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

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 a...

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Detalles Bibliográficos
Autores principales: Kellner, Max J., Ross, James J., Schnabl, Jakob, Dekens, Marcus P. S., Matl, Martin, Heinen, Robert, Grishkovskaya, Irina, Bauer, Benedikt, Stadlmann, Johannes, Menéndez-Arias, Luis, Straw, Andrew D., Fritsche-Polanz, Robert, Traugott, Marianna, Seitz, Tamara, Zoufaly, Alexander, Födinger, Manuela, Wenisch, Christoph, Zuber, Johannes, Pauli, Andrea, Brennecke, Julius
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9011764/
https://www.ncbi.nlm.nih.gov/pubmed/35433827
http://dx.doi.org/10.3389/fmolb.2022.801309
Descripción
Sumario: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.