Acryloyl Gelatin as Polymer Additive to Droplet Digital LAMP for DNA Quantification

Area of Science:

• Biotechnology
• Microfluidics
• Molecular Diagnostics

Background:

• Droplet digital nucleic acid quantification (NAQ) is crucial for life analysis.
• Microgel particles enhance droplet functionalities for tandem analysis in NAQ.
• Limited material options exist for microgel-based droplet digital NAQ.

Purpose of the Study:

• Introduce acryloyl gelatin (GelA) as a novel polymer for droplet digital NAQ.
• Evaluate GelA's suitability for droplet microfluidics and nucleic acid amplification.
• Demonstrate GelA's utility in a SARS-CoV-2 detection assay.

Main Methods:

• Incorporation of photopolymerizable, biocompatible, and biodegradable GelA into droplets.
• Assessment of GelA's viscosity and thermal stability for microfluidic handling.
• Optimization of droplet digital loop-mediated amplification (LAMP) with GelA for SARS-CoV-2 N gene detection.

Main Results:

• GelA showed minimal thermal viscosity variation, aiding microfluidic production.
• GelA did not significantly impact droplet digital LAMP efficiency.
• SARS-CoV-2 detection achieved within 25-250,000 copies/test in 30 min using GelA-enhanced droplet digital LAMP.
• GelA droplets transformed into stable microgel particles for controlled product release.

Conclusions:

• Acryloyl gelatin expands material choices for droplet digital NAQ.
• GelA enhances the versatility of droplet digital LAMP assays.
• This advancement offers new possibilities for nucleic acid detection and analysis.