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Single-strand RPA for rapid and sensitive detection of SARS-CoV-2 RNA.

ABSTRACT: We report the single-strand Recombinase Polymerase Amplification (ssRPA) method, which merges the fast, isothermal amplification of RPA with subsequent rapid conversion of the double-strand DNA amplicon to single strands, and hence enables facile hybridization-based, high-specificity readout. We demonstrate the utility of ssRPA for sensitive and rapid (4 copies per 50 μL reaction within 10 min, or 8 copies within 8 min) visual detection of SARS-CoV-2 RNA spiked samples, as well as clinical saliva and nasopharyngeal swabs in VTM or water, on lateral flow devices. The ssRPA method promises rapid, sensitive, and accessible RNA detection to facilitate mass testing in the COVID-19 pandemic.

SUBMITTER: Kim Y

PROVIDER: S-EPMC7444299 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Single-strand RPA for rapid and sensitive detection of SARS-CoV-2 RNA.

Kim Youngeun Y Yaseen Adam B AB Kishi Jocelyn Y JY Hong Fan F Saka Sinem K SK Sheng Kuanwei K Gopalkrishnan Nikhil N Schaus Thomas E TE Yin Peng P

medRxiv : the preprint server for health sciences 20201025

We report the single-strand Recombinase Polymerase Amplification (ssRPA) method, which merges the fast, isothermal amplification of RPA with subsequent rapid conversion of the double-strand DNA amplicon to single strands, and hence enables facile hybridization-based, high-specificity readout. We demonstrate the utility of ssRPA for sensitive and rapid (4 copies per 50 μL reaction within 10 min, or 8 copies within 8 min) visual detection of SARS-CoV-2 RNA spiked samples, as well as clinical saliv ...[more]

PMID: 32839783

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