A comparison of precipitation and filtration-based SARS-CoV-2 recovery methods and the influence of temperature, turbidity, and surfactant load in urban wastewater

Jessica L Kevill, Cameron Pellett, Kata Farkas, Mathew R Brown, Irene Bassano, Hubert Denise, James E McDonald, Shelagh K Malham, Jonathan Porter, Jonathan Warren, Nicholas P Evens, Steve Paterson, Andrew C Singer, Davey L Jones

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Wastewater-based epidemiology (WBE) has become a complimentary surveillance tool during the SARS-CoV-2 pandemic. Viral concentration methods from wastewater are still being optimised and compared, whilst viral recovery under different wastewater characteristics and storage temperatures remains poorly understood. Using urban wastewater samples, we tested three viral concentration methods; polyethylene glycol precipitation (PEG), ammonium sulphate precipitation (AS), and CP select™ InnovaPrep® (IP) ultrafiltration. We found no major difference in SARS-CoV-2 and faecal indicator virus (crAssphage) recovery from wastewater samples (n = 46) using these methods, PEG slightly (albeit non-significantly), outperformed AS and IP for SARS-CoV-2 detection, as a higher genome copies per litre (gc/l) was recorded for a larger proportion of samples. Next generation sequencing of 8 paired samples revealed non-significant differences in the quality of data between AS and IP, though IP data quality was slightly better and less variable. A controlled experiment assessed the impact of wastewater suspended solids (turbidity; 0–400 NTU), surfactant load (0–200 mg/l), and storage temperature (5–20 °C) on viral recovery using the AS and IP methods. SARS-CoV-2 recoveries were >20% with AS and <10% with IP in turbid samples, whilst viral recoveries for samples with additional surfactant were between 0–18% for AS and 0–5% for IP. Turbidity and sample storage temperature combined had no significant effect on SARS-CoV-2 recovery (p > 0.05), whilst surfactant and storage temperature combined were significant negative correlates (p < 0.001 and p < 0.05, respectively). In conclusion, our results show that choice of methodology had small effect on viral recovery of SARS-CoV-2 and crAssphage in wastewater samples within this study. In contrast, sample turbidity, storage temperature, and surfactant load did affect viral recovery, highlighting the need for careful consideration of the viral concentration methodology used when working with wastewater samples.

Original languageEnglish
Article number151916
JournalScience of the Total Environment
Early online date24 Nov 2021
Publication statusPublished - 20 Feb 2022

Bibliographical note

Funding Information:
Funding was provided by the UK Natural Environment Research Council National under the COVID-19 Wastewater Epidemiology Surveillance Programme ( NE/V010441/1 ) and the Centre for Environmental Biotechnology Project funded though the European Regional Development Fund (ERDF) by Welsh Government. We particularly thank Tony Harrington at Dŵr Cymru-Welsh Water and Jo Harrison and Charmian Abbott at United Utilities PLC alongside staff at the wastewater treatment facilities for their support in this project. We thank Prof Ian Goodfellow (University of Cambridge, UK) for providing MNV and BV2 stocks.

Publisher Copyright:
© 2021 The Authors


  • COVID-19
  • Faecal indicator virus
  • RNA detection
  • Wastewater concentration
  • qRT-PCR
  • Surface-Active Agents
  • SARS-CoV-2
  • Temperature
  • Humans
  • Waste Water

ASJC Scopus subject areas

  • Pollution
  • Waste Management and Disposal
  • Environmental Engineering
  • Environmental Chemistry


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