A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing
2022
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
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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