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iso-μmGene: an isothermal amplification-based portable microfluidic system for simple, reliable and flexibly
Runtao Zhong1, Shilin Liu, Guohao Zhang
1Institute of Environmental Systems Biology, Dalian Maritime University, 1 Linghai Road, Dalian 116026, China. rtzhong@dlmu.edu.cn yqsun@dlmu.edu.cn.
The Analyst
|May 28, 2020
Summary
A portable microfluidic system (iso-μmGene) enables rapid genetic identification and quantification of microbial pathogens. This point-of-care device offers reliable, multiplexed detection for early disease diagnosis and treatment.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Microfluidics
Background:
- Accurate microbial pathogen identification is crucial for timely disease diagnosis and treatment.
- Existing methods often lack the speed, portability, or multiplexing capability required for point-of-care applications.
Purpose of the Study:
- To develop a simple, reliable, and flexibly multiplexed isothermal amplification-based portable microfluidic system (iso-μmGene) for microbial pathogen genetic testing.
- To enable on-demand analysis of multiple target genes from various samples with high throughput and portability.
Main Methods:
- Utilized disposable microfluidic chips with 10 or 18 wells and a device prototype with four chip holders.
- Integrated dehydrated primers, a single pipetting step, and a dedicated plastic shell for reliable loop-mediated isothermal amplification (LAMP) reaction sealing.
- Employed a thermoelectric cooler (TEC)-based temperature control module and a CCD-based fluorescence imaging module with a linear translation stage for real-time detection.
Main Results:
- Demonstrated parallel detection of microbial pathogens with a dynamic range of 2-2000 colony forming units (CFU) per reaction well.
- Achieved good intra- and inter-chip reproducibility for the genetic identification and quantification of Edwardsiella tarda and Vibrio harveyi.
- Validated the system's performance using crude lysates of aquaculture pathogens.
Conclusions:
- The iso-μmGene system offers a cost-effective, portable, and high-performance solution for point-of-care genetic testing.
- The device combines sophisticated functionality with the portability and affordability required for widespread diagnostic applications.
- This technology addresses the urgent need for rapid and reliable microbial pathogen detection in clinical and field settings.
