Author: Mohon, Abu Naser; Oberding, Lisa; Hundt, Jana; van Marle, Guido; Pabbaraju, Kanti; Berenger, Byron; Lisboa, Luiz; Griener, Thomas; Czub, Markus; Doolan, Cody; Servelitta, Venice; Chiu, Charles; Greninger, Alexander; Jerome, Keith; Pillai, Dylan R.
Title: Optimization and clinical validation of dual-target RT-LAMP for SARS-CoV-2 Cord-id: kftffes0 Document date: 2020_9_15
ID: kftffes0
Snippet: A 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 copies per reaction) as 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
Document: A 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 copies per reaction) as 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% to 99.96%) and NPA 100.00% (95% CI 93.84% to 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.
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