Low-Cost, Open-Source Device for High-Performance Fluorescence Detection of Isothermal Nucleic Acid Amplification Reactions
Open Access
- 14 September 2021
- journal article
- research article
- Published by American Chemical Society (ACS) in ACS Biomaterials Science & Engineering
- Vol. 7 (10), 4982-4990
- https://doi.org/10.1021/acsbiomaterials.1c01105
Abstract
The ability to detect SARS-CoV-2 is critical to implementing evidence-based strategies to address the COVID-19 global pandemic. Expanding SARS-CoV-2 diagnostic ability beyond well-equipped laboratories widens the opportunity for surveillance and control efforts. However, such advances are predicated on the availability of rapid, scalable, accessible, yet high-performance diagnostic platforms. Methods to detect viral RNA using reverse transcription loop-mediated isothermal amplification (RT-LAMP) show promise as rapid and field-deployable tests; however, the per-unit costs of the required diagnostic hardware can be a barrier for scaled deployment. Here, we describe a diagnostic hardware configuration for LAMP technology, named the FABL-8, that can be built for approximately US$380 per machine and provide results in under 30 min. Benchmarking showed that FABL-8 has a similar performance to a high-end commercial instrument for detecting fluorescence-based LAMP reactions. Performance testing of the instrument with RNA extracted from a SARS-CoV-2 virus dilution series revealed an analytical detection sensitivity of 50 virus copies per microliter—a detection threshold suitable to detect patient viral load in the first few days following symptom onset. In addition to the detection of SARS-CoV-2, we show that the system can be used to detect the presence of two bacterial pathogens, demonstrating the versatility of the platform for the detection of other pathogens. This cost-effective and scalable hardware alternative allows democratization of the instrumentation required for high-performance molecular diagnostics, such that it could be available to laboratories anywhere—supporting infectious diseases surveillance and research activities in resource-limited settings.Keywords
Funding Information
- Australian government Medical Research Future Fund (MRF2002317)
This publication has 33 references indexed in Scilit:
- Detection of dengue viruses using reverse transcription-loop-mediated isothermal amplificationBMC Infectious Diseases, 2013
- Rapid and Sensitive Detection of Mycobacterium ulcerans by Use of a Loop-Mediated Isothermal Amplification TestJournal of Clinical Microbiology, 2012
- LAMP-based method for a rapid identification of Legionella spp. and Legionella pneumophilaApplied Microbiology and Biotechnology, 2011
- Development of LAMP and Real-Time PCR Methods for the Rapid Detection of Xylella fastidiosa for Quarantine and Field ApplicationsPhytopathology®, 2010
- LAMP Using a Disposable Pocket Warmer for Anthrax Detection, a Highly Mobile and Reliable Method for Anti-BioterrorismJapanese Journal of Infectious Diseases, 2010
- Potential of LAMP for detection of plant pathogens.CABI Reviews, 2008
- Rapid Detection and Quantification of Japanese Encephalitis Virus by Real-Time Reverse Transcription Loop-Mediated Isothermal AmplificationMicrobiology and Immunology, 2006
- Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virusArchiv für die gesamte Virusforschung, 2006
- Loop-Mediated Isothermal Amplification for Direct Detection of Mycobacterium tuberculosis Complex, M. avium , and M. intracellulare in Sputum SamplesJournal of Clinical Microbiology, 2003
- Loop-mediated isothermal amplification of DNANucleic Acids Research, 2000