC, Sano, D, Wuertz, S, Xagoraraki, I, Zhang, Q, Zimmer-Faust, AG, Shanks, OC, Ahmed, W, Simpson, SL, Bertsch, PM, Bibby, K, Bivins, A, Blackall, LL, Bofill-Mas, S, Bosch, A, Brandao, J, Choi, PM, Ciesielski, M, Donner, E, D'Souza, N, Farnleitner, AH, Gerrity, D, Gonzalez, R, Griffith, JF, Gyawali, P, Haas, CN, Hamilton, KA, Hapuarachchi, HC, Harwood, VJ, Haque, R, Jackson, G, Khan, SJ, Khan, W, Kitajima, M, Korajkic, A, La Rosa, G, Layton, BA, Lipp, E, McLellan, SL, McMinn, B, Medema, G, Metcalfe, S, Meijer, WG, Mueller, JF, Murphy, H, Naughton, CC, Noble, RT, Payyappat, S, Petterson, S, Pitkanen, T, Rajal, VB, Reyneke, B, Roman, FA, Rose, JB, Rusinol, M, Sadowsky, MJ, Sala-Comorera, L, Setoh, YX, Sherchan, SP, Sirikanchana, K, Smith, W, Steele, JA, Subburg, R, Symonds, EM, Thai, P, Thomas, K, Tynan, J, Toze, S, Thompson, J, Whiteley, AS, Wong, JCC, Sano, D, Wuertz, S, Xagoraraki, I, Zhang, Q, Zimmer-Faust, AG, and Shanks, OC
- Abstract
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 pandemi
- Published
- 2022