Emulsion-free Digital PCR Measurement of Wastewater Related Targets using the SARS-CoV-2 Wastewater Surveillance 4-plex Assay

Background

Wastewater based epidemiology (WBE) enables tracking of biomarkers for specific pathogens to monitor for disease outbreak and spread. WBE’s utility in disease surveillance has been proven to be effective in monitoring for rare cases of disease.(1) To effectively monitor and quantity of SARS-CoV-2 viral targets from wastewater samples, it is critical to maximize the amount of information per testing reaction, minimize reagent waste, and control for external factors such as population size and sample processing efficiency.

Measuring wastewater related targets alongside robust controls can provide tangible metrics for normalization of results and help limit the impact of sample preparation variability. The SARS-CoV-2 Wastewater Surveillance 4-plex Assay was designed to detect the N1 and N2 SARS-CoV-2 viral RNA targets alongside the human fecal control, pepper mild mottle virus (PMMoV), and a spike-in sample process control, bovine coronavirus (BCoV).

In this study, we demonstrate SARS-CoV-2 detection and quantification alongside the normalization controls for four wastewater samples collected by the University of Arizona WEST center during their wastewater epidemiology testing efforts.

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  1. Asghar H, et al., Environmental surveillance for polioviruses in the global polio eradication initiative. J Infect Dis. 2014;210:S294–S303.

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