Digital-to-Droplet Microfluidic Platform Powered Digital Isothermal Nucleic Acid Amplification for On-Demand...

Area of Science:

• Biotechnology
• Microfluidics
• Molecular Diagnostics

Background:

• Severe infectious disease diagnosis faces challenges due to variable sample timing, diverse targets, and rapid turnaround needs.
• Current methods require development of on-demand detection strategies for timely etiological diagnosis.

Purpose of the Study:

• To introduce an automated digital isothermal nucleic acid amplification (dINA) assay using a digital-to-droplet (D2D) microfluidic platform.
• To enable on-demand detection methodologies for severe infectious diseases.

Main Methods:

• Developed a D2D microfluidic system for continuous droplet manipulation and interfacing with multiple units.
• Integrated digital microfluidics (DMF) for automated nucleic acid extraction and loop-mediated isothermal amplification (LAMP) reagent mixing.
• Utilized picoliter-scale droplets for dINA analysis after LAMP reaction in microliter droplets.

Main Results:

• Demonstrated on-demand processing of samples upon receipt, reducing turnaround times.
• Achieved substantial scalability in diagnostic throughput.
• Provided absolute quantification of nucleic acids without reliance on calibration standards.
• Successfully detected viruses causing post-transplantation infections in a proof-of-concept study.

Conclusions:

• The D2D platform facilitates efficient and adaptable on-demand detection of pathogens in severe infectious diseases.
• This automated system addresses limitations of traditional diagnostic approaches, improving timely diagnosis.