Targeted Community Wastewater Surveillance for Sars-Cov-2 and Mpox Virus During a Mass-Gathering Event

Wastewater surveillance has emerged recently as a powerful approach to understanding infectious disease dynamics in densely populated zones. Wastewater surveillance, while promising as a public health tool, is often hampered by slow turn-around times, complex analytical protocols, and resource-intensive techniques. This study focused on rapidly quantifying viruses in wastewater during a mass gathering event. Wastewater samples (n = 131) were collected from residential and commercial manholes, pump stations, and a wastewater treatment plant. The presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), mpox (formerly known as monkeypox) virus, and fecal indicator, pepper mild mottle virus (PMMoV) in wastewater was determined using digital PCR. In this study, the use of Nanotrap® Microbiome Particles and microfluidic digital PCR system produced comparable results to other established methodologies, with reduced process complexity and response times, offering an average viral recovery of 10.1% across the three viral gene targets. The targeted wastewater surveillance portion of this study quantified differences in virus concentration between individual sites, types of sample locations, and sample zones. The mpox virus (MPXV) was sporadically detected in wastewater samples during the surveillance period, without distinct temporal trends. SARS-CoV-2 RNA concentrations mirrored the city’s COVID-19 cases, confirming the predictive properties of wastewater surveillance. Wastewater surveillance continues to be beneficial for tracking diseases that display gastrointestinal symptoms, including SARS-CoV-2, and can be a powerful tool for sentinel surveillance. However, careful site selection and a thorough understanding of community dynamics are necessary when performing targeted surveillance during temporary mass-gathering events as potential confirmation bias may occur.