Author: Mohon, A. N.; Hundt, J.; van Marle, G.; Pabbaraju, K.; Berenger, B.; Griener, T.; Lisboa, L.; Church, D.; Czub, M.; Greninger, A.; Jerome, K.; Doolan, C.; Pillai, D. R.
Title: Development and validation of direct RT-LAMP for SARS-CoV-2 Cord-id: 6uci7nu4 Document date: 2020_5_4
ID: 6uci7nu4
Snippet: 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 the same limit of detection as commonly used RT-PCR protocols based on artificial targets, recombinant Sindbis virus, and clinical samples. Clinical validation of single target LAMP (N=108) showed a positive percent agreement (PPA) of 33/34 (97.1%) and negative percent agreement (NPA) o
Document: 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 the same limit of detection as commonly used RT-PCR protocols based on artificial targets, recombinant Sindbis virus, and clinical samples. Clinical validation of single target LAMP (N=108) showed a positive percent agreement (PPA) of 33/34 (97.1%) and negative percent agreement (NPA) of 73/74 (98.6%) compared to reference RT-PCR. Dual target RT-LAMP achieved a PPA of 11/11 (100%) and NPA 13/13 (100%) when including discrepant samples. The assay can be performed without a formal extraction procedure, with lyophilized reagents that do need cold chain, and is amenable to point-of-care application with visual detection.
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