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A disposable, integrated loop-mediated isothermal amplification cassette with thermally actuated valves
Changchun Liu1, Michael G Mauk1, Haim H Bau1
1Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, 229 Towne Building, 220 S. 33rd St., Philadelphia, PA 19104-6315 USA.
Microfluidics and Nanofluidics
|March 28, 2020
Summary
A new, low-cost, disposable polymer cassette enables rapid nucleic acid amplification using loop-mediated isothermal amplification (LAMP). This integrated system, featuring novel microvalves, offers sensitive detection for point-of-care applications.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Materials Science
Background:
- Loop-mediated isothermal amplification (LAMP) is a powerful nucleic acid amplification technique.
- Existing LAMP platforms often require complex infrastructure and trained personnel.
- There is a need for portable, cost-effective diagnostic tools for resource-limited settings.
Purpose of the Study:
- To design and fabricate an inexpensive, disposable, integrated polymer cassette for LAMP.
- To develop and characterize thermally actuated microvalves for the cassette.
- To demonstrate the utility of the cassette for rapid nucleic acid detection.
Main Methods:
- Fabrication of a polymer-based cassette with an integrated LAMP chamber.
- Development of single-use, thermally actuated valves using a PDMS-microsphere composite.
- Evaluation of valve performance under pressure.
- Actuation of the chamber and valves using thin-film heaters.
- Demonstration of LAMP and RT-LAMP for *Escherichia coli* DNA and RNA targets.
- Real-time detection of amplicons using a portable detector.
Main Results:
- The developed microvalves maintained integrity up to 200 kPa in the closed state.
- The integrated cassette successfully performed LAMP and RT-LAMP.
- The system achieved sensitive detection of as few as 10 target molecules per sample.
- Assay completion was achieved in under 1 hour.
- The system demonstrated high efficiency and reliability.
Conclusions:
- An inexpensive, disposable, integrated polymer cassette for LAMP has been successfully developed.
- The cassette system, incorporating novel microvalves, is suitable for rapid nucleic acid detection.
- This technology holds significant promise for point-of-care diagnostics and applications in resource-poor settings.
