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A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing

RT-qPCR-based diagnostic tests play important roles in combating virus-caused pandemics such as Covid-19. However, their dependence on sophisticated equipment and the associated costs often limits their widespread use. Loop-mediated isothermal amplification after reverse transcription (RT-LAMP) is a...

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Autores principales: Kellner, Max J., Ross, James J., Schnabl, Jakob, Dekens, Marcus P. S., Matl, Martin, Heinen, Robert, Grishkovskaya, Irina, Bauer, Benedikt, Stadlmann, Johannes, Menéndez-Arias, Luis, Straw, Andrew D., Fritsche-Polanz, Robert, Traugott, Marianna, Seitz, Tamara, Zoufaly, Alexander, Födinger, Manuela, Wenisch, Christoph, Zuber, Johannes, Pauli, Andrea, Brennecke, Julius
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9011764/
https://www.ncbi.nlm.nih.gov/pubmed/35433827
http://dx.doi.org/10.3389/fmolb.2022.801309
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author Kellner, Max J.
Ross, James J.
Schnabl, Jakob
Dekens, Marcus P. S.
Matl, Martin
Heinen, Robert
Grishkovskaya, Irina
Bauer, Benedikt
Stadlmann, Johannes
Menéndez-Arias, Luis
Straw, Andrew D.
Fritsche-Polanz, Robert
Traugott, Marianna
Seitz, Tamara
Zoufaly, Alexander
Födinger, Manuela
Wenisch, Christoph
Zuber, Johannes
Pauli, Andrea
Brennecke, Julius
author_facet Kellner, Max J.
Ross, James J.
Schnabl, Jakob
Dekens, Marcus P. S.
Matl, Martin
Heinen, Robert
Grishkovskaya, Irina
Bauer, Benedikt
Stadlmann, Johannes
Menéndez-Arias, Luis
Straw, Andrew D.
Fritsche-Polanz, Robert
Traugott, Marianna
Seitz, Tamara
Zoufaly, Alexander
Födinger, Manuela
Wenisch, Christoph
Zuber, Johannes
Pauli, Andrea
Brennecke, Julius
author_sort Kellner, Max J.
collection PubMed
description RT-qPCR-based diagnostic tests play important roles in combating virus-caused pandemics such as Covid-19. However, their dependence on sophisticated equipment and the associated costs often limits their widespread use. Loop-mediated isothermal amplification after reverse transcription (RT-LAMP) is an alternative nucleic acid detection method that overcomes these limitations. Here, we present a rapid, robust, and sensitive RT-LAMP-based SARS-CoV-2 detection assay. Our 40-min procedure bypasses the RNA isolation step, is insensitive to carryover contamination, and uses a colorimetric readout that enables robust SARS-CoV-2 detection from various sample types. Based on this assay, we have increased sensitivity and scalability by adding a nucleic acid enrichment step (Bead-LAMP), developed a version for home testing (HomeDip-LAMP), and identified open-source RT-LAMP enzymes that can be produced in any molecular biology laboratory. On a dedicated website, rtlamp.org (DOI: 10.5281/zenodo.6033689), we provide detailed protocols and videos. Our optimized, general-purpose RT-LAMP assay is an important step toward population-scale SARS-CoV-2 testing.
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spelling pubmed-90117642022-04-16 A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing Kellner, Max J. Ross, James J. Schnabl, Jakob Dekens, Marcus P. S. Matl, Martin Heinen, Robert Grishkovskaya, Irina Bauer, Benedikt Stadlmann, Johannes Menéndez-Arias, Luis Straw, Andrew D. Fritsche-Polanz, Robert Traugott, Marianna Seitz, Tamara Zoufaly, Alexander Födinger, Manuela Wenisch, Christoph Zuber, Johannes Pauli, Andrea Brennecke, Julius Front Mol Biosci Molecular Biosciences RT-qPCR-based diagnostic tests play important roles in combating virus-caused pandemics such as Covid-19. However, their dependence on sophisticated equipment and the associated costs often limits their widespread use. Loop-mediated isothermal amplification after reverse transcription (RT-LAMP) is an alternative nucleic acid detection method that overcomes these limitations. Here, we present a rapid, robust, and sensitive RT-LAMP-based SARS-CoV-2 detection assay. Our 40-min procedure bypasses the RNA isolation step, is insensitive to carryover contamination, and uses a colorimetric readout that enables robust SARS-CoV-2 detection from various sample types. Based on this assay, we have increased sensitivity and scalability by adding a nucleic acid enrichment step (Bead-LAMP), developed a version for home testing (HomeDip-LAMP), and identified open-source RT-LAMP enzymes that can be produced in any molecular biology laboratory. On a dedicated website, rtlamp.org (DOI: 10.5281/zenodo.6033689), we provide detailed protocols and videos. Our optimized, general-purpose RT-LAMP assay is an important step toward population-scale SARS-CoV-2 testing. Frontiers Media S.A. 2022-04-01 /pmc/articles/PMC9011764/ /pubmed/35433827 http://dx.doi.org/10.3389/fmolb.2022.801309 Text en Copyright © 2022 Kellner, Ross, Schnabl, Dekens, Matl, Heinen, Grishkovskaya, Bauer, Stadlmann, Menéndez-Arias, Straw, Fritsche-Polanz, Traugott, Seitz, Zoufaly, Födinger, Wenisch, Zuber, Vienna COVID-19 Detection Initiative (VCDI), Pauli and Brennecke. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Kellner, Max J.
Ross, James J.
Schnabl, Jakob
Dekens, Marcus P. S.
Matl, Martin
Heinen, Robert
Grishkovskaya, Irina
Bauer, Benedikt
Stadlmann, Johannes
Menéndez-Arias, Luis
Straw, Andrew D.
Fritsche-Polanz, Robert
Traugott, Marianna
Seitz, Tamara
Zoufaly, Alexander
Födinger, Manuela
Wenisch, Christoph
Zuber, Johannes
Pauli, Andrea
Brennecke, Julius
A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing
title A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing
title_full A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing
title_fullStr A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing
title_full_unstemmed A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing
title_short A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing
title_sort rapid, highly sensitive and open-access sars-cov-2 detection assay for laboratory and home testing
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9011764/
https://www.ncbi.nlm.nih.gov/pubmed/35433827
http://dx.doi.org/10.3389/fmolb.2022.801309
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