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

• Published in: The Science of the total environment Vol. 808; p. 151916
• Main Authors: Kevill, Jessica L., Pellett, Cameron, Farkas, Kata, Brown, Mathew R., Bassano, Irene, Denise, Hubert, McDonald, James E., Malham, Shelagh K., Porter, Jonathan, Warren, Jonathan, Evens, Nicholas P., Paterson, Steve, Singer, Andrew C., Jones, Davey L.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 20.02.2022; The Author(s). Published by Elsevier B.V
• Subjects: ammonium sulfate; COVID-19; data quality; environment; Faecal indicator virus; genome; Humans; monitoring; pandemic; polyethylene glycol; qRT-PCR; RNA detection; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; storage temperature; Surface-Active Agents; surfactants; Temperature; turbidity; ultrafiltration; viruses; Wastewater; Wastewater concentration; COVID-19; Faecal indicator virus; Wastewater concentration; RNA detection; qRT-PCR
• ISSN: 0048-9697; 1879-1026; 1879-1026
• DOI: 10.1016/j.scitotenv.2021.151916

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. [Display omitted] •Comparison of wastewater viral recovery using three concentration methods.•Control experiment looked at effect of turbidity and surfactant on viral recoveries.•No sig. difference between wastewater viral recovery methods.•Sequencing result comparable between viral concentration methods.•Solids and surfactant impact viral recovery dependant on concentration method.