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Updated: Jun 26, 2026
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High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR
Published on: November 28, 2016
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Amplification-Free CRISPR-Cas System Integrated Centrifugal Digital Microfluidic Platform Developed for Multiplexed
Jing Zhang1, Longjie Li1, Yidan Zhu1
1Jiangsu Key Laboratory of Advanced Medical Analysis and Public Health, Nantong Key Laboratory of Public Health and Medical Analysis, School of Public Health, Nantong University, Nantong, Jiangsu 226019, P. R. China.
Analytical Chemistry
|November 16, 2025
Summary
This study developed a CRISPR-Cas9/Cas13a digital microfluidic chip for rapid, sensitive detection of respiratory pathogens like MRSA and H1N1. The platform offers high accuracy without preamplification, ideal for resource-limited settings.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Bioengineering
Background:
- Traditional PCR requires specialized equipment and personnel.
- Existing isothermal amplification methods can yield false positives.
- Sensitive CRISPR detection often necessitates nucleic acid preamplification.
Purpose of the Study:
- To develop an integrated CRISPR-Cas9/Cas13a detection platform on a centrifugal digital microfluidic chip.
- To overcome limitations of current diagnostic technologies for respiratory infectious diseases.
- To enable rapid, sensitive, and accurate multiplex pathogen detection.
Main Methods:
- Development and optimization of an off-chip CRISPR-Cas9/Cas13a dual nucleic acid detection system.
- Design and screening of an optimal centrifugal digital microfluidic chip structure.
- Integration of the optimized CRISPR system into the digital chip for pathogen detection.
Main Results:
- Achieved specific detection of MRSA DNA (173 pM) and H1N1 RNA (117 pM) off-chip.
- Optimized chip achieved a 99.6% droplet filling rate.
- On-chip detection reached subcopy sensitivity (0.7 copies/μL for MRSA, 1.2 copies/μL for H1N1) within 20 minutes.
- Demonstrated 100% accuracy for both positive and negative detection in simulated clinical samples.
Conclusions:
- The integrated platform enables sensitive, rapid, and accurate multiplex detection of respiratory pathogens without preamplification.
- This convenient, cost-effective, and contamination-resistant method is suitable for resource-constrained scenarios.
- The technology offers a reliable solution for infectious disease prevention and control.
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