Evidence map›Paper›PMID 34818780›Full record

ReviewThe Science of the total environment2022

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

Warish Ahmed, Stuart L Simpson, Paul M Bertsch, Kyle Bibby, Aaron Bivins, Linda L Blackall, Sílvia Bofill-Mas, Albert Bosch, João Brandão, Phil M Choi and 60 more

Open access · hybridAbstract readReview
PubMed Full text at PMC Publisher
In one paragraph

Review in The Science of the total environment, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 129 papers, 4 of them syntheses that pooled it.

0numbers the graph read from it
0cells of the map it votes in
129citing papers in PubMed, 4 pooled it
14.8field-weighted citation impact, top 1% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

129 citing papers in PubMed, 4 syntheses or guidelines pooled it, 274 citations in OpenAlex.

  1. Pooled it
  2. Pooled it
  3. Pooled it
  4. Detecting SARS-CoV-2 in sludge samples: A systematic review.The Science of the total environment · 2023
    Pooled it
  5. Review
  6. Observational
  7. Article
  8. Article
  9. Plastics as vectors for pathogens and antibiotic resistance genes in aquatic systems.Water science and technology : a journal of the International Association on Water Pollution Research · 2026
    Review
  10. Article
  11. Article
  12. Review
  13. Article
  14. Review
  15. Absence of hepatitis E virus RNA in bovines from Portugal.Veterinary research communications · 2025
    Article
  16. Article
  17. Review
  18. Review
  19. Article
  20. Article

69 more citing papers are in PubMed but not listed here.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

70 authors at 20 institutions in 16 countries.

Warish AhmedCSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia. Electronic address: Warish.Ahmed@csiro.au.
Stuart L SimpsonCSIRO Land and Water, Lucas Heights, NSW 2234, Australia.
Paul M BertschCSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia.
Kyle BibbyDepartment of Civil & Environmental Engineering & Earth Science, University of Notre Dame, 156 Fitzpatrick Hall, Notre Dame, IN 46556, USA.
Aaron BivinsDepartment of Civil & Environmental Engineering & Earth Science, University of Notre Dame, 156 Fitzpatrick Hall, Notre Dame, IN 46556, USA.
Linda L BlackallSchool of BioSciences, The University of Melbourne, Melbourne, VIC, Australia.
Sílvia Bofill-MasLaboratory of Virus Contaminants of Water and Food, Department of Genetics, Microbiology and Statistics, Faculty of Biology, University of Barcelona, 08028 Barcelona, Spain.
Albert BoschEnteric Virus Laboratory, Department of Genetics, Microbiology and Statistics, University of Barcelona, Avda. Diagonal 643, 08028 Barcelona, Spain.
João BrandãoDepartment of Environmental Health, National Institute of Health Dr. Ricardo Jorge, Lisboa, Portugal.
Phil M ChoiWater Unit, Health Protection Branch, Prevention Division, Queensland Health, QLD, Australia; The University of Queensland, Queensland Alliance for Environmental Health Sciences, QLD, Australia.
Mark CiesielskiUniversity of North Carolina at Chapel Hill, Institute of Marine Sciences, Morehead City, NC, United States.
Erica DonnerFuture Industries Institute, University of South Australia, University Boulevard, Mawson Lakes, SA 5095, Australia.
Nishita D'SouzaDepartment of Fisheries and Wildlife, Michigan State University, E. Lansing, MI, USA.
Andreas H FarnleitnerInstitute of Chemical, Environmental & Bioscience Engineering, Research Group Environmental Microbiology and Molecular Diagnostic, 166/5/3, Technische Universität Wien, Vienna, Austria; Research Division Water Quality and Health, Department Pharmacology, Physiology and Microbiology, Karl Landsteiner University of Health Sciences, Dr. Karl-Dorrek-Straβe 30, 3500 Krems an der Donau, Austria.
Daniel GerritySouthern Nevada Water Authority, P.O. Box 99954, Las Vegas, NV 89193, USA.
Raul GonzalezHampton Roads Sanitation District, 1434 Air Rail Avenue, Virginia Beach, VA 23455, USA.
John F GriffithSouthern California Coastal Water Research Project, Costa Mesa, CA 92626, USA.
Pradip GyawaliInstitute of Environmental Science and Research Ltd (ESR), Porirua 5240, New Zealand.
Charles N HaasDrexel University, Philadelphia, PA, USA.
Kerry A HamiltonSchool of Sustainable Engineering and the Built Environment and The Biodesign Institute Center for Environmental Health Engineering, Arizona State University, Tempe, AZ 85287, USA.
Hapuarachchige Chanditha HapuarachchiEnvironmental Health Institute, National Environment Agency, Singapore.
Valerie J HarwoodDepartment of Integrative Biology, University of South Florida, Tampa, FL, USA.
Rehnuma HaqueEnvironmental Interventions Unit, Icddr,b, 68 Shaheed Tajuddin Ahmed Sarani, Mohakhali, Dhaka 1212, Bangladesh.
Greg JacksonWater Unit, Health Protection Branch, Prevention Division, Queensland Health, QLD, Australia.
Stuart J KhanWater Research Centre, School of Civil and Environmental Engineering, University of New South Wales, NSW 2052, Australia.
Wesaal KhanDepartment of Microbiology, Faculty of Science, Stellenbosch University, Private Bag X1, Stellenbosch 7602, South Africa.
Masaaki KitajimaDivision of Environmental Engineering, Faculty of Engineering, Hokkaido University, North 13 West 8, Kita-ku, Sapporo, Hokkaido 060-8628, Japan.
Asja KorajkicUnited States Environmental Protection Agency, Office of Research and Development, 26W Martin Luther King Jr. Drive, Cincinnati, OH 45268, USA.
Giuseppina La RosaDepartment of Environment and Health, Istituto Superiore di Sanità, Rome, Italy.
Blythe A LaytonDepartment of Research & Innovation, Clean Water Services, Hillsboro, OR, USA.
Erin LippEnvironmental Health Sciences Department, University of Georgia, Athens, GA 30602, USA.
Sandra L McLellanSchool of Freshwater Sciences, University of Wisconsin-Milwaukee, WI, USA.
Brian McMinnUnited States Environmental Protection Agency, Office of Research and Development, 26W Martin Luther King Jr. Drive, Cincinnati, OH 45268, USA.
Gertjan MedemaKWR Water Research Institute, Groningenhaven 7, 3433 PE Nieuwegein, the Netherlands.
Suzanne MetcalfeCSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia.
Wim G MeijerUCD School of Biomolecular and Biomedical Science, University College Dublin, Dublin, Ireland.
Jochen F MuellerThe University of Queensland, Queensland Alliance for Environmental Health Sciences, QLD, Australia.
Heather MurphyDepartment of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada.
Coleen C NaughtonUniversity of California Merced, Department of Civil and Environmental Engineering, 5200 N. Lake Rd., Merced, CA 95343, USA.
Rachel T NobleUniversity of North Carolina at Chapel Hill, Institute of Marine Sciences, Morehead City, NC, United States.
Sudhi PayyappatSydney Water, 1 Smith Street, Parramatta, NSW 2150, Australia.
Susan PettersonWater and Health Pty Ltd., 13 Lord St, North Sydney, NSW 2060, Australia; School of Medicine, Griffith University, Parklands Drive, Gold Coast, Australia.
Tarja PitkänenFinnish Institute for Health and Welfare, Expert Microbiology Unit, P.O. Box 95, FI-70701 Kuopio, Finland; University of Helsinki, Faculty of Veterinary Medicine, Department of Food Hygiene and Environmental Health, P.O. Box 66, FI-00014, Finland.
Veronica B RajalFacultad de Ingeniería and Instituto de Investigaciones para la Industria Química (INIQUI) - CONICET and Universidad Nacional de Salta, Av. Bolivia 5150, Salta, Argentina.
Brandon ReynekeDepartment of Microbiology, Faculty of Science, Stellenbosch University, Private Bag X1, Stellenbosch 7602, South Africa.
Fernando A RomanUniversity of California Merced, Department of Civil and Environmental Engineering, 5200 N. Lake Rd., Merced, CA 95343, USA.
Joan B RoseDepartment of Fisheries and Wildlife, Michigan State University, E. Lansing, MI, USA.
Marta RusiñolInstitute of Environmental Assessment & Water Research (IDAEA), CSIC, Barcelona, Spain.
Michael J SadowskyBiotechnology Institute and Department of Soil, Water, and Climate, University of Minnesota, St. Paul, MN, USA.
Laura Sala-ComoreraUCD School of Biomolecular and Biomedical Science, University College Dublin, Dublin, Ireland.
Yin Xiang SetohEnvironmental Health Institute, National Environment Agency, Singapore.
Samendra P SherchanDepartment of Environmental Health Sciences, Tulane University, 1440 Canal Street, New Orleans, LA 70112, USA.
Kwanrawee SirikanchanaResearch Laboratory of Biotechnology, Chulabhorn Research Institute, 54 Kampangpetch 6 Road, Laksi, Bangkok 10210, Thailand.
Wendy SmithCSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia.
Joshua A SteeleSouthern California Coastal Water Research Project, Costa Mesa, CA 92626, USA.
Rosalie SabburgCSIRO Agriculture and Food, Bioscience Precinct, St Lucia, QLD 4067, Australia.
Erin M SymondsCollege of Marine Science, University of South Florida, St. Petersburg, FL, USA.
Phong ThaiThe University of Queensland, Queensland Alliance for Environmental Health Sciences, QLD, Australia.
Kevin V ThomasThe University of Queensland, Queensland Alliance for Environmental Health Sciences, QLD, Australia.
Josh TynanThe University of Queensland, Queensland Alliance for Environmental Health Sciences, QLD, Australia.
Simon TozeCSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia.
Janelle ThompsonAsian School of the Environment, Nanyang Technological University, Singapore 639798, Singapore; Singapore Centre for Environmental Life Sciences Engineering (SCELSE) Singapore 637551.
Andy S WhiteleyCSIRO Land and Water, Floreat, WA 6012, Australia.
Judith Chui Ching WongEnvironmental Health Institute, National Environment Agency, Singapore.
Daisuke SanoDepartment of Civil and Environmental Engineering, Graduate School of Engineering, Tohoku University, Aoba 6-6-06, Aramaki, Aoba-Ku, Sendai, Miyagi 980-8597, Japan.
Stefan WuertzSingapore Centre for Environmental Life Sciences Engineering (SCELSE) Singapore 637551; School of Civil and Environmental Engineering, Nanyang Technological University, Singapore 639798.
Irene XagorarakiDepartment of Civil and Environmental Engineering, Michigan State University, East Lansing, MI 48824, USA.
Qian ZhangBiotechnology Institute and Department of Soil, Water, and Climate, University of Minnesota, St. Paul, MN, USA.
Amity G Zimmer-FaustSouthern California Coastal Water Research Project, Costa Mesa, CA 92626, USA.
Orin C ShanksUnited States Environmental Protection Agency, Office of Research and Development, 26W Martin Luther King Jr. Drive, Cincinnati, OH 45268, USA.
University of Helsinki · FICSIRO Land and Water · AUEnvironmental Protection Agency · USMichigan State University · USTulane University · USAgriculture and Food · AUCollege of Marin · USSouthern California Coastal Water Research Project · USSydney Water · AUTohoku University · JPUniversitat de Barcelona · ESUniversity of California, Merced · USUniversity of North Carolina at Chapel Hill · USUniversity of Notre Dame · USUniversity of Wisconsin–Milwaukee · USBiotechnology Institute · USCenter for Environmental Health · USChulabhorn Research Institute · THDrexel University · USHampton Roads Sanitation District · US

Funding

Research Mentoring and Building Capacity of underrepresented Minority Research Scientists in India.D43TW010540 · FIC · YALE UNIVERSITY · PI MICHELE BARRY, Eva Harris · 2017 to 2026
$15.5M
FIC NIH HHS D43 TW010540
6 · The paper itself

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.

Indexed as

COVID-19PandemicsHumansProspective StudiesReproducibility of ResultsRetrospective StudiesReverse Transcriptase Polymerase Chain ReactionRNA, ViralSARS-CoV-2WastewaterWastewater-Based Epidemiological MonitoringRNA, ViralWastewaterCOVID-19False negativeFalse positiveRT-PCRSARS-CoV-2SurveillanceWastewater

Identifiers

PMID34818780
PMCPMC8386095
OpenAlexW3153383196

What OpenQuestion holds

Textmetadata
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.