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A lyophilized open-source RT-LAMP assay for molecular diagnostics in resource-limited settings
Martin Matl1,2, Max J Kellner1,2,3,4, Felix Ansah5
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Vienna, Austria.
Life Science Alliance
|July 14, 2025
Summary
An open-source molecular assay using reverse transcription loop-mediated isothermal amplification (RT-LAMP) offers rapid, inexpensive pathogen detection. This point-of-care test is ideal for resource-limited settings, demonstrating high accuracy for COVID-19 detection.
Area of Science:
- Molecular Diagnostics
- Isothermal Amplification
- Point-of-Care Testing
Background:
- Equitable access to molecular diagnostics is hindered by the lack of rapid, affordable point-of-care tests, particularly in low- and middle-income countries.
- Existing diagnostic methods often require specialized equipment and trained personnel, limiting their applicability in resource-constrained environments.
Purpose of the Study:
- To develop an open-source, low-cost, and adaptable molecular assay for pathogen detection suitable for point-of-care use.
- To create a stable, easy-to-use diagnostic reagent that bypasses the need for nucleic acid isolation and complex laboratory infrastructure.
Main Methods:
- Development of an open-source reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay utilizing nonproprietary enzymes.
- Formulation of reagents into liquid or lyophilized mixtures for sensitive colorimetric detection of respiratory pathogens.
- Evaluation of the lyophilized RT-LAMP assay on clinical COVID-19 samples against RT-qPCR, assessing sensitivity, specificity, and storage stability.
- Independent laboratory testing in Ghana to assess robustness and usability in a resource-limited setting.
Main Results:
- The developed RT-LAMP assay demonstrated high sensitivity and 100% specificity for COVID-19 detection compared to RT-qPCR.
- Lyophilized reagents maintained reaction performance after prolonged storage at various temperatures.
- Independent testing confirmed the assay's robustness and ease of use in a real-world, resource-limited setting.
Conclusions:
- The open-source RT-LAMP assay provides a flexible, scalable, and cost-effective solution for pathogen detection at the point of care.
- This assay has the potential to significantly improve diagnostic capabilities in low-resource settings, enabling rapid response to infectious disease outbreaks.
- The adaptable nature of the assay allows for its application in detecting a wide range of pathogens beyond COVID-19.
