Insights from Wastewater Surveillance of SARS-CoV-2 in Skilled Nursing Facilities: Comparing Virus Concentration Methods for Wastewater and Correlating Wastewater Virus Concentrations with Clinical Infections, Georgia, USA, 2022

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Abstract To understand the utility of healthcare facility-level wastewater surveillance (WWS) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), it is important to correlate wastewater SARS-CoV-2 RNA detection with the number of clinical infections. WWS for SARS-CoV-2 was performed at three skilled nursing facilities (SNFs) over 25 weeks. Electronegative membrane filtration (enMF) and Nanotrap® Magnetic Virus Particles (NP) virus concentration methods were compared. Extracts were tested by droplet digital polymerase chain reaction. Spearman’s correlations (ρ) between wastewater virus RNA concentrations and infection counts were calculated. From split wastewater samples, enMF recovered higher SARS-CoV-2 RNA concentrations than NP. Combining data from all facilities, the median concentrations were 53.0 versus 38.6 gc/100 mL for enMF and NP, respectively (p=0.001). Using enMF, correlations were moderate to strong at SNF A (ρ ranged 0.67 to 0.86, all p-values <0.001). Weak to moderate correlations can be explained by the sampled manhole not representing the entire facility (SNF B, ρ ranged 0.47 to 0.72, p-values ranged <0.001 to 0.12) and longitudinal data gaps from summer heat and equipment maintenance (SNF C, ρ ranged 0.14 to 0.59, p-values ranged 0.52 to <0.01). WWS can be a valuable tool for tracking dynamics of SARS-CoV-2 infections in healthcare facilities. Highlights Healthcare facility-level wastewater surveillance may be useful as a complementary surveillance method for infectious disease targets, including SARS-CoV-2. Two virus concentration methods were compared, electronegative membrane filtration (enMF) and Nanotrap® Magnetic Virus Particles (NP). Spearman’s correlations (ρ) between wastewater virus RNA concentrations and infection counts were calculated among several subpopulations at three skilled nursing facilities. Graphical Abstract

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