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CoID-LAMP: Color-Encoded, Intelligent Digital LAMP for Multiplex Nucleic Acid Quantification.
Kai Wu1,2, Qi Fang1,2, Zhantao Zhao1,2
1Department of Biomedical Engineering, Medical School, Shenzhen University, Shenzhen 518060, China.
Analytical Chemistry
|March 9, 2023
Summary
Color-encoded, intelligent digital loop-mediated isothermal amplification (CoID-LAMP) enables multiplex nucleic acid detection. This novel method uses droplet microfluidics and deep learning for accurate quantification, overcoming limitations of current assays.
Area of Science:
- Biomedical applications
- Molecular diagnostics
- Nucleic acid testing
Background:
- Multiplex digital nucleic acid tests are crucial in biomedicine.
- Existing fluorescent probe methods are difficult to optimize and limit applications.
Purpose of the Study:
- To develop a novel method for co-identification of multiple nucleic acid targets.
- To overcome limitations of current multiplex nucleic acid detection methods.
Main Methods:
- Developed color-encoded, intelligent digital loop-mediated isothermal amplification (CoID-LAMP).
- Utilized droplet microfluidics to pair primer and sample droplets.
- Employed deep learning for image analysis and droplet detection.
- Validated using fluorescent and brightfield dyes for multiplex assays.
Main Results:
- Established a reliable image analysis pipeline using deep learning.
- Demonstrated an 8-plex assay using fluorescent dyes with reliable coding and quantification.
- Showcased a 4-plex assay using brightfield dyes, indicating simplified optical requirements.
- Confirmed the capability for accurate multiplex nucleic acid quantification.
Conclusions:
- CoID-LAMP offers a versatile tool for multiplex nucleic acid quantification.
- The combination of droplet microfluidics and deep learning enhances assay performance.
- CoID-LAMP provides a promising alternative to existing digital nucleic acid testing methods.
