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

• DOI: 10.3389/fmolb.2022.801309
• Publication Date: 2022-04-01T11:17:49Z
• Authors: Max J. Kellner; James J. Ross; Jakob Schnabl; Marcus P. S. Dekens; Martin Matl; Robert Heinen; Irina Grishkovskaya; Benedikt Bauer; Johannes Stadlmann; Luis Menéndez-Arias; Andrew D. Straw; Robert Fritsche-P...; Marianna Traugott; Tamara Seitz; Alexander Zoufaly; Manuela Födinger; Christoph Wenisch; Johannes Zuber; Vienna COVID-19 ...; Andrea Pauli; Julius Brennecke
• Keywords: Open-source; PURIFICATION; isothermal amplification; QH301-705.5; COVID-19 diagnostics; SARS-CoV-2; 106002 Biochemie; DNA-POLYMERASE; coronavirus; 106002 Biochemistry; FIDELITY; 3. Good health; Coronavirus; Isothermal amplification; REVERSE-TRANSCRIPTASE; LAMP; MEDIATED ISOTHERMAL AMPLIFICATION; Molecular Biosciences; 106052 Zellbiologie; Biology (General); 106052 Cell biology; open-source; RT-LAMP

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.