A Droplet-Free and Oil-Free Microfluidics Based on Self-Pumping Hydrogel for Point-of-Care Multiplex Digital Nucleic...

Area of Science:

• Biomaterials Engineering
• Molecular Diagnostics
• Microfluidics

Background:

• Digital PCR (dPCR) is crucial for nucleic acid quantification in diagnostics and safety.
• Current dPCR platforms face limitations in field applicability and multiplexing due to complex fluidics.

Purpose of the Study:

• To develop a self-pumping hydrogel chip for multiplex digital nucleic acid detection.
• To enable simple, low-cost, and portable point-of-care dPCR applications.

Main Methods:

• Utilized hydrogel's water-absorbing properties for autonomous fluid transport.
• Integrated hydrogel network for digital amplification, eliminating external pumps.
• Developed a multiplex assay with four detection zones for simultaneous pathogen quantification.

Main Results:

• Achieved fully passive operation with high sensitivity and single-copy quantitative accuracy.
• Demonstrated excellent storage stability and inhibitor tolerance of the hydrogel chip.
• Successfully performed simultaneous on-site digital quantification of three bacterial pathogens within 35 minutes.

Conclusions:

• The self-pumping hydrogel chip provides a simple, scalable platform for advanced digital nucleic acid analysis.
• This technology facilitates portable and high-throughput point-of-care testing.
• Overcomes limitations of traditional dPCR for in-field applications.