A lyophilized open-source RT-LAMP assay for molecular diagnostics in resource-limited settings
A critical bottleneck for equitable access to population-scale molecular diagnostics is the limited availability of rapid, inexpensive point-of-care tests, especially in low- and middle-income countries. Here, we developed an open-source reverse transcription loop-mediated isothermal amplification (RT-LAMP) molecular assay for pathogen detection. It is based on nonproprietary enzymes, namely, HIV-1 reverse transcriptase, <i>Bst</i> LF DNA polymerase, and UDG BMTU thermolabile uracil-DNA glycosylase. Formulated as liquid or lyophilized reaction mixtures, these reagents enable sensitive colorimetric detection of respiratory samples without the need for prior nucleic acid isolation. We evaluated our lyophilized RT-LAMP assay on clinical samples with suspected COVID-19 infection, demonstrating high sensitivity and 100% specificity compared with the gold-standard RT-qPCR. Reaction performance was unaffected by prolonged storage of lyophilized reagents at ambient or elevated temperatures. As a proof of concept, we evaluated the robustness and ease of use of lyophilized RT-LAMP reaction mixes through independent laboratory testing of COVID-19 samples in Ghana. Overall, our open-source RT-LAMP assay provides a flexible and scalable point-of-care test that can be adapted for rapid detection of various pathogens in resource-limited settings.
Authors
- Lucia L. Prieto-Godino (ORCID: https://orcid.org/0000-0002-2980-362X)
- Martin Mátl (ORCID: https://orcid.org/0000-0002-7215-3386)
- Thomas O. Auer (ORCID: https://orcid.org/0000-0003-3442-0567)
- Gordon A. Awandare (ORCID: https://orcid.org/0000-0002-8793-3641)
- Dominik Handler (ORCID: https://orcid.org/0000-0002-1059-4960)
- Julius Brennecke (ORCID: https://orcid.org/0000-0002-5141-0814)
- Irina Grishkovskaya (ORCID: https://orcid.org/0000-0003-0164-9373)
- Andrea Pauli (ORCID: https://orcid.org/0000-0001-9646-2303)
- Josef Penninger (ORCID: https://orcid.org/0000-0002-8194-3777)
- Luis Menéndez‐Arias (ORCID: https://orcid.org/0000-0002-1251-6640)
- Max J. Kellner (ORCID: https://orcid.org/0000-0002-5404-0628)
- Felix Ansah (ORCID: https://orcid.org/0000-0002-6928-2363)
- Benedikt Bauer (ORCID: https://orcid.org/0000-0002-9729-9568)
- Robert Heinen
Institutions
- Institute of Molecular Biotechnology (AT)
- University of Ghana (GH)
- University of Fribourg (CH)
- Research Institute of Molecular Pathology (AT)
- The Francis Crick Institute (GB)
- Helmholtz Centre for Infection Research (DE)
- Institute of Molecular Biology and Biotechnology (BG)
- Vienna Biocenter (AT)
- Centro de Biología Molecular Severo Ochoa (ES)
- Universidad Autónoma de Madrid (ES)
- Medical University of Vienna (AT)
Publication Details
- Journal
- Life Science Alliance
- Published
- 2025-07-14
- DOI
- https://doi.org/10.26508/lsa.202403167
- Citations
- 1
- Primary Topic
- Biosensors and Analytical Detection
- Type
- article
- Field-Weighted Citation Impact
- 0.67
Funders
- Fundación Ramón Areces
- Wellcome Trust
- European Federation of Pharmaceutical Industries and Associations
- Ministerio de Ciencia, Innovación y Universidades
- European Commission
- Volkswagen Foundation
- Vienna Science and Technology Fund
- Österreichischen Akademie der Wissenschaften
- Austrian Science Fund
- Universität Wien
- Vetenskapsrådet
- Österreichische Forschungsförderungsgesellschaft
- Medizinische Universität Wien
- Universität für Bodenkultur Wien
- T. Von Zastrow Foundation
- National Institutes of Health
- European Research Council
- Agencia Estatal de Investigación
- National Cancer Institute