SARS-CoV-2 RNA and N Antigen Quantification via Wastewater at the Campus Level, Building Cluster Level, and Individual-Building Level

• Type of Material: Article
• Authors: Winnie Zambrana, Stanford University; David Catoe, SLAC National Accelerator Laboratory; Mhara M Coffman, Stanford University; Sooyeol Kim, Stanford University; Archana Anand, Stanford University; Daniel Solis, Stanford University; Malaya K Sahoo, Stanford University; Benjamin A Pinsky, Stanford University; Ami S Bhatt, Stanford University; Alexandria B Boehm, Stanford University; Marlene Wolfe, Emory University
• Language: English
• Date: 2022-01-01
• Publisher: ACS Publications
• Publication Version: Final Published Version
• Copyright Statement: © 2022 American Chemical Society
• License: Creative Commons Attribution-NonCommercial 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1021/acsestwater.2c00050
• Title of Journal or Parent Work: ACS Environmental Science and Technology Water
• Grant/Funding Information: This work was supported, in part, by Meso Scale Diagnostics (MSD), who provided the S-PLEX SARS-CoV-2 N Kits, BioTek 405 Select automated 96-well plate washer, and MESO QuickPlex SQ 120 Reader used in this study. MSD had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.; This work was funded by the Provost’s Office at Stanford University.
• Keywords: building level monitoring; SARS-CoV-2; campus monitoring; antigen assay; wastewater surveillance
• Research Categories: Health Sciences, Medicine and Surgery; Health Sciences, Pathology

Abstract

Monitoring wastewater for SARS-CoV-2 from populations smaller than those served by wastewater treatment plants may help identify small spatial areas (subsewersheds) where COVID-19 infections are present. We sampled wastewater from three nested locations with different sized populations within the same sewer network at a university campus and quantified SARS-CoV-2 RNA using reverse transcriptase droplet digital polymerase chain reaction (PCR). SARS-CoV-2 RNA concentrations and/or concentrations normalized by PMMoV were positively associated with laboratory-confirmed COVID-19 cases for both the sewershed level and the subsewershed level. We also used an antigen-based assay to detect the nucleocapsid (N) antigen from SARS-CoV-2 in wastewater samples at the sewershed level. The N antigen was regularly detected at the sewershed level, but the results were not associated with either laboratory-confirmed COVID-19 cases or SARS-CoV-2 RNA concentrations. The results of this study indicate that wastewater monitoring based on quantification of SARS-CoV-2 RNA using PCR-based methods is associated with COVID-19 cases at multiple geographic scales within the subsewershed level and can serve to aid the public health response.