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TriD-LAMP: A pump-free microfluidic chip for duplex droplet digital loop-mediated isothermal amplification analysis
Cui Wu1, Linbo Liu2, Zunzhong Ye3
1School of Biosystems Engineering and Food Science, Zhejiang University, 866 Yuhangtang Road, Hangzhou, Zhejiang, 310058, PR China; John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, 02138, USA.
Analytica Chimica Acta
|October 25, 2022
Summary
A new pump-free microfluidic chip enables digital loop-mediated isothermal amplification (dLAMP) for sensitive DNA detection. This integrated system simplifies sample processing for multiple targets in diagnostics.
Area of Science:
- Biotechnology
- Microfluidics
- Molecular Diagnostics
Background:
- Digital nucleic acid amplification offers sensitive, absolute quantification.
- Digital loop-mediated isothermal amplification (dLAMP) is robust for field detection but requires pumps for sample digitalization.
- Existing droplet dLAMP systems often necessitate droplet transfer post-digitalization or amplification.
Purpose of the Study:
- To develop and assess a pump-free microfluidic chip for duplex droplet dLAMP (TriD-LAMP).
- To enable integrated sample digitalization, amplification, and detection without droplet transfer.
- To demonstrate simultaneous quantification of multiple nucleic acid targets.
Main Methods:
- A custom microfluidic chip utilizing step emulsification for droplet generation.
- Integrated droplet generation (64 nozzles, <5% variation) and storage zones.
- Pump-free operation with oil containing fluorosurfactant to prevent droplet fusion and evaporation.
Main Results:
- Achieved accurate quantification of E. coli DNA down to 19.8 copies/μL.
- Demonstrated simultaneous quantification of two targets (duplex detection).
- The chip successfully performed all dLAMP stages without droplet transfer.
Conclusions:
- The integrated, pump-free TriD-LAMP chip simplifies digital nucleic acid amplification.
- This technology holds promise for multiplexed detection in clinical diagnostics and point-of-care settings.
- Eliminates the need for auxiliary pumps and droplet transfer steps, enhancing usability.
