![]() ![]() To facilitate the adoption and implementation of wastewater surveillance more broadly, it is essential to develop and evaluate flexible workflows that can be readily adopted across a range of settings, community needs, and resource availability (D’Aoust et al. However, small and rural communities are often underrepresented in wastewater surveillance programs, due in part to local resource and infrastructure constraints (D’Aoust et al. A growing number of public health, wastewater utility, commercial, and other laboratories are expanding their capacity to conduct routine wastewater surveillance (Naughton et al. With the success of these initial efforts, wastewater surveillance is now being formally adopted into state-wide and national public health programs. In these ways, wastewater surveillance provides an indiscriminate, noninvasive, and cost-effective approach to community surveillance for both COVID-19 and other diseases.Īs an early response to the COVID-19 pandemic, wastewater surveillance for SARS-CoV-2 was implemented rapidly by research laboratories across the globe (Bivins et al. 2021), and may serve as an early warning system for disease outbreaks or surges in transmission (Xagoraraki and O’Brien 2020, Ahmed et al. ![]() Wastewater surveillance is useful for detection of both asymptomatic and symptomatic cases of infection (Schmitz et al. Previous studies have demonstrated that viral levels in wastewater correlate with COVID-19 incidence and hospitalization rates (Peccia et al. ![]() 2020, Sanjuán and Domingo-Calap 2021, Wu et al. The relationship between SARS-CoV-2 viral levels in wastewater and reported COVID-19 cases has been well-documented (Polo et al. These excreta, collected by the sewage network, can be monitored in downstream wastewaters as a proxy for disease surveillance (Medema et al. Individuals infected with SARS-CoV-2 shed viruses and viral RNA through respiratory fluids, saliva, urine, and stool (Kitajima et al. The coronavirus disease 2019 (COVID-19) is an infectious respiratory disease caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). These results indicate that a direct-extraction-based workflow for SARS-CoV-2 wastewater surveillance can provide informative and actionable results. With this approach, we detected as few as five cases of COVID-19 per 100 000 individuals. To compensate for the method’s high limit of detection (approximately 10 6–10 7 copies l −1 in wastewater), we extracted multiple small-volume replicates of each wastewater sample. N1 and N2 assay positivity, viral concentration, and flow-adjusted daily viral load correlated significantly with per-capita case reports of COVID-19 at the county-level (ρ = 0.69–0.82). SARS-CoV-2 viral RNA was detected in 76% (193/254) of influent samples, and the recovery of the surrogate bovine coronavirus was 42% (IQR: 28%, 59%). Bypassing any concentration step, low volumes (280 µl) of influent wastewater were extracted using a commercial kit, and immediately analyzed by RT-qPCR for the SARS-CoV-2 N1 and N2 gene targets. Composite influent wastewater samples were collected weekly for 1 year between June 2020 and June 2021 in Athens-Clarke County, Georgia, USA. ![]() To address some of these issues, we conducted a longitudinal study implementing a simplified workflow for SARS-CoV-2 detection from wastewater, using a direct column-based extraction approach. Workflows for wastewater surveillance generally rely on concentration steps to increase the probability of detection of low-abundance targets, but preconcentration can substantially increase the time and cost of analyses while also introducing additional loss of target during processing. Wastewater surveillance has proven to be an effective tool to monitor the transmission and emergence of infectious agents at a community scale. ![]()
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