Evaluating various composite sampling modes for detecting pathogenic SARS-CoV-2 virus in raw sewage

Li, Ye; Ash, Kurt T.; Joyner, Dominique C.; Williams, Daniel E.; Alamilla, Isabella; McKay, Peter J.; Iler, Chris; Hazen, Terry C.

doi:10.3389/fmicb.2023.1305967

Title: Evaluating various composite sampling modes for detecting pathogenic SARS-CoV-2 virus in raw sewage

Journal Article · 22 November 2023 · Frontiers in Microbiology

DOI: https://doi.org/10.3389/fmicb.2023.1305967 · OSTI ID:2283822

Li, Ye ^[1];

^[2]; Joyner, Dominique C. ^[2]; Williams, Daniel E. ^[2]; Alamilla, Isabella ^[1]; McKay, Peter J. ^[1]; Iler, Chris ^[1];

^[2]

Univ. of Tennessee, Knoxville, TN (United States)
Univ. of Tennessee, Knoxville, TN (United States); Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Inadequate sampling approaches to wastewater analyses can introduce biases, leading to inaccurate results such as false negatives and significant over- or underestimation of average daily viral concentrations, due to the sporadic nature of viral input. To address this challenge, we conducted a field trial within the University of Tennessee residence halls, employing different composite sampling modes that encompassed different time intervals (1 h, 2 h, 4 h, 6 h, and 24 h) across various time windows (morning, afternoon, evening, and late-night). Our primary objective was to identify the optimal approach for generating representative composite samples of SARS-CoV-2 from raw wastewater. Utilizing reverse transcription-quantitative polymerase chain reaction, we quantified the levels of SARS-CoV-2 RNA and pepper mild mottle virus (PMMoV) RNA in raw sewage. Our findings consistently demonstrated that PMMoV RNA, an indicator virus of human fecal contamination in water environment, exhibited higher abundance and lower variability compared to pathogenic SARS-CoV-2 RNA. Significantly, both SARS-CoV-2 and PMMoV RNA exhibited greater variability in 1 h individual composite samples throughout the entire sampling period, contrasting with the stability observed in other time-based composite samples. Through a comprehensive analysis of various composite sampling modes using the Quade Nonparametric ANCOVA test with date, PMMoV concentration and site as covariates, we concluded that employing a composite sampler during a focused 6 h morning window for pathogenic SARS-CoV-2 RNA is a pragmatic and cost-effective strategy for achieving representative composite samples within a single day in wastewater-based epidemiology applications. This method has the potential to significantly enhance the accuracy and reliability of data collected at the community level, thereby contributing to more informed public health decision-making during a pandemic.

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Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 2283822

Journal Information:: Frontiers in Microbiology, Journal Name: Frontiers in Microbiology Journal Issue: 2023 Vol. 14; ISSN 1664-302X

Publisher:: Frontiers Research FoundationCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
SARS-CoV-2 RNA
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raw sewage
sampling modes
sampling duration
sampling timing

Title: Evaluating various composite sampling modes for detecting pathogenic SARS-CoV-2 virus in raw sewage

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References (58)

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