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bioRxiv · 10.1101/2020.06.23.166397

Scalable, rapid and highly sensitive isothermal detection of SARS-CoV-2 for laboratory and home testing

Kellner, M. J.·Ross, J. J.·Schnabl, J.·Dekens, M. P. S.·Heinen, R.·Tanner, N. A.·Fritsche-Polanz, R.·Traugott, M.·Seitz, T.·Zoufaly, A.·Foedinger, M.·Wenisch, C.·Zuber, J.·Vienna Covid-19 Diagnostics Initiative (VCDI),·Pauli, A.·Brennecke, J.
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Abstract

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.

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Kellner, M. J., Ross, J. J., Schnabl, J., Dekens, M. P. S., Heinen, R., Tanner, N. A., Fritsche-Polanz, R., Traugott, M., Seitz, T., Zoufaly, A., Foedinger, M., Wenisch, C., Zuber, J., Vienna Covid-19 Diagnostics Initiative (VCDI),, Pauli, A., Brennecke, J.. 2020-06-23. Scalable, rapid and highly sensitive isothermal detection of SARS-CoV-2 for laboratory and home testing. https://doi.org/10.1101/2020.06.23.166397

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