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Pipette-Tip-Enabled Digital Nucleic Acid Analyzer for COVID-19 Testing with Isothermal Amplification
Wenyao Zhang1, Kaixin Zheng1, Yang Ye1,2,3
1National Engineering Research Center for Optical Instruments, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310052, China.
Analytical Chemistry
|November 4, 2021
Summary
A new pipette-tip digital nucleic acid analyzer enables sensitive COVID-19 testing using digital loop-mediated isothermal amplification (dLAMP). This accessible method offers improved detection limits for rapid and reliable screening.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Nucleic Acid Analysis
Background:
- Accurate and accessible COVID-19 testing is crucial for public health.
- Digital nucleic acid amplification offers high sensitivity but often requires specialized equipment.
- Loop-mediated isothermal amplification (LAMP) provides a thermal cycler-free amplification method.
Purpose of the Study:
- To develop a low-cost, pipette-tip-based digital nucleic acid analyzer for COVID-19 detection.
- To demonstrate the feasibility of digital loop-mediated isothermal amplification (dLAMP) using common lab materials.
- To achieve high sensitivity and accessibility in nucleic acid testing.
Main Methods:
- Integration of a glass capillary into standard pipette tips for microdroplet generation via benchtop centrifugation.
- Utilizing digital loop-mediated isothermal amplification (dLAMP) to amplify the ORF1a/b gene, a target for COVID-19 screening.
- Performing Poisson analysis on fluorescent microdroplets for accurate quantification.
Main Results:
- Successfully generated monodisperse water-in-oil microdroplets within pipette tips.
- Achieved amplification of the target COVID-19 gene without a thermal cycler.
- Demonstrated a limit of detection one order of magnitude better than non-digital techniques, comparable to commercial dLAMP platforms.
Conclusions:
- A pipette-tip-enabled dLAMP system provides a sensitive and accessible platform for COVID-19 testing.
- This approach simplifies digital nucleic acid analysis, reducing reliance on expensive instrumentation.
- The developed method holds potential for widespread application in nucleic acid detection, particularly in resource-limited settings.
