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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.
  • Kellner MJ; Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Vienna, Austria.
  • Ross JJ; Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Vienna, Austria.
  • Schnabl J; Vienna BioCenter PhD Program, Doctoral School of the University of Vienna and Medical University of Vienna, Vienna, Austria.
  • Dekens MPS; MRC Laboratory of Molecular Biology, Cambridge, United Kingdom.
  • Matl M; Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Vienna, Austria.
  • Heinen R; Vienna BioCenter PhD Program, Doctoral School of the University of Vienna and Medical University of Vienna, Vienna, Austria.
  • Grishkovskaya I; Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Vienna, Austria.
  • Bauer B; Vienna BioCenter PhD Program, Doctoral School of the University of Vienna and Medical University of Vienna, Vienna, Austria.
  • Stadlmann J; Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Vienna, Austria.
  • Menéndez-Arias L; Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Vienna, Austria.
  • Straw AD; Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Vienna, Austria.
  • Fritsche-Polanz R; Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Vienna, Austria.
  • Traugott M; Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Vienna, Austria.
  • Seitz T; Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Vienna, Austria.
  • Zoufaly A; Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Vienna, Austria.
  • Födinger M; Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Vienna, Austria.
  • Wenisch C; Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Vienna, Austria.
  • Zuber J; Department of Chemistry, University of Natural Resources and Life Sciences, Vienna, Austria.
  • Pauli A; Centro de Biología Molecular "Severo Ochoa" (Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid), Madrid, Spain.
  • Brennecke J; Institute of Biology I and Bernstein Center Freiburg, Faculty of Biology, Albert-Ludwigs-University Freiburg, Freiburg, Germany.
Front Mol Biosci ; 9: 801309, 2022.
Article in English | MEDLINE | ID: covidwho-1793001
ABSTRACT
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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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study Language: English Journal: Front Mol Biosci Year: 2022 Document Type: Article Affiliation country: Fmolb.2022.801309

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study Language: English Journal: Front Mol Biosci Year: 2022 Document Type: Article Affiliation country: Fmolb.2022.801309