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. 2021 Dec 10:799:149386.
doi: 10.1016/j.scitotenv.2021.149386. Epub 2021 Jul 31.

Comparative analysis of rapid concentration methods for the recovery of SARS-CoV-2 and quantification of human enteric viruses and a sewage-associated marker gene in untreated wastewater

Warish Ahmed  1 , Aaron Bivins  2 , Stuart L Simpson  3 , Wendy J M Smith  4 , Suzanne Metcalfe  4 , Brian McMinn  5 , Erin M Symonds  6 , Asja Korajkic  5
Affiliations

Comparative analysis of rapid concentration methods for the recovery of SARS-CoV-2 and quantification of human enteric viruses and a sewage-associated marker gene in untreated wastewater

Warish Ahmed et al. Sci Total Environ. .

Abstract

To support public-health-related disease surveillance and monitoring, it is crucial to concentrate both enveloped and non-enveloped viruses from domestic wastewater. To date, most concentration methods were developed for non-enveloped viruses, and limited studies have directly compared the recovery efficiency of both types of viruses. In this study, the effectiveness of two different concentration methods (Concentrating pipette (CP) method and an adsorption-extraction (AE) method amended with MgCl2) were evaluated for untreated wastewater matrices using three different viruses (SARS-CoV-2 (seeded), human adenovirus 40/41 (HAdV 40/41), and enterovirus (EV)) and a wastewater-associated bacterial marker gene targeting Lachnospiraceae (Lachno3). For SARS-CoV-2, the estimated mean recovery efficiencies were significantly greater by as much as 5.46 times, using the CP method than the AE method amended with MgCl2. SARS-CoV-2 RNA recovery was greater for samples with higher titer seeds regardless of the method, and the estimated mean recovery efficiencies using the CP method were 25.1 ± 11% across ten WWTPs when wastewater samples were seeded with 5 × 104 gene copies (GC) of SARS-CoV-2. Meanwhile, the AE method yielded significantly greater concentrations of indigenous HAdV 40/41 and Lachno3 from wastewater compared to the CP method. Finally, no significant differences in indigenous EV concentrations were identified in comparing the AE and CP methods. These data indicate that the most effective concentration method varies by microbial analyte and that the priorities of the surveillance or monitoring program should be considered when choosing the concentration method.

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

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Conflict of interest statement

Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Unlabelled Image
Graphical abstract
Fig. 1
Fig. 1
Virus concentration methods used in this study.
Fig. 2
Fig. 2
Recovery (mean ± SD) of SARS-CoV-2 RNA from ten wastewater treatment plants (WWTPs) using a concentrating pipette (CP method). Each WWTP was sampled three times.
See this image and copyright information in PMC

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