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SARS-CoV-2 in wastewater settled solids is associated with COVID-19 cases in a large urban sewershed

By Katherine Graham, Stephanie Loeb, Marlene Wolfe, David Catoe, Nasa Sinnott-Armstrong, Sooyeol Kim, Kevan Yamahara, Lauren Sassoubre, Lorelay Mendoza, Laura Roldan-Hernandez, Linlin Li, Krista Wigginton, Alexandria Boehm

Posted 15 Sep 2020
medRxiv DOI: 10.1101/2020.09.14.20194472

Wastewater-based epidemiology (WBE) may be useful for informing public health response to viral diseases like COVID-19 caused by SARS-CoV-2. We quantified SARS-CoV-2 RNA in wastewater influent and primary settled solids in two wastewater treatment plants to inform the pre-analytical and analytical approaches, and to assess whether influent or solids harbored more viral targets. The primary settled solids samples resulted in higher SARS-CoV-2 detection frequencies than the corresponding influent samples. Likewise, SARS-CoV-2 RNA was more readily detected in solids using one-step digital droplet (dd)RT-PCR than with two-step RT-QPCR and two-step ddRT-PCR, likely owing to reduced inhibition with the one-step ddRT-PCR assay. We subsequently analyzed a longitudinal time series of 89 settled solids samples from a single plant for SARS-CoV-2 RNA as well as coronavirus recovery (bovine coronavirus) and fecal strength (pepper mild mottle virus, PMMoV) controls. SARS-CoV-2 RNA targets N1 and N2 concentrations correlate positively and significantly with COVID-19 clinical confirmed case counts in the sewershed. Together, the results demonstrate that measuring SARS-CoV-2 RNA concentrations in settled solids may be a more sensitive approach than measuring SARs-CoV-2 in influent.

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