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

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 dem...

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Autores principales: Kim, Youngeun, Yaseen, Adam B., Kishi, Jocelyn Y., Hong, Fan, Saka, Sinem K., Sheng, Kuanwei, Gopalkrishnan, Nikhil, Schaus, Thomas E., Yin, Peng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7444299/
https://www.ncbi.nlm.nih.gov/pubmed/32839783
http://dx.doi.org/10.1101/2020.08.17.20177006
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author Kim, Youngeun
Yaseen, Adam B.
Kishi, Jocelyn Y.
Hong, Fan
Saka, Sinem K.
Sheng, Kuanwei
Gopalkrishnan, Nikhil
Schaus, Thomas E.
Yin, Peng
author_facet Kim, Youngeun
Yaseen, Adam B.
Kishi, Jocelyn Y.
Hong, Fan
Saka, Sinem K.
Sheng, Kuanwei
Gopalkrishnan, Nikhil
Schaus, Thomas E.
Yin, Peng
author_sort Kim, Youngeun
collection PubMed
description 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.
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spelling pubmed-74442992020-08-25 Single-strand RPA for rapid and sensitive detection of SARS-CoV-2 RNA Kim, Youngeun Yaseen, Adam B. Kishi, Jocelyn Y. Hong, Fan Saka, Sinem K. Sheng, Kuanwei Gopalkrishnan, Nikhil Schaus, Thomas E. Yin, Peng medRxiv Article 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. Cold Spring Harbor Laboratory 2020-10-25 /pmc/articles/PMC7444299/ /pubmed/32839783 http://dx.doi.org/10.1101/2020.08.17.20177006 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Kim, Youngeun
Yaseen, Adam B.
Kishi, Jocelyn Y.
Hong, Fan
Saka, Sinem K.
Sheng, Kuanwei
Gopalkrishnan, Nikhil
Schaus, Thomas E.
Yin, Peng
Single-strand RPA for rapid and sensitive detection of SARS-CoV-2 RNA
title Single-strand RPA for rapid and sensitive detection of SARS-CoV-2 RNA
title_full Single-strand RPA for rapid and sensitive detection of SARS-CoV-2 RNA
title_fullStr Single-strand RPA for rapid and sensitive detection of SARS-CoV-2 RNA
title_full_unstemmed Single-strand RPA for rapid and sensitive detection of SARS-CoV-2 RNA
title_short Single-strand RPA for rapid and sensitive detection of SARS-CoV-2 RNA
title_sort single-strand rpa for rapid and sensitive detection of sars-cov-2 rna
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7444299/
https://www.ncbi.nlm.nih.gov/pubmed/32839783
http://dx.doi.org/10.1101/2020.08.17.20177006
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