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 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.
Keywords: COVID-19; False negative; False positive; RT-PCR; SARS-CoV-2; Surveillance; Wastewater.
【저자키워드】 COVID-19, SARS-CoV-2, RT-PCR, wastewater, Surveillance, False positive, False negative, 【초록키워드】 coronavirus disease, public health, Efficacy, reliability, COVID-19 pandemic, severe acute respiratory syndrome coronavirus-2, severe acute respiratory syndrome Coronavirus, RT-PCR, virus, Laboratory, RNA extraction, Severe acute respiratory syndrome, polymerase chain reaction, wastewater, PCR, Measures, Coronavirus disease-19, pathogen, COVID-19 infection, comparison, communicable diseases, Community, Pathogens, reverse transcription, SARS-CoV-2 RNA, health information, interpretability, incidence, Quality control, respiratory, silver, information, False positive, Critical, false-negative, RT-PCR assay, False-positive, False negative, Quality assurance, Reverse transcription-polymerase chain reaction, retrospective, Concentration, Chain Reaction, Factor, approaches, COVID-19 infections, standardized protocols, false, positive, standardized protocol, data interpretation, Inclusion, Inconclusive, mitigate, controls, effective, initial, IMPROVE, identify, include, demonstrated, analyses, 【제목키워드】 quantification, RT-PCR detection,