Wastewater-based Epidemiology (WBE) has contributed to surveillance of SARS-CoV-2 in communities across the world. Both symptomatic and asymptomatic patients with COVID-19 can shed the virus through the gastrointestinal tract, enabling the quantification of the virus in stool and ultimately in wastewater (WW). Unfortunately, instability of SARS-CoV-2 RNA in wastewater limits the utility of WBE programs, particularly in remote/rural regions where reliable cold storage and/or rapid shipping may be unavailable. This study examined whether rapid SARS-CoV-2 RNA extraction on the day of sample collection could minimize degradation. Importantly, the extraction technology used in these experiments, termed exclusion-based sample preparation (ESP), is lightweight, portable, and electricity-free, making it suitable for implementation in remote settings. We demonstrated that immediate RNA extraction followed by ambient storage significantly increased the RNA half-life compared to raw wastewater samples stored at both 4 °C or ambient temperature. Given that RNA degradation negatively impacts both the sensitivity and precision of WBE measurements, efforts must be made to mitigate degradation in order to maximize the potential impact of WBE on public health.
|Journal||Science of the Total Environment|
|State||Published - Jun 20 2023|
Bibliographical noteFunding Information:
The work was supported by the National Institutes of Health (NIH) grants 1U01DA053903-01 and P30 ES026529 and the Centers for Disease Control and Prevention (CDC) contract BAA 75D301-20-R-68024. The following reagent was obtained through BEI Resources, NIAID, NIH: Genomic RNA from SARS-Related Coronavirus 2, Isolate USA/CA_CDC_5574/2020 (Lineage B.1.1.7), NR-55244, contributed by Centers for Disease Control and Prevention. The following reagent was deposited by the Centers for Disease Control and Prevention and obtained through BEI Resources, NIAID, NIH: SARS-Related Coronavirus 2, Isolate USA-WA1/2020, Heat Inactivated, NR-52286, Genomic RNA from SARS-Related Coronavirus 2, Isolate USA-CA4/2020, NR-52508. We thank Blazan Mijatovic, Cullen Hunter, and Savannah Tucker, field technicians at the University of Kentucky, for their assistance in collection of wastewater samples. Scott M. Berry has an ownership interest in Salus Discovery, LLC, which has licensed the ESP technology described in this work.
The work was supported by the National Institutes of Health (NIH) grants 1U01DA053903-01 and P30 ES026529 and the Centers for Disease Control and Prevention (CDC) contract BAA 75D301-20-R-68024 .
© 2023 Elsevier B.V.
- RNA stabilization
ASJC Scopus subject areas
- Environmental Engineering
- Environmental Chemistry
- Waste Management and Disposal