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

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Isothermal Amplification Technologies

Background:

• Nucleic acid detection is crucial for diagnosing infectious diseases.
• Existing methods can be expensive and require complex equipment.
• Loop-mediated isothermal amplification (LAMP) offers a promising alternative for rapid, point-of-care diagnostics.

Purpose of the Study:

• To develop and validate a low-cost, automated instrument for colorimetric RT-LAMP assays.
• To assess the instrument's performance in detecting SARS-CoV-2 RNA.
• To provide a cost-effective solution for nucleic acid detection.

Main Methods:

• An Arduino Nano microcontroller controlled a custom-designed instrument with PCB heaters and fans.
• A colorimetric detection system utilized a TCS3200 sensor array and LED array for real-time analysis.
• The instrument maintained precise temperatures for the RT-LAMP assay (top board at 110°C, bottom board at 65°C).

Main Results:

• The instrument successfully performed colorimetric RT-LAMP reactions for SARS-CoV-2 RNA detection.
• Validation tests with 66 clinical samples achieved 100% sensitivity and 97.3% specificity.
• The system provided automated result interpretation with superior sensitivity compared to visual inspection.

Conclusions:

• The developed low-cost RT-LAMP instrument is a viable and effective tool for nucleic acid detection.
• Its affordability and high performance make it suitable for broader applications in biosensing and infectious disease diagnostics.
• The instrument's design demonstrates potential for detecting various pathogens with appropriate primer modifications.