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A rapid, highly sensitive and open-access SARS-CoV-2 detection assay for laboratory and home testing

Max J. Kellner, James J. Ross, Jakob Schnabl, Marcus P.S. Dekens, Robert Heinen, Irina Grishkovskaya, Benedikt Bauer, Johannes Stadlmann, Luis Menéndez-Arias, Robert Fritsche-Polanz, Marianna Traugott, Tamara Seitz, Alexander Zoufaly, Manuela Födinger, Christoph Wenisch, Johannes Zuber, Vienna Covid-19 Diagnostics Initiative (VCDI), Andrea Pauli, Julius Brennecke
doi: https://doi.org/10.1101/2020.06.23.166397
Max J. Kellner
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Campus-Vienna-Biocenter 1, 1030 Vienna, Austria
2Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Dr. Bohr-Gasse 3, 1030 Vienna, Austria
3MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK
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  • For correspondence: mkellner@mrc-lmb.cam.ac.uk andrea.pauli@imp.ac.at julius.brennecke@imba.oeaw.ac.at
James J. Ross
2Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Dr. Bohr-Gasse 3, 1030 Vienna, Austria
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Jakob Schnabl
2Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Dr. Bohr-Gasse 3, 1030 Vienna, Austria
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Marcus P.S. Dekens
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Campus-Vienna-Biocenter 1, 1030 Vienna, Austria
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Robert Heinen
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Campus-Vienna-Biocenter 1, 1030 Vienna, Austria
2Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Dr. Bohr-Gasse 3, 1030 Vienna, Austria
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Irina Grishkovskaya
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Campus-Vienna-Biocenter 1, 1030 Vienna, Austria
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Benedikt Bauer
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Campus-Vienna-Biocenter 1, 1030 Vienna, Austria
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Johannes Stadlmann
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Campus-Vienna-Biocenter 1, 1030 Vienna, Austria
2Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Dr. Bohr-Gasse 3, 1030 Vienna, Austria
4Department of Chemistry, University of Natural Resources and Life Sciences, Vienna; Muthgasse 18, 1190 Vienna, Austria
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Luis Menéndez-Arias
5Centro de Biología Molecular ‘Severo Ochoa’ (Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid), c/ Nicolás Cabrera 1, Campus de Cantoblanco-UAM, 28049 Madrid, Spain
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Robert Fritsche-Polanz
6Institute of Laboratory Diagnostics, Klinik Favoriten, 1100 Vienna, Austria
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Marianna Traugott
74th Medical Department with Infectious Diseases and Tropical Medicine, Klinik Favoriten, 1100 Vienna, Austria
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Tamara Seitz
74th Medical Department with Infectious Diseases and Tropical Medicine, Klinik Favoriten, 1100 Vienna, Austria
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Alexander Zoufaly
74th Medical Department with Infectious Diseases and Tropical Medicine, Klinik Favoriten, 1100 Vienna, Austria
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Manuela Födinger
6Institute of Laboratory Diagnostics, Klinik Favoriten, 1100 Vienna, Austria
8Sigmund Freud Private University, 1020 Vienna, Austria
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Christoph Wenisch
74th Medical Department with Infectious Diseases and Tropical Medicine, Klinik Favoriten, 1100 Vienna, Austria
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Johannes Zuber
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Campus-Vienna-Biocenter 1, 1030 Vienna, Austria
9Medical University of Vienna, Vienna BioCenter (VBC), 1030 Vienna, Austria
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Andrea Pauli
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Campus-Vienna-Biocenter 1, 1030 Vienna, Austria
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  • For correspondence: mkellner@mrc-lmb.cam.ac.uk andrea.pauli@imp.ac.at julius.brennecke@imba.oeaw.ac.at
Julius Brennecke
2Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Dr. Bohr-Gasse 3, 1030 Vienna, Austria
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  • For correspondence: mkellner@mrc-lmb.cam.ac.uk andrea.pauli@imp.ac.at julius.brennecke@imba.oeaw.ac.at
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Abstract

Global efforts to combat the Covid-19 pandemic caused by the beta coronavirus SARS-CoV-2 are currently based on RT-qPCR-based diagnostic tests. However, their high cost, moderate throughput and reliance on sophisticated equipment limit widespread implementation. Loop-mediated isothermal amplification after reverse transcription (RT-LAMP) is an alternative detection method that has the potential to overcome these limitations. Here we present a rapid, robust, highly sensitive and versatile RT-LAMP based SARS-CoV-2 detection assay. Our forty-minute procedure bypasses a dedicated RNA isolation step, is insensitive to carry-over contamination, and uses a hydroxynaphthol blue (HNB)-based colorimetric readout, which allows robust SARS-CoV-2 detection from various sample types. Based on this assay we have substantially increased sensitivity and scalability by a simple nucleic acid enrichment step (bead-LAMP), established a pipette-free version for home testing (HomeDip-LAMP), and developed a version with open source enzymes that could be produced in any molecular biology setting. Our advanced, universally applicable RT-LAMP assay is a major step towards population-scale SARS-CoV-2 testing.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted July 23, 2020.
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A rapid, highly sensitive and open-access SARS-CoV-2 detection assay for laboratory and home testing
Max J. Kellner, James J. Ross, Jakob Schnabl, Marcus P.S. Dekens, Robert Heinen, Irina Grishkovskaya, Benedikt Bauer, Johannes Stadlmann, Luis Menéndez-Arias, Robert Fritsche-Polanz, Marianna Traugott, Tamara Seitz, Alexander Zoufaly, Manuela Födinger, Christoph Wenisch, Johannes Zuber, Vienna Covid-19 Diagnostics Initiative (VCDI), Andrea Pauli, Julius Brennecke
bioRxiv 2020.06.23.166397; doi: https://doi.org/10.1101/2020.06.23.166397
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A rapid, highly sensitive and open-access SARS-CoV-2 detection assay for laboratory and home testing
Max J. Kellner, James J. Ross, Jakob Schnabl, Marcus P.S. Dekens, Robert Heinen, Irina Grishkovskaya, Benedikt Bauer, Johannes Stadlmann, Luis Menéndez-Arias, Robert Fritsche-Polanz, Marianna Traugott, Tamara Seitz, Alexander Zoufaly, Manuela Födinger, Christoph Wenisch, Johannes Zuber, Vienna Covid-19 Diagnostics Initiative (VCDI), Andrea Pauli, Julius Brennecke
bioRxiv 2020.06.23.166397; doi: https://doi.org/10.1101/2020.06.23.166397

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