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Incorporating SARS-CoV-2 Case Surveillance Data in the Comparison of Wastewater Concentration Methods for Wastewater Surveillance

Published online by Cambridge University Press:  23 June 2026

Florence Whitehill
Affiliation:
CDC
Amanda Lyons
Affiliation:
Centers for Disease Control and Prevention
Ariel J. Santiago
Affiliation:
Centers for Disease Control and Prevention
Christine Ganim
Affiliation:
CDC
Susanna Lenz
Affiliation:
CDC
Paige Gable
Affiliation:
CDC
Magdalena Medrzycki
Affiliation:
CDC
Maroya Walters
Affiliation:
Centers for Disease Control and Prevention
Amelia Keaton
Affiliation:
Centers for Disease Control and Prevention
Angela Coulliette-Salmond
Affiliation:
CDC, U.S. Public Health Service

Abstract

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Background: Healthcare facility-level wastewater surveillance (WWS) is a relatively novel tool for pathogen surveillance. The most sensitive methods for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in wastewater remain unknown. In this study at skilled nursing facilities (SNFs), two wastewater concentration methods were evaluated for SARS-CoV-2 detection by droplet digital polymerase chain reaction (ddPCR): Electronegative membrane filtration (enMF) and Nanotrap® Magnetic Virus Particles (NP). SARS-CoV-2 case surveillance data from the SNFs was incorporated into this evaluation. Methods: Wastewater samples were collected weekly from manholes at three SNFs over 25 weeks in 2022. Samples were split and concentrated by both enMF and NP, then virus concentration was determined by ddPCR. We utilized a weekly questionnaire and each facility’s National Healthcare Safety Network COVID-19 Report to identify the number of resident and healthcare personnel cases in each facility. Differences in the median SARS-CoV-2 concentration were compared across the enMF and NP methods using the Wilcoxon signed-rank test (?=0.05). Spearman’s correlation (?) was used to evaluate the relationship between the SARS-CoV-2 concentration in wastewater and SARS-CoV-2 case counts. Results: For each facility separately, as well as overall, median virus concentration in wastewater from enMF was higher than that from NP (SNF A: 2.84 vs. 1.49 genome copies [gc]/100 mL; SNF B: 8.50 vs. 6.08 gc/100 mL; SNF C: 5.3 vs. 3.86 gc/100 mL; overall: 5.3 vs. 3.86 gc/100 mL). The Wilcoxon signed-rank test confirmed that the difference in median virus concentrations was statistically significant for SNF B (p=0.001) and overall (p=0.001). Spearman’s correlation results showed overall higher correlations with enMF than with NP and a greater number of statistically significant results among subpopulations. For example: using enMF, the correlation among residents and healthcare personnel combined at SNF A was ?=0.86 (p=<0.0001), at SNF B was ?=0.50 (p=<0.01), and at SNF C was ?=0.59 (p=<0.01), whereas using NP, the correlation at SNF A was ?=0.54 (p=<0.01), SNF B was ?=0.46 (p=0.02), and SNF C was ?=0.23 (p=0.26). Conclusions: enMF yielded higher SARS-CoV-2 concentrations than NP and produced stronger correlations between wastewater signal and case counts, though both methods yielded acceptable results. enMF should be considered as an effective virus concentration method for healthcare facility-level WWS to strengthen and complement clinical methods of case detection.

Information

Type
Surveillance
Creative Commons
Creative Common License - CCCreative Common License - BY
This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited.
Copyright
© The Author(s), 2026. Published by Cambridge University Press on behalf of The Society for Healthcare Epidemiology of America