Improved recovery of SARS-CoV-2 from wastewater through application of RNA and DNA stabilising agents

Stephen H Bell; Danielle M Allen; Marina I Reyne; Jonathan F W Lock; Arthur Fitzgerald; Ashley Levickas; Andrew J Lee; Connor G G Bamford; Deirdre F Gilpin; John W McGrath

doi:10.1093/lambio/ovad047

Improved recovery of SARS-CoV-2 from wastewater through application of RNA and DNA stabilising agents

Stephen H Bell^*
, Danielle M Allen
, Marina I Reyne
, Jonathan F W Lock
, Arthur Fitzgerald
, Ashley Levickas
, Andrew J Lee
, Connor G G Bamford
, Deirdre F Gilpin
, John W McGrath

^*Corresponding author for this work

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

5 Citations (Scopus)

64 Downloads (Pure)

Abstract

Wastewater Based Epidemiology (WBE) has become an integral part of the public health effort to track the levels of SARS-CoV-2 within communities. Detection of SARS-CoV-2 in wastewater can be challenging due to relatively low levels of virus within the sample. The wastewater matrix is also comprised of commercial and domestically derived contaminants, as well as RNases, all of which can adversely affect RT-qPCR analysis. To improve SARS-CoV-2 detection within wastewater samples we investigated both the effect of template dilution (as a means to reduce RT-qPCR inhibition) and sample stabilisation via addition of DNA/RNA Shield™ and/or RNA Later™ (to prevent RNA degradation via RNases) as a means to improve viral fragment detection. Using both methodologies a significant improvement in SARS-CoV-2 detection from wastewater samples was observed. No adverse effects of stabilising agent addition on downstream Next Generation Sequencing workflows were detected.

Original language	English
Journal	Letters in Applied Microbiology
Early online date	03 May 2023
DOIs	https://doi.org/10.1093/lambio/ovad047
Publication status	Early online date - 03 May 2023

Bibliographical note

UN SDGs

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

SDG 3 Good Health and Well-being

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10.1093/lambio/ovad047Licence: Unspecified

Improved recovery of SARS-CoV-2 from wastewater through application of RNA and DNA stabilising agents
Copyright 2023, the Authors. This work is made available online in accordance with the publisher’s policies. Please refer to any applicable terms of use of the publisher.
Accepted author manuscript, 1.18 MBLicence: Unspecified

Cite this

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title = "Improved recovery of SARS-CoV-2 from wastewater through application of RNA and DNA stabilising agents",

abstract = "Wastewater Based Epidemiology (WBE) has become an integral part of the public health effort to track the levels of SARS-CoV-2 within communities. Detection of SARS-CoV-2 in wastewater can be challenging due to relatively low levels of virus within the sample. The wastewater matrix is also comprised of commercial and domestically derived contaminants, as well as RNases, all of which can adversely affect RT-qPCR analysis. To improve SARS-CoV-2 detection within wastewater samples we investigated both the effect of template dilution (as a means to reduce RT-qPCR inhibition) and sample stabilisation via addition of DNA/RNA Shield{\texttrademark} and/or RNA Later{\texttrademark} (to prevent RNA degradation via RNases) as a means to improve viral fragment detection. Using both methodologies a significant improvement in SARS-CoV-2 detection from wastewater samples was observed. No adverse effects of stabilising agent addition on downstream Next Generation Sequencing workflows were detected.",

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AU - Bell, Stephen H

AU - Allen, Danielle M

AU - Reyne, Marina I

AU - Lock, Jonathan F W

AU - Fitzgerald, Arthur

AU - Levickas, Ashley

AU - Lee, Andrew J

AU - Bamford, Connor G G

AU - Gilpin, Deirdre F

AU - McGrath, John W

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N2 - Wastewater Based Epidemiology (WBE) has become an integral part of the public health effort to track the levels of SARS-CoV-2 within communities. Detection of SARS-CoV-2 in wastewater can be challenging due to relatively low levels of virus within the sample. The wastewater matrix is also comprised of commercial and domestically derived contaminants, as well as RNases, all of which can adversely affect RT-qPCR analysis. To improve SARS-CoV-2 detection within wastewater samples we investigated both the effect of template dilution (as a means to reduce RT-qPCR inhibition) and sample stabilisation via addition of DNA/RNA Shield™ and/or RNA Later™ (to prevent RNA degradation via RNases) as a means to improve viral fragment detection. Using both methodologies a significant improvement in SARS-CoV-2 detection from wastewater samples was observed. No adverse effects of stabilising agent addition on downstream Next Generation Sequencing workflows were detected.

AB - Wastewater Based Epidemiology (WBE) has become an integral part of the public health effort to track the levels of SARS-CoV-2 within communities. Detection of SARS-CoV-2 in wastewater can be challenging due to relatively low levels of virus within the sample. The wastewater matrix is also comprised of commercial and domestically derived contaminants, as well as RNases, all of which can adversely affect RT-qPCR analysis. To improve SARS-CoV-2 detection within wastewater samples we investigated both the effect of template dilution (as a means to reduce RT-qPCR inhibition) and sample stabilisation via addition of DNA/RNA Shield™ and/or RNA Later™ (to prevent RNA degradation via RNases) as a means to improve viral fragment detection. Using both methodologies a significant improvement in SARS-CoV-2 detection from wastewater samples was observed. No adverse effects of stabilising agent addition on downstream Next Generation Sequencing workflows were detected.

U2 - 10.1093/lambio/ovad047

DO - 10.1093/lambio/ovad047

M3 - Article

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SN - 0266-8254

JO - Letters in Applied Microbiology

JF - Letters in Applied Microbiology

ER -

Improved recovery of SARS-CoV-2 from wastewater through application of RNA and DNA stabilising agents

Abstract

Bibliographical note

UN SDGs

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Cite this