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Preliminary optimisation of a simplified sample preparation method to permit direct detection of SARS-CoV-2 within saliva samples using reverse-transcription loop-mediated isothermal amplification (RT-LAMP)

Preliminary optimisation of a simplified sample preparation method to permit direct detection of SARS-CoV-2 within saliva samples using reverse-transcription loop-mediated isothermal amplification (RT-LAMP)
Preliminary optimisation of a simplified sample preparation method to permit direct detection of SARS-CoV-2 within saliva samples using reverse-transcription loop-mediated isothermal amplification (RT-LAMP)
We describe the optimisation of a simplified sample preparation method which permits rapid and direct detection of SARS-CoV-2 RNA within saliva, using reverse-transcription loop-mediated isothermal amplification (RT-LAMP). Treatment of saliva samples prior to RT-LAMP by dilution 1:1 in Mucolyse™, followed by dilution in 10 % (w/v) Chelex© 100 Resin and a 98 °C heat step for 2 min enabled detection of SARS-CoV-2 RNA in positive saliva samples. Using RT-LAMP, SARS-CoV-2 RNA was detected in as little as 05:43 min, with no amplification detected in 3097 real-time reverse transcription PCR (rRT-PCR) negative saliva samples from staff tested within a service evaluation study, or for other respiratory pathogens tested (n = 22). Saliva samples can be collected non-invasively, without the need for skilled staff and can be obtained from both healthcare and home settings. Critically, this approach overcomes the requirement for, and validation of, different swabs and the global bottleneck in obtaining access to extraction robots and reagents to enable molecular testing by rRT-PCR. Such testing opens the possibility of public health approaches for effective intervention during the COVID-19 pandemic through regular SARS-CoV-2 testing at a population scale, combined with isolation and contact tracing.
0166-0934
Howson, Emma L. A.
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Kidd, Stephen P.
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Armson, Bryony
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Goring, Alice
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Sawyer, Jason
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Cassar, Claire
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Cross, David
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Lewis, Tom
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Hockey, Jess
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Rivers, Samantha
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Cawthraw, Saira
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Banyard, Ashley
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Anderson, Paul
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Rahou, Sabah
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Andreou, Michael
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Morant, Nick
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Clark, Duncan
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Walsh, Charlotte
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Laxman, Shailen
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Houghton, Rebecca
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Slater-Jefferies, Joanne
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Costello, Paula
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Brown, Ian
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Cortes, Nicholas
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Godfrey, Keith
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Fowler, Veronica L.
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Howson, Emma L. A.
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Kidd, Stephen P.
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Armson, Bryony
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Goring, Alice
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Sawyer, Jason
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Cassar, Claire
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Cross, David
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Lewis, Tom
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Hockey, Jess
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Rivers, Samantha
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Cawthraw, Saira
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Banyard, Ashley
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Anderson, Paul
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Rahou, Sabah
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Andreou, Michael
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Morant, Nick
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Clark, Duncan
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Walsh, Charlotte
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Laxman, Shailen
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Houghton, Rebecca
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Slater-Jefferies, Joanne
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Costello, Paula
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Brown, Ian
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Cortes, Nicholas
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Godfrey, Keith
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Fowler, Veronica L.
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Howson, Emma L. A., Kidd, Stephen P., Armson, Bryony, Goring, Alice, Sawyer, Jason, Cassar, Claire, Cross, David, Lewis, Tom, Hockey, Jess, Rivers, Samantha, Cawthraw, Saira, Banyard, Ashley, Anderson, Paul, Rahou, Sabah, Andreou, Michael, Morant, Nick, Clark, Duncan, Walsh, Charlotte, Laxman, Shailen, Houghton, Rebecca, Slater-Jefferies, Joanne, Costello, Paula, Brown, Ian, Cortes, Nicholas, Godfrey, Keith and Fowler, Veronica L. (2020) Preliminary optimisation of a simplified sample preparation method to permit direct detection of SARS-CoV-2 within saliva samples using reverse-transcription loop-mediated isothermal amplification (RT-LAMP). Journal of Virological Methods, [114048]. (doi:10.1016/j.jviromet.2020.114048).

Record type: Article

Abstract

We describe the optimisation of a simplified sample preparation method which permits rapid and direct detection of SARS-CoV-2 RNA within saliva, using reverse-transcription loop-mediated isothermal amplification (RT-LAMP). Treatment of saliva samples prior to RT-LAMP by dilution 1:1 in Mucolyse™, followed by dilution in 10 % (w/v) Chelex© 100 Resin and a 98 °C heat step for 2 min enabled detection of SARS-CoV-2 RNA in positive saliva samples. Using RT-LAMP, SARS-CoV-2 RNA was detected in as little as 05:43 min, with no amplification detected in 3097 real-time reverse transcription PCR (rRT-PCR) negative saliva samples from staff tested within a service evaluation study, or for other respiratory pathogens tested (n = 22). Saliva samples can be collected non-invasively, without the need for skilled staff and can be obtained from both healthcare and home settings. Critically, this approach overcomes the requirement for, and validation of, different swabs and the global bottleneck in obtaining access to extraction robots and reagents to enable molecular testing by rRT-PCR. Such testing opens the possibility of public health approaches for effective intervention during the COVID-19 pandemic through regular SARS-CoV-2 testing at a population scale, combined with isolation and contact tracing.

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Saliva Direct RT-LAMP rapid communication_corrections - Accepted Manuscript
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Saliva Direct RT-LAMP rapid communication_tables_corr
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More information

Accepted/In Press date: 12 December 2020
e-pub ahead of print date: 20 December 2020

Identifiers

Local EPrints ID: 445959
URI: http://eprints.soton.ac.uk/id/eprint/445959
ISSN: 0166-0934
PURE UUID: 2c514df2-8470-4c86-8d9e-b4e3d50fe31a
ORCID for Joanne Slater-Jefferies: ORCID iD orcid.org/0000-0001-8325-1320
ORCID for Keith Godfrey: ORCID iD orcid.org/0000-0002-4643-0618

Catalogue record

Date deposited: 15 Jan 2021 17:31
Last modified: 01 May 2021 01:40

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Contributors

Author: Emma L. A. Howson
Author: Stephen P. Kidd
Author: Bryony Armson
Author: Alice Goring
Author: Jason Sawyer
Author: Claire Cassar
Author: David Cross
Author: Tom Lewis
Author: Jess Hockey
Author: Samantha Rivers
Author: Saira Cawthraw
Author: Ashley Banyard
Author: Paul Anderson
Author: Sabah Rahou
Author: Michael Andreou
Author: Nick Morant
Author: Duncan Clark
Author: Charlotte Walsh
Author: Shailen Laxman
Author: Rebecca Houghton
Author: Paula Costello
Author: Ian Brown
Author: Nicholas Cortes
Author: Keith Godfrey ORCID iD
Author: Veronica L. Fowler

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