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Point-of-care CRISPR-based Diagnostics with Premixed and Freeze-dried Reagents
Published on: August 16, 2024
An integrated centrifugal microfluidic CRISPR-based diagnostics platform for multiplexed point-of-care testing of
Lingyun Li1, Hongjuan Wei1, Muyun Wei2
1Bioinformatics Center of AMMS, Beijing, 100850, PR China.
Biosensors & Bioelectronics
|August 3, 2026
Summary
The iCARD platform offers rapid, multiplexed point-of-care testing for respiratory pathogens. This integrated centrifugal assay system provides accurate results in under 40 minutes, improving clinical triage and infection control.
Area of Science:
- Molecular Diagnostics
- Microfluidics
- CRISPR Technology
Background:
- Accurate, rapid, multiplexed point-of-care testing (POCT) for respiratory pathogens is crucial for effective clinical triage and infection control.
- Existing POCT platforms often compromise on sensitivity, throughput, or operational simplicity.
- There is a need for advanced diagnostic tools that overcome these limitations in decentralized settings.
Purpose of the Study:
- To develop and validate the iCARD (integrated Centrifugal Assay on a Rotating Disc) platform, a novel microfluidic molecular diagnostic system.
- To enable rapid, automated, and multiplexed detection of respiratory pathogens at the point of care.
- To assess the diagnostic accuracy and performance of the iCARD platform using clinical samples.
Main Methods:
- Development of the iCARD platform, integrating centrifugal microfluidics with single-step CRISPR/Cas13a kinetics.
- Pre-loading of lyophilized reagents onto a polymethyl methacrylate disc for automated sample processing.
- Engineering a novel resin-based nucleic acid purification and concentration method for throat swabs.
- Utilizing a portable fluorescence analyzer for automated, multiplexed pathogen detection.
Main Results:
- The iCARD system achieved parallel screening of six respiratory pathogens from four samples within 40 minutes.
- Analytical validation demonstrated single-copy sensitivity (limit of detection: 0.25-1.0 copies/μL) with no cross-reactivity.
- Clinical validation on 94 patient throat swabs showed high diagnostic accuracy: 95.5% sensitivity and 100.0% specificity.
Conclusions:
- The iCARD platform is a robust, high-throughput, and accurate diagnostic tool for decentralized molecular screening.
- This system addresses the limitations of existing POCT platforms by offering streamlined operation and high performance.
- The iCARD platform holds significant potential for improving epidemiological surveillance and infection control strategies.
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