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Updated: Jun 10, 2026
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A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons
Published on: November 14, 2018
Digital Hydrogel Fluorescent-Enhancing Microspheres via CRISPR/Cas12a for Amplification-Free Relative Quantification
Xueer Yin1,2, Qiangyuan Xiong1,2, Taowei Shu1
1Anhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China.
Analytical Chemistry
|June 8, 2026
Summary
This study introduces CrisprDEM, a novel CRISPR/Cas12a diagnostic system using hydrogel microspheres for ultrasensitive, amplification-free nucleic acid detection. It achieves high sensitivity and specificity for respiratory adenovirus detection, outperforming traditional methods.
Area of Science:
- Molecular Diagnostics
- Biotechnology
- Biosensing
Background:
- CRISPR/Cas systems offer diagnostic potential but struggle with weak signals and quantification in amplification-free assays.
- Existing methods lack the sensitivity and specificity required for early disease detection.
Purpose of the Study:
- To develop an ultrasensitive and quantitative amplification-free nucleic acid detection system using CRISPR/Cas12a.
- To enhance signal amplification and spatial resolution for improved diagnostic performance.
Main Methods:
- Developed CrisprDEM, a CRISPR/Cas12a-based digital hydrogel fluorescent-enhancing microsphere system.
- Integrated hydrogel microsphere (HM) confinement, microfluidic digital imaging, and machine learning.
- Utilized Intelligent Bead Analysis Software for quantification via positive bead ratio (PBR).
Main Results:
- Achieved 1000-fold signal amplification compared to homogeneous methods using HMs.
- Demonstrated a detection sensitivity of 10 aM for respiratory adenovirus with high specificity.
- Clinical sample validation showed consistency with real-time quantitative polymerase chain reaction (qPCR) and good PBR-Ct value linearity.
Conclusions:
- CrisprDEM provides ultrasensitive and quantitative amplification-free nucleic acid detection.
- The system shows promise for point-of-care diagnostics and early infection detection.
- Expands the capabilities of CRISPR-based diagnostic tools.
