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

Global efforts to combat the Covid-19 pandemic caused by SARS-CoV-2 still heavily rely 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 transc...

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Published inbioRxiv
Main Authors Kellner, Max J, Ross, James J, Schnabl, Jakob, Dekens, Marcus Ps, Heinen, Robert, Grishkovskaya, Irina, Bauer, Benedikt, Stadlmann, Johannes, Menendez-Arias, Luis, Fritsche-Polanz, Robert, Traugott, Marianna, Seitz, Tamara, Zoufaly, Alexander, Foedinger, Manuela, Wenisch, Christoph, Zuber, Johannes, Vienna Covid-19 Diagnostics Initiative (Vcdi), Pauli, Andrea, Brennecke, Julius
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 13.09.2021
Subjects
Colorimetry
Coronaviruses
COVID-19
Gene amplification
Lysis
Pandemics
Severe acute respiratory syndrome coronavirus 2
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Summary:Global efforts to combat the Covid-19 pandemic caused by SARS-CoV-2 still heavily rely 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. We present a rapid, robust, 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 open source enzymes that can 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. Footnotes * Updated the affiliation and email address of the first and co-corresponding author Max Kellner; Updated the abstract and introduction to reflect that vaccines are now available but not yet for everyone in the world.
DOI:10.1101/2020.06.23.166397