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Development and validation of direct RT-LAMP for SARS-CoV-2

2020; Cold Spring Harbor Laboratory; Linguagem: Inglês

10.1101/2020.04.29.20075747

Abu Naser Mohon, Jana Hundt, Guido van Marle, Kanti Pabbaraju, Byron M. Berenger, Thomas P. Griener, Luiz F. Lisboa, Deirdre L. Church, Markus Czub, Alexander L. Greninger, Keith R. Jerome, Cody Doolan, Dylan R. Pillai,

Advanced biosensing and bioanalysis techniques

Abstract We have developed a 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. The LAMP assay achieves comparable limit of detection as commonly used RT-PCR protocols based on artificial targets, recombinant Sindbis virus, and clinical samples. Clinical validation of single-target (S gene) LAMP (N=120) showed a positive percent agreement (PPA) of 41/42 (97.62%) and negative percent agreement (NPA) of 77/78 (98.72%) compared to reference RT-PCR. Dual-target RT-LAMP (S and RdRP gene) achieved a PPA of 44/48 (91.97%) and NPA 72/72 (100%) when including discrepant samples. The assay can be performed without a formal extraction procedure, with lyophilized reagents which do need cold chain, and is amenable to point-of-care application with visual detection.