Three-Dimensional-Printed Instrument for Isothermal Nucleic Acid Amplification with Real-Time Colorimetric Imaging

Area of Science:

• Molecular Biology
• Biotechnology
• Analytical Chemistry

Background:

• Isothermal amplification methods, like loop-mediated isothermal amplification (LAMP), are valued for their simplicity.
• Colorimetry is a common, yet subjective, method for monitoring LAMP reactions.
• Objective detection is needed to overcome limitations in visual interpretation of color changes.

Purpose of the Study:

• To develop a novel, automated, and integrated analysis system for objective colorimetric detection of isothermal amplification.
• To create a cost-effective, high-throughput platform adaptable to various colorimetric dyes.
• To validate the system's performance against conventional methods.

Main Methods:

• Development of a one-step, automated analysis system featuring 3D-printed convective heating.
• Integration of programmed image capture with simultaneous heating for real-time monitoring.
• Utilization of LabVIEW software for automated image analysis and objective colorimetric assessment.
• Validation using messenger-RNA targets and mock forensic samples.

Main Results:

• The developed system provides objective colorimetric analysis of isothermal amplification.
• Performance was comparable to conventional thermal cyclers and smartphone-based image analysis.
• The system demonstrated exceptional throughput, analyzing up to 96 samples concurrently.
• The device is cost-effective and adaptable to multiple colorimetric dyes.

Conclusions:

• The novel automated system enhances the objectivity and efficiency of isothermal amplification monitoring.
• This integrated platform offers a cost-effective and high-throughput solution for molecular diagnostics and research.
• The technology has potential applications in various fields requiring precise nucleic acid detection.