SARS-CoV-2 Protein in Wastewater Mirrors COVID-19 Prevalence

Abstract

The COVID-19 pandemic has given rise to diverse approaches to track infections. The causative agent, SARS-CoV-2 is a fecally-shed RNA virus, and many groups have assayed wastewater for viral RNA fragments by quantitative reverse transcription polymerase chain reaction (qRT-PCR) as a proxy of COVID-19 prevalence in the community. Most groups report low levels of viral RNA that often skirt the methods theoretical limits of detection and quantitation. Here, we demonstrate the presence of SARS-CoV-2 structural proteins in wastewater using traditional immunoblotting and quantitate them from wastewater solids using an immuno-linked PCR method called Multiplex Paired-antibody Amplified Detection (MPAD). In this longitudinal study, we corrected for stochastic variability inherent to wastewater-based epidemiology using multiple fecal content protein biomarkers. These normalized SARS-CoV-2 protein data correlated well with public health metrics. Our method of assaying SARS-CoV-2 protein from wastewater represents a promising and sensitive epidemiological tool to assess prevalence of fecally-shed pathogens in the community.

Wastewater-Based Epidemiology for Community Monitoring of SARS-CoV-2: Progress and Challenges

“…Compared to the RT-qPCR method for viral RNA, the MPAD method for the N-protein required fewer amplification cycles (Ct over 37 for RT-PCR vs 30 for MPAD on average). 107 Alternatively, Song et al has described the use of aptamers as affinity ligands which recognized the receptor binding domain of the S protein. They developed two DNA aptamers that are potential affinity ligands for developing diagnostic assays.…”

High-throughput wastewater analysis for substance use assessment in central New York during the COVID-19 pandemic

“…substances may serve as a proxy of SARS-CoV-2 RNA abundance in wastewater especially when evaluated against SARS-CoV-2 structural proteins. 102 In short, nontarget screening holds good promise as a hypothesis-generating method for prioritizing unknown substances as possible wastewater biomarkers of SARS-CoV-2 RNA prevalence, although gaps in metabolomics and informatics approaches should be addressed in order to identify the most relevant candidates for in-depth investigations.…”

Wastewater-based epidemiology for tracking COVID-19 trend and variants of concern in Ohio, United States

“…Our results agreed with the findings from other studies, which concluded that the PMMoV-normalized SARS-CoV-2 signal had lower background noise and showed the strongest correlation with active cases (57, 59). For future studies where normalization is required, combined markers of viral indicators, solid contents, and other volumetric parameters are recommended (60).…”