• J. Virol. Methods · Dec 2020

    Optimization and clinical validation of dual-target RT-LAMP for SARS-CoV-2.

    • Abu Naser Mohon, Lisa Oberding, Jana Hundt, Guido van Marle, Kanti Pabbaraju, Byron M Berenger, Luiz Lisboa, Thomas Griener, Markus Czub, Cody Doolan, Venice Servellita, Charles Y Chiu, Alexander L Greninger, Keith R Jerome, and Dylan R Pillai.
    • Department of Microbiology, Immunology, and Infectious Diseases, University of Calgary, AB, Canada.
    • J. Virol. Methods. 2020 Dec 1; 286: 113972.

    AbstractA novel reverse-transcriptase loop mediated amplification (RT-LAMP) method targeting genes encoding the Spike (S) protein and RNA-dependent RNA polymerase (RdRP) of SARS-CoV-2 has been developed. The LAMP assay achieves a comparable limit of detection (25-50 copies per reaction) to commonly used RT-PCR protocols using clinical samples quantified by digital droplet PCR. Precision, cross-reactivity, inclusivity, and limit of detection studies were performed according to regulatory standards. Clinical validation of dual-target RT-LAMP (S and RdRP gene) achieved a PPA of 98.48 % (95 % CI 91.84%-99.96%) and NPA 100.00 % (95 % CI 93.84%-100.00%) based on the E gene and N2 gene reference RT-PCR methods. The method has implications for development of point of care technology using isothermal amplification.Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.

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