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Low-Cost Arduino Reverse Transcriptase Loop-Mediated Isothermal Amplification (RT-LAMP) for Sensitive Nucleic Acid Detection

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Publicado en:Biosensors vol. 14, no. 3 (2024), p. 128
Autor principal: Bruno Dias Camargo
Otros Autores: Stracke, Mateus Cassaboni, Heloisa Bruna Soligo Sanchuki, Viviane Klassen de Oliveira, Ancelmo, Hellen Cristina, Bordin, Dayanne Mozaner, Marchini, Fabricio Klerynton, Emilson Ribeiro Viana, Blanes, Lucas
Publicado:
MDPI AG
Materias:
Autodesk Inc
United States > US
Germany
Printed circuits
Software
Nucleic acids
Gene amplification
Aluminum
Severe acute respiratory syndrome coronavirus 2
Heating
Ribonucleic acid > RNA
Circuit boards
Biosensors
Heat
Colorimetry
Color
Low cost
Sensitivity
Cooling systems
Pandemics
Sensors
Organic light emitting diodes
Sensitivity analysis
Sensor arrays
RNA-directed DNA polymerase
Methods
Phenols
Multiplexer demultiplexers
Environmental
Acceso en línea:Citation/Abstract
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Resumen:This work presents a low-cost transcription loop-mediated isothermal amplification (RT-LAMP) instrument for nucleic acid detection, employing an Arduino Nano microcontroller. The cooling system includes customized printed circuit boards (PCBs) that serve as electrical resistors and incorporate fans. An aluminum block is designed to accommodate eight vials. The system also includes two PCB heaters—one for sample heating and the other for vial lid heating to prevent condensation. The color detection system comprises a TCS3200 color 8-sensor array coupled to one side of the aluminum heater body and a white 8-LED array coupled to the other side, controlled by two Multiplexer/Demultiplexer devices. LED light passes through the sample, reaching the color sensor and conveying color information crucial for detection. The top board is maintained at 110 ± 2 °C, while the bottom board is held at 65 ± 0.5 °C throughout the RT-LAMP assay. Validation tests successfully demonstrated the efficacy of the colorimetric RT-LAMP reactions using SARS-CoV-2 RNA amplification as a sample viability test, achieving 100% sensitivity and 97.3% specificity with 66 clinical samples. Our instrument offers a cost-effective (USD 100) solution with automated result interpretation and superior sensitivity compared to visual inspection. While the prototype was tested with SARS-CoV-2 RNA samples, its versatility extends to detecting other pathogens using alternative primers, showcasing its potential for broader applications in biosensing.
ISSN:2079-6374
DOI:10.3390/bios14030128
Fuente:Health & Medical Collection