Related Experiment Video
Updated: Aug 23, 2025
Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
Published on: August 16, 2024
Microfluidic space coding for multiplexed nucleic acid detection via CRISPR-Cas12a and recombinase polymerase
Zhichen Xu1,2, Dongjuan Chen3, Tao Li1,2
1State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, National Centre for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology-Wuhan National Laboratory for Optoelectronics, Chinese Academy of Sciences, Wuhan, 430071, China.
A new nucleic acid testing platform, MiCaR, enables fast, inexpensive, and multiplexed detection of up to 30 targets using microfluidics and CRISPR-Cas12a. This versatile tool accurately identifies HPV and respiratory viruses in patient samples.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Genomics
Background:
- Multiplexed nucleic acid detection is crucial for human health but faces challenges in speed, cost, and scalability.
- Existing methods often require multiple probes or complex procedures, limiting their widespread application.
Purpose of the Study:
- To develop an efficient, cost-effective, and highly multiplexed nucleic acid testing platform.
- To demonstrate the platform's capability in detecting clinically relevant pathogens, including human papillomavirus (HPV) and respiratory viruses.
Main Methods:
- Integration of a microfluidic device with CRISPR-Cas12a and multiplex recombinase polymerase amplification (RPA).
- Utilization of microfluidic space coding for simultaneous detection of multiple targets with a single fluorescence probe.
- Development of a rapid assay with a total run time of 40 minutes.
Main Results:
- Achieved a low limit of detection of 0.26 attomole.
- Successfully detected up to 30 nucleic acid targets simultaneously.
- Demonstrated high sensitivity (97.8%) and specificity (98.1%) for nine HPV subtypes in 100 patient samples.
- Validated the platform's generalizability by detecting eight common respiratory viruses.
Conclusions:
- The MiCaR platform offers a powerful solution for rapid, sensitive, and multiplexed nucleic acid detection.
- This versatile technology holds significant potential for clinical diagnostics and infectious disease surveillance.
- MiCaR is poised for broad adoption in various multiplexed nucleic acid testing applications.
