Low-Cost, Open-Source Device for High-Performance Fluorescence Detection of Isothermal Nucleic Acid Amplification Reactions
| Autor: | Benjamin P Howden, Ian R. Monk, Timothy P. Stinear, Jean Y. H. Lee, Koen Vandelannoote, Andrew H. Buultjens, Liam K. R. Sharkey |
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| Rok vydání: | 2021 |
| Předmět: |
Rapid diagnostic test
Computer science business.industry SARS-CoV-2 Point-of-care testing Biomedical Engineering Loop-mediated isothermal amplification COVID-19 Nucleic acid amplification technique Molecular diagnostics Biomaterials Molecular Diagnostic Techniques Embedded system Scalability Humans RNA Viral Instrumentation (computer programming) business Viral load Nucleic Acid Amplification Techniques |
| Zdroj: | ACS biomaterials scienceengineering. 7(10) |
| ISSN: | 2373-9878 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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