Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance

• Autores: Ahmed, Warish; Simpson, Stuart L.; Bertsch, Paul M.; Bibby, Kyle; Bivins, Aaron; Blackall, Linda L.; Bofill-Mas, Sílvia; Bosch, Albert; Brandão, João; Choi, Phil M.; Ciesielski, Mark; Donner, Erica; D'Souza, Nishita; Farnleitner, Andreas H.; Gerrity, Daniel; Gonzalez, Raul; Griffith, John F.; Gyawali, Pradip; Haas, Charles N.; Hamilton, Kerry A.; Hapuarachchi, Hapuarachchige Chanditha; Harwood, Valerie J.; Rajal, Verónica Beatriz; Jackson, Greg; Khan, Stuart J.; Wuertz, Stefan; Xagoraraki, Irene; Zhang, Qian; Zimmer Faust, Amity G.; Shanks, Orin C.
• Año de publicación: 2022
• Idioma: inglés
• Tipo de recurso: artículo
• Materia: COVID-19; SARS-COV-2; WASTEWATER; SURVEILLANCE
• Institución: Consejo Nacional de Investigaciones Científicas y Técnicas

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) inwastewater can potentially provide an earlywarning 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 inwastewater, culminating in recommended strategies that can be implemented to identify and mitigate some of these errors. Recommendations include stringent QA/QC measures, representative samplingapproaches, 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, particularlywhen the incidence of SARS-CoV-2 inwastewater is low. Corrective and confirmatory actionsmust be in place for inconclusive results or results diverging fromcurrent 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.
Fil: Ahmed, Warish. Commonwealth Scientific And Industrial Research Organisation (csiro);
Fil: Simpson, Stuart L.. Commonwealth Scientific And Industrial Research Organisation (csiro);
Fil: Bertsch, Paul M.. Radboud University Medical Center; Países Bajos
Fil: Bibby, Kyle. University of Notre Dame; Estados Unidos
Fil: Bivins, Aaron. University of Notre Dame; Estados Unidos
Fil: Blackall, Linda L.. University of Melbourne; Australia
Fil: Bofill-Mas, Sílvia. Universidad de Barcelona. Facultad de Biología; España
Fil: Bosch, Albert. Universidad de Barcelona; España
Fil: Brandão, João. Instituto Nacional de Saúde Dr. Ricardo Jorge; Portugal
Fil: Choi, Phil M.. The University of Queensland; Australia
Fil: Ciesielski, Mark. University of North Carolina; Estados Unidos
Fil: Donner, Erica. University Of South Australia; Australia
Fil: D'Souza, Nishita. Michigan State University; Estados Unidos
Fil: Farnleitner, Andreas H.. Technische Universitat Wien; Austria
Fil: Gerrity, Daniel. Southern Nevada Water Authority; Estados Unidos
Fil: Gonzalez, Raul. Hampton Roads Sanitation District; Estados Unidos
Fil: Griffith, John F.. Southern California Coastal Water Research Project; Estados Unidos
Fil: Gyawali, Pradip. Institute Of Environmental Science And Research Ltd; Nueva Zelanda
Fil: Haas, Charles N.. Drexel University; Estados Unidos
Fil: Hamilton, Kerry A.. Arizona State University; Estados Unidos
Fil: Hapuarachchi, Hapuarachchige Chanditha. National Environment Agency; Singapur
Fil: Harwood, Valerie J.. University Of South Florida; Estados Unidos
Fil: Rajal, Verónica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones para la Industria Química. Universidad Nacional de Salta. Facultad de Ingeniería. Instituto de Investigaciones para la Industria Química; Argentina
Fil: Jackson, Greg. Health Protection Branch; Australia
Fil: Khan, Stuart J.. University of New South Wales; Australia
Fil: Wuertz, Stefan. Stellenbosch University; Sudáfrica
Fil: Xagoraraki, Irene. Michigan State University; Estados Unidos
Fil: Zhang, Qian. University of Minnesota; Estados Unidos
Fil: Zimmer Faust, Amity G.. Southern California Coastal Water Research Project; Estados Unidos
Fil: Shanks, Orin C.. United States Environmental Protection Agency; Estados Unidos