Journal article
Clinical validation of optimised RT-LAMP for the diagnosis of SARS-CoV-2 infection
- Abstract:
- We have optimised a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the detection of SARS-CoV-2 from extracted RNA for clinical application. We improved the stability and reliability of the RT-LAMP assay by the addition of a temperature-dependent switch oligonucleotide to reduce self- or off-target amplification. We then developed freeze-dried master mix for single step RT-LAMP reaction, simplifying the operation for end users and improving long-term storage and transportation. The assay can detect as low as 13 copies of SARS-CoV2 RNA per reaction (25-μL). Cross reactivity with other human coronaviruses was not observed. We have applied the new RT-LAMP assay for testing clinical extracted RNA samples extracted from swabs of 72 patients in the UK and 126 samples from Greece and demonstrated the overall sensitivity of 90.2% (95% CI 83.8–94.7%) and specificity of 92.4% (95% CI 83.2–97.5%). Among 115 positive samples which Ct values were less than 34, the RT-LAMP assay was able to detect 110 of them with 95.6% sensitivity. The specificity was 100% when RNA elution used RNase-free water. The outcome of RT-LAMP can be reported by both colorimetric detection and quantifiable fluorescent reading. Objective measures with a digitized reading data flow would allow for the sharing of results for local or national surveillance.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, 5.6MB, Terms of use)
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- Publisher copy:
- 10.1038/s41598-021-95607-1
Authors
+ EPSRC and Oxford University
More from this funder
- Funding agency for:
- Andersson, M
- Cui, Z
- Huang, W
- Programme:
- COVID-19 Research Response Fund
- Publisher:
- Springer Nature
- Journal:
- Scientific Reports More from this journal
- Volume:
- 11
- Issue:
- 1
- Article number:
- 16193
- Publication date:
- 2021-08-10
- Acceptance date:
- 2021-07-21
- DOI:
- EISSN:
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2045-2322
- Language:
-
English
- Keywords:
- Pubs id:
-
1178371
- Local pid:
-
pubs:1178371
- Deposit date:
-
2021-05-25
Terms of use
- Copyright holder:
- Lim et al.
- Copyright date:
- 2021
- Rights statement:
- © 2021 The Authors. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
- Licence:
- CC Attribution (CC BY)
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