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™, fo...

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Bibliographic Details
Published in:	Journal of virological methods Vol. 289; p. 114048
Main Authors:	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 M., Fowler, Veronica L.
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 01-03-2021
Subjects:	COVID-19 COVID-19 - diagnosis COVID-19 Testing - methods Direct detection Humans Molecular Diagnostic Techniques - methods Near patient testing Nucleic Acid Amplification Techniques - methods Rapid diagnostics RNA, Viral - analysis RT-LAMP Saliva Saliva - virology SARS-CoV-2 SARS-CoV-2 - isolation & purification Short Communication Specimen Handling - methods COVID-19 RT-LAMP SARS-CoV-2 Direct detection Near patient testing Saliva Rapid diagnostics
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Joint first authors.
ISSN:	0166-0934 1879-0984
DOI:	10.1016/j.jviromet.2020.114048