Ahmed, WarishSimpson, StuartBertsch, PaulBibby, KyleBivins, AaronBlackall, LindaBofill-Mas, SilviaBosch, AlbertBrandao, JoaoChoi, PhilCiesielski, MarkDonner, EricaD'Souza, NishitaFarnleitner, AndreasGerrity, DanielGonzalez, RaulGriffith, JohnGyawali, PradipHaas, CharlesHamilton, KerryHapuarachchi, ChandithaHarwood, ValerieHaque, RehnumaJackson, GregKhan, StuartKhan, WesaalKitajima, MasaakiKorajkic, AsjaLa Rosa, GiuseppinaLayton, BlytheLipp, ErinMcLellan, SandraMcMinn, BrianMedema, GertjanMetcalfe, SuzanneMeijer, WimMueller, JochenMurphy, HeatherNaughton, ColleenNoble, RachelPayyappat, SudhiPetterson, SusanPitkanen, TarjaRajal, VeronicaReyneke, BrandonRoman, FernandoRose, JoanRusinol, MartaSadowsky, MichaelSala-Comorera, LauraSetoh, Yin XiangSherchan, SamendraSirikanchana, KwanraweeSmith, WendySteele, JoshuaSabburg, RosalieSymonds, ErinThai, PhongThomas, KevinTynan, JoshToze, SimonThompson, JanelleWhiteley, AndyWong, JudithSano, DaisukeWuertz, StefanXagoraraki, IreneZhang, QianZimmer-Faust, AmityShanks, Orin2022-01-272022-01-272021-08-25Sci Total Environ. 2022 Jan 20;805:149877. doi: 10.1016/j.scitotenv.2021.149877. Epub 2021 Aug 25. Review1879-1026http://hdl.handle.net/10400.18/7884Wastewater surveillance for pathogens using the reverse transcription-polymerase chain reaction (RT-PCR) is an effective, resource-efficient tool for gathering additional community-level public health information, including the incidence and/or prevalence and trends of coronavirus disease-19 (COVID-19). Surveillance of SARS-CoV-2 in wastewater may 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 rapidly increasing. However, there are no standardized protocols nor 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 lead to false-positive and -negative errors in the surveillance of SARS-CoV-2, culminating in recommendations and strategies that can be implemented to identify and mitigate these errors. Recommendations include, stringent QA/QC measures, representative sampling approaches, effective virus concentration and efficient RNA extraction, amplification 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 during a low incidence of SARS-CoV-2 in wastewater. Corrective and confirmatory actions must be in place for inconclusive and/or potentially significant results (e.g., initial onset or reemergence of COVID-19 in a community). It will also be prudent to perform inter-laboratory comparisons to ensure results are reliable and interpretable for ongoing and retrospective analyses. The strategies that are recommended in this review aim to improve SARS-CoV-2 characterization for wastewater surveillance applications. A silver lining of the COVID-19 pandemic is that the efficacy of wastewater surveillance was demonstrated during this global crisis. In the future, wastewater will play an important role in the surveillance of a range of other communicable diseases.Highlights: Harmonized QA/QC procedures for SARS-CoV-2 wastewater surveillance are lacking; Wastewater analysis protocols are not optimized for trace analysis of viruses; False-positive and -negative errors have consequences for public health responses; Inter-laboratory studies utilizing standardized reference materials and protocols are needed.engCOVID-19SARS-CoV-2WastewaterSurveillanceFalse-positiveFalse-negativeRT-PCRjournal article10.1016/j.scitotenv.2021.149877