Wastewater-Based Epidemiology of SARS-CoV-2 RNA in Bethlehem, PA and Lehigh University

Wastewater surveillance for SARS-CoV-2 provides a broad assessment of community health since wastewater represents all community members, regardless of health care access and status of health (i.e., symptomatic and asymptomatic). Wastewater surveillance also provides early detection of disease transmission since the virus can be present in human waste before the presentation of clinical symptoms. We analyzed wastewater from Lehigh University (Bethlehem, PA) as well as the greater Bethlehem community for SARS-CoV-2 (N1, N2, and E genes) from August 2020 through May 2021. Total RNA was extracted and analyzed for SARS-CoV-2 and pepper mild mottle virus (PMMoV) by RT-qPCR. Of 73 Bethlehem wastewater samples, the number of positive samples depended on which SARS-CoV-2 gene was screened: 62 (84.9%) for N1, 52 (71.2%) for E, and 28 (38.4%) for N2. Of 67 university wastewater samples, the numbers of positive samples were 59 (88.1%) for N1, 51 (76.1%) for E, and 31 (46.3%) for N2. Temporal trends of SARS-CoV-2 in wastewater mirrored trends in COVID-19 positive cases in the Bethlehem community. Normalizing SARS-CoV-2 RNA concentrations to PMMoV (a human fecal indicator) increased the correlation between both N1 (ρ increased from 0.37 to 0.72) and E (ρ increased from 0.32 to 0.61) concentrations in wastewater with COVID-19 cases in the community.