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).
J Virol Methods
; 289: 114048, 2021 03.
Article
in English
| MEDLINE | ID: covidwho-988686
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 11 in Mucolyse™, followed by dilution in 10 % (w/v) Chelex© 100 Resin and a 98⯰C heat step for 2â¯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 0543â¯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 (nâ¯=â¯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.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Saliva
/
Specimen Handling
/
Nucleic Acid Amplification Techniques
/
Molecular Diagnostic Techniques
/
COVID-19 Testing
/
SARS-CoV-2
/
COVID-19
Type of study:
Diagnostic study
/
Experimental Studies
/
Prognostic study
Limits:
Humans
Language:
English
Journal:
J Virol Methods
Year:
2021
Document Type:
Article
Affiliation country:
J.jviromet.2020.114048
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