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. 2023 Feb 10;859(Pt 1):160072.
doi: 10.1016/j.scitotenv.2022.160072. Epub 2022 Nov 8.

Comparative analysis of Adsorption-Extraction (AE) and Nanotrap® Magnetic Virus Particles (NMVP) workflows for the recovery of endogenous enveloped and non-enveloped viruses in wastewater

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

Comparative analysis of Adsorption-Extraction (AE) and Nanotrap® Magnetic Virus Particles (NMVP) workflows for the recovery of endogenous enveloped and non-enveloped viruses in wastewater

Warish Ahmed et al. Sci Total Environ. .

Abstract

In this study, two virus concentration methods, namely Adsorption-Extraction (AE) and Nanotrap® Magnetic Virus Particles (NMVP) along with commercially available extraction kits were used to quantify endogenous pepper mild mottle virus (PMMoV) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in nucleic acid extracted from 48 wastewater samples collected over six events from eight wastewater treatment plants (WWTPs). The main aim was to determine which workflow (i.e., concentration and extraction methods) produces greater concentrations of endogenous PMMoV and SARS-CoV-2 gene copies (GC) in comparison with each other. Turbidity and total suspended solids (TSS) of wastewater samples within and among the eight WWTPs were highly variable (41-385 NTU and 77-668 mg/L TSS). In 58 % of individual wastewater samples, the log10 GC concentrations of PMMoV were greater by NMVP workflow compared to AE workflow. Paired measurements of PMMoV GC/10 mL from AE and NMVP across all 48 wastewater samples were weakly correlated (r = 0.455, p = 0.001) and demonstrated a poor linear relationship (r2 = 0.207). The log10 GC concentrations of SARS-CoV-2 in 69 % of individual samples were greater by AE workflow compared to NMVP workflow. In contrast to PMMoV, the AE and NMVP derived SARS-CoV-2 GC counts were strongly correlated (r = 0.859, p < 0.001) and demonstrated a strong linear relationship (r2 = 0.738). In general, the PMMoV GC achieved by the NMVP workflow decreased with increasing turbidity, but the PMMoV GC by the AE workflow did not appear to be as sensitive to either turbidity or TSS levels. These findings suggest that wastewater sample turbidity or suspended solids concentration, and the intended target for analysis should be considered when validating an optimal workflow for wastewater surveillance of viruses.

Keywords: Concentration methods; Health risks; PMMoV; 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

Fig. 1.
Fig. 1.
Linear relationship between turbidity (NTU) and total suspended solids (TSS) (mg/L) as observed in 48 untreated wastewater samples from eight WWTPs (8 samples per WWTP).
Fig. 2.
Fig. 2.
Observed Cq values from RT-qPCR of the inhibition control (MHV) seeded into purified nucleic acids derived from six untreated wastewater samples per WWTP shown.
Fig. 3.
Fig. 3.
Comparative log10 GC/10 mL of pepper mild mottle virus (PMMoV) in 48 wastewater samples from eight WWTPs using Adsorption Extraction (AE) and Nanotrap® Magnetic Virus Particles (NMVP) methods.
Fig. 4.
Fig. 4.
Comparative log10GC/10 mL ofSARS-CoV-2 in 48 wastewater samples from eight WWTPs using Adsorption Extraction (AE) and Nanotrap®Magnetic Virus Particles (NMVP) methods.
Fig. 5.
Fig. 5.
Correlation (log10 GC/10 mL) between Adsorption Extraction (AE) and Nanotrap® Magnetic Virus Particles (NMVP) as observed in 48 untreated wastewater samples from eight WWTPs (8 samples per WWTP).
Fig. 6.
Fig. 6.
Trends between viruses (PMMoV and SARS-CoV-2) and wastewater turbidity (NTU) and total suspended solids (TSS).
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