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

Optimization and clinical validation of dual-target RT-LAMP for SARS-CoV-2.
J Virol Methods. 2020 Sep 14;:113972
Authors: Mohon AN, Oberding L, Hundt J, van Marle G, Pabbaraju K, Berenger B, Lisboa L, Griener T, Czub M, Doolan C, Servelitta V, Chiu C, Greninger A, Jerome K, Pillai DR
Abstract
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.
PMID: 32941977 [PubMed - as supplied by publisher]

Source: Journal of Virological Methods - September 13, 2020 Category: Virology Authors: Mohon AN, Oberding L, Hundt J, van Marle G, Pabbaraju K, Berenger B, Lisboa L, Griener T, Czub M, Doolan C, Servelitta V, Chiu C, Greninger A, Jerome K, Pillai DR Tags: J Virol Methods Source Type: research