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A highly effective reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for the rapid detection of SARS-CoV-2 infection

Research output: Contribution to journalArticlepeer-review

  • Veronica L. Fowler
  • Bryony Armson
  • Jose L. Gonzales
  • Emma L. Wise
  • Emma L.A. Howson
  • Zoe Vincent-Mistiaen
  • Connor J. Maltby
  • Seden Grippon
  • Simon Munro
  • Lisa Jones
  • Tom Holmes
  • Claire Tillyer
  • Joanne Elwell
  • Amy Sowood
  • Oliver De Peyer
  • Sophie Dixon
  • Thomas Hatcher
  • Helen Patrick
  • Shailen Laxman
  • Charlotte Walsh
  • Michael Andreou
  • Nick Morant
  • Duncan Clark
  • Nathan Moore
  • Rebecca Houghton
  • Nicholas Cortes
  • Stephen P. Kidd
The COVID-19 pandemic has illustrated the importance of simple, rapid and accurate diagnostic testing. This study describes the validation of a new rapid SARS-CoV-2 RT-LAMP assay for use on extracted RNA or directly from swab offering an alternative diagnostic pathway that does not rely on traditional reagents that are often in short supply during a pandemic.

Analytical specificity (ASp) of this new RT-LAMP assay was 100% and analytical sensitivity (ASe) was between 1 × 101 and 1 × 102 copies per reaction when using a synthetic DNA target. The overall diagnostic sensitivity (DSe) and specificity (DSp) of RNA RT-LAMP was 97% and 99% respectively, relative to the standard of care rRT-PCR. When a CT cut-off of 33 was employed, above which increasingly evidence suggests there is a low risk of patients shedding infectious virus, the diagnostic sensitivity was 100%. The DSe and DSp of Direct RT-LAMP (that does not require RNA extraction) was 67% and 97%, respectively. When setting CT cut-offs of ≤33 and ≤25, the DSe increased to 75% and 100%, respectively, time from swab-to-result, CT < 25, was < 15 minutes.

We propose that RNA RT-LAMP could replace rRT-PCR where there is a need for increased sample throughput and Direct RT-LAMP as a near-patient screening tool to rapidly identify highly contagious individuals within emergency departments and a care homes during times of increased disease prevalence ensuring negative results still get laboratory confirmation.
Original languageEnglish
JournalJournal of Infection
Early online date30 Nov 2020
DOIs
Publication statusEarly online - 30 Nov 2020

Documents

  • A highly effective reverse-transcription Post-print

    Accepted author manuscript (Post-print), 446 KB, PDF document

    Due to publisher’s copyright restrictions, this document is not freely available to download from this website until: 30/11/21

    Licence: CC BY-NC-ND

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