Longitudinal monitoring of SARS-CoV-2 in wastewater using viral genetic markers and the estimation of unconfirmed COVID-19 cases

In this study, wastewater-based surveillance was carried out to establish the correlation between SARS-CoV-2 viral RNA concentrations in wastewater and the incidence of corona virus disease 2019 (COVID-19) from clinical testing. The influent wastewater of three major water reclamation facilities (WRFs) in Northern Nevada, serving a population of 390,750, was monitored for SARS-CoV-2 viral RNA gene markers, N1 and N2, from June 2020 through September 2021. A total of 614 samples were collected and analyzed. The SARS-CoV-2 concentrations in wastewater were observed to peak twice during the study period. A moderate correlation trend between coronavirus disease 2019 (COVID-19) incidence data from clinical testing and SARS-CoV-2 viral RNA concentrations in wastewater was observed (Spearman r = 0.533). This correlation improved when using weekly average SARS-CoV-2 marker concentrations of wastewater and clinical case data (Spearman r = 0.790), presumably by mitigating the inherent variability of the environmental dataset and the effects of clinical testing artifacts (e.g., reporting lags). The research also demonstrated the value of wastewater-based surveillance as an early warning signal for early detection of trends in COVID-19 incidence. This was accomplished by identifying that the reported clinical cases had a stronger correlation to SARS-CoV-2 wastewater monitoring data when they were estimated to lag 7-days behind the wastewater data. The results aided local decision makers in developing strategies to manage COVID-19 in the region and provide a framework for how wastewater-based surveillance can be applied across localities to enhance the public health monitoring of the ongoing pandemic.