Comparison of virus concentration methods for the RT-qPCR-based recovery of murine hepatitis virus, a surrogate for SARS-CoV-2 from untreated wastewater

Warish Ahmed; Paul M Bertsch; Aaron Bivins; Kyle Bibby; Kata Farkas; Amy Gathercole; Eiji Haramoto; Pradip Gyawali; Asja Korajkic; Brian R McMinn; Jochen F Mueller; Stuart L Simpson; Wendy J M Smith; Erin M Symonds; Kevin V Thomas; Rory Verhagen; Masaaki Kitajima

doi:10.1016/j.scitotenv.2020.139960

Comparison of virus concentration methods for the RT-qPCR-based recovery of murine hepatitis virus, a surrogate for SARS-CoV-2 from untreated wastewater

Sci Total Environ. 2020 Oct 15:739:139960. doi: 10.1016/j.scitotenv.2020.139960. Epub 2020 Jun 5.

Authors

Warish Ahmed¹, Paul M Bertsch², Aaron Bivins³, Kyle Bibby³, Kata Farkas⁴, Amy Gathercole⁵, Eiji Haramoto⁶, Pradip Gyawali⁷, Asja Korajkic⁸, Brian R McMinn⁸, Jochen F Mueller⁹, Stuart L Simpson¹⁰, Wendy J M Smith¹¹, Erin M Symonds¹², Kevin V Thomas⁹, Rory Verhagen⁹, Masaaki Kitajima¹³

Affiliations

¹ CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Dutton Park, QLD, 4102, Australia. Electronic address: Warish.Ahmed@csiro.au.
² CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Dutton Park, QLD, 4102, Australia.
³ Environmental Change Initiative, University of Notre Dame, 721 Flanner Hall, Notre Dame, IN 46556, USA.
⁴ School of Ocean Sciences, Bangor University, Menai Bridge, Anglesey LL59 5AB, UK.
⁵ ComPath, South Australian Health and Medical Research Institute, Adelaide, SA 5000, Australia.
⁶ Interdisciplinary Center for River Basin Environment, University of Yamanashi, 4 - 3 -11 Takeda, Kofu, Yamanashi 400-8511, Japan.
⁷ Institute of Environmental Science and Research Ltd. (ESR), Porirua 5240, New Zealand.
⁸ United States Environmental Protection Agency, Office of Research and Development, 26W Martin Luther King Jr. Drive, Cincinnati, OH 45268, USA.
⁹ Queensland Alliance for Environmental Health Sciences (QAEHS), University of Queensland, 20 Cornwall Street, Woolloongabba, QLD 4102, Australia.
¹⁰ CSIRO Land and Water, Lucas Heights, NSW 2234, Australia.
¹¹ CSIRO Agriculture and Food, Queensland Bioscience Precinct, St Lucia, QLD 4067, Australia.
¹² College of Marine Science, University of South Florida, 140 Seventh Avenue South, St. Petersburg, FL 33701, USA.
¹³ Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, North 13 West 8, Kita-ku, Sapporo, Hokkaido 060-0032, Japan.

Abstract

There is currently a clear benefit for many countries to utilize wastewater-based epidemiology (WBE) as part of ongoing measures to manage the coronavirus disease 2019 (COVID-19) global pandemic. Since most wastewater virus concentration methods were developed and validated for nonenveloped viruses, it is imperative to determine the efficiency of the most commonly used methods for the enveloped severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Municipal wastewater seeded with a human coronavirus (CoV) surrogate, murine hepatitis virus (MHV), was used to test the efficiency of seven wastewater virus concentration methods: (A-C) adsorption-extraction with three different pre-treatment options, (D-E) centrifugal filter device methods with two different devices, (F) polyethylene glycol (PEG 8000) precipitation, and (G) ultracentrifugation. MHV was quantified by reverse-transcription quantitative polymerase chain reaction and the recovery efficiency was calculated for each method. The mean MHV recoveries ranged from 26.7 to 65.7%. The most efficient methods were adsorption-extraction methods with MgCl₂ pre-treatment (Method C), and without pre-treatment (Method B). The third most efficient method used the Amicon® Ultra-15 centrifugal filter device (Method D) and its recovery efficiency was not statistically different from the most efficient methods. The methods with the worst recovery efficiency included the adsorption-extraction method with acidification (A), followed by PEG precipitation (F). Our results suggest that absorption-extraction methods with minimal or without pre-treatment can provide suitably rapid, cost-effective and relatively straightforward recovery of enveloped viruses in wastewater. The MHV is a promising process control for SARS-CoV-2 surveillance and can be used as a quality control measure to support community-level epidemic mitigation and risk assessment.

Keywords: COVID-19; Concentration method; Enveloped virus; Filtration; Murine hepatitis virus; Recovery; SARS-CoV-2; Untreated wastewater.

MeSH terms

Animals
Betacoronavirus
COVID-19
Coronavirus Infections*
Humans
Mice
Murine hepatitis virus*
Pandemics*
Pneumonia, Viral*
SARS-CoV-2
Viruses*
Wastewater

Substances

Waste Water