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Published on: December 23, 2022
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Paper-based loop-mediated isothermal amplification and CRISPR integrated platform for on-site nucleic acid testing of
Anindita Sen1, Manaswini Masetty2, Sasanka Weerakoon2
1DNAiTECH Ltd, Marlborough Research Center, 2650 State Highway 1, Grovetown, Blenheim, Marlborough, 7202, New Zealand.
Biosensors & Bioelectronics
|April 23, 2024
Summary
We developed a paper-based diagnostic platform integrating Loop-mediated isothermal amplification (LAMP) and CRISPR technology for sensitive nucleic acid detection. This low-cost, smartphone-operated system offers rapid, on-site testing capabilities for various pathogens.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Point-of-Care Testing
Background:
- Traditional nucleic acid amplification methods often require complex equipment and thermal cycling.
- CRISPR-Cas12a systems offer high specificity for molecular detection, reducing false positives.
- Paper-based diagnostics are attractive for low-resource settings due to their simplicity and low cost.
Purpose of the Study:
- To develop and validate a novel paper-based platform for nucleic acid testing.
- To integrate Loop-mediated isothermal amplification (LAMP) with CRISPR technology for enhanced sensitivity and specificity.
- To create a user-friendly, smartphone-operated system for on-site molecular diagnostics.
Main Methods:
- Development of a paper-based platform (PLACID) integrating LAMP and CRISPR-Cas12a.
- Optimization of amplification and detection directly on paper.
- Engineering of a smartphone-controlled system with IR heating and fluorescence detection.
- Integration of equipment-free sample preparation technology.
Main Results:
- The PLACID platform demonstrated high specificity and sensitivity with a limit of detection of 50 copies/μL.
- Successful detection of SARS-CoV-2 RNA fragments in saliva, E. coli in soil, and pathogenic E. coli in fecal samples.
- The paper-based chips exhibited a shelf life of several weeks, suitable for field deployment.
Conclusions:
- The PLACID platform offers a simplified, sensitive, and specific approach to nucleic acid testing.
- The integrated system is suitable for on-site diagnostics, including infectious disease surveillance and environmental monitoring.
- This technology has the potential to improve access to molecular diagnostics in resource-limited settings.
