Stabilization of SARS-CoV-2 RNA in wastewater via rapid RNA extraction

Soroosh Torabi; Atena Amirsoleimani; Mohammad Dehghan Banadaki; William Dalton Strike; Alexus Rockward; Ann Noble; Matthew Liversedge; James W. Keck; Scott M. Berry

doi:10.1016/j.scitotenv.2023.162992

Stabilization of SARS-CoV-2 RNA in wastewater via rapid RNA extraction

Soroosh Torabi, Atena Amirsoleimani, Mohammad Dehghan Banadaki, William Dalton Strike, Alexus Rockward, Ann Noble, Matthew Liversedge, James W. Keck, Scott M. Berry

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

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.

Original language	English
Article number	162992
Journal	Science of the Total Environment
Volume	878
DOIs	https://doi.org/10.1016/j.scitotenv.2023.162992
State	Published - Jun 20 2023

Bibliographical note

Funding 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.

Funding 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 .

Publisher Copyright:
© 2023 Elsevier B.V.

Keywords

COVID-19
Degradation
RNA stabilization
Temperature
Wastewater

ASJC Scopus subject areas

Environmental Engineering
Environmental Chemistry
Waste Management and Disposal
Pollution

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.scitotenv.2023.162992

Cite this

@article{0db154dedb0748c990ddcaa9202372c5,

title = "Stabilization of SARS-CoV-2 RNA in wastewater via rapid RNA extraction",

abstract = "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.",

keywords = "COVID-19, Degradation, RNA stabilization, Temperature, Wastewater",

author = "Soroosh Torabi and Atena Amirsoleimani and {Dehghan Banadaki}, Mohammad and Strike, {William Dalton} and Alexus Rockward and Ann Noble and Matthew Liversedge and Keck, {James W.} and Berry, {Scott M.}",

note = "Funding 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. Funding 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 . Publisher Copyright: {\textcopyright} 2023 Elsevier B.V.",

year = "2023",

month = jun,

day = "20",

doi = "10.1016/j.scitotenv.2023.162992",

language = "English",

volume = "878",

}

TY - JOUR

T1 - Stabilization of SARS-CoV-2 RNA in wastewater via rapid RNA extraction

AU - Torabi, Soroosh

AU - Amirsoleimani, Atena

AU - Dehghan Banadaki, Mohammad

AU - Strike, William Dalton

AU - Rockward, Alexus

AU - Noble, Ann

AU - Liversedge, Matthew

AU - Keck, James W.

AU - Berry, Scott M.

N1 - Funding 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. Funding 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 . Publisher Copyright: © 2023 Elsevier B.V.

PY - 2023/6/20

Y1 - 2023/6/20

N2 - 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.

AB - 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.

KW - COVID-19

KW - Degradation

KW - RNA stabilization

KW - Temperature

KW - Wastewater

UR - http://www.scopus.com/inward/record.url?scp=85151013065&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85151013065&partnerID=8YFLogxK

U2 - 10.1016/j.scitotenv.2023.162992

DO - 10.1016/j.scitotenv.2023.162992

M3 - Article

C2 - 36948314

AN - SCOPUS:85151013065

SN - 0048-9697

VL - 878

JO - Science of the Total Environment

JF - Science of the Total Environment

M1 - 162992

ER -

Stabilization of SARS-CoV-2 RNA in wastewater via rapid RNA extraction

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this