Frontiers in Molecular Biosciences (Apr 2022)

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

  • Max J. Kellner,
  • Max J. Kellner,
  • Max J. Kellner,
  • Max J. Kellner,
  • James J. Ross,
  • James J. Ross,
  • Jakob Schnabl,
  • Jakob Schnabl,
  • Marcus P. S. Dekens,
  • Martin Matl,
  • Martin Matl,
  • Robert Heinen,
  • Robert Heinen,
  • Irina Grishkovskaya,
  • Benedikt Bauer,
  • Johannes Stadlmann,
  • Johannes Stadlmann,
  • Johannes Stadlmann,
  • Luis Menéndez-Arias,
  • Andrew D. Straw,
  • Robert Fritsche-Polanz,
  • Marianna Traugott,
  • Tamara Seitz,
  • Alexander Zoufaly,
  • Manuela Födinger,
  • Manuela Födinger,
  • Christoph Wenisch,
  • Johannes Zuber,
  • Johannes Zuber,
  • Vienna COVID-19 Detection Initiative (VCDI),
  • Andrea Pauli,
  • Julius Brennecke

DOI
https://doi.org/10.3389/fmolb.2022.801309
Journal volume & issue
Vol. 9

Abstract

Read online

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.

Keywords