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Updated: Mar 28, 2026
MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
Published on: October 7, 2025
Visual Detection of Multiplex MicroRNAs Using Cationic Conjugated Polymer Materials
Yuanyuan Zhou1, Jiangyan Zhang1, Likun Zhao1
1Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Ministry of Education, Key Laboratory of Analytical Science and Technology of Hebei Province, College of Chemistry and Environmental Science, Hebei University , Baoding 071002, Hebei, P. R. China.
A new method uses duplex-specific nuclease (DSN) amplification and cationic conjugated polymers (CCPs) for simple, visual detection of microRNAs (miRNAs). This approach enables naked-eye identification of specific miRNA targets through fluorescence changes.
Area of Science:
- Biochemistry
- Molecular Biology
- Materials Science
Background:
- MicroRNA (miRNA) detection is crucial for understanding gene regulation and disease diagnostics.
- Existing methods for multiplex miRNA detection often lack simplicity, visual output, or specificity.
- Development of novel sensing platforms is needed for efficient and accessible miRNA analysis.
Purpose of the Study:
- To develop a simple, visual, and specific method for simultaneous detection of multiplex microRNAs (miRNAs).
- To integrate duplex-specific nuclease (DSN)-induced amplification with cationic conjugated polymer (CCP) materials for enhanced miRNA sensing.
- To enable direct, naked-eye visualization of miRNA detection through fluorescence resonance energy transfer (FRET) signal changes.
Main Methods:
- Utilized DNA probes labeled with fluorescein dye (FAM) and complementary to target miRNAs.
- Employed duplex-specific nuclease (DSN) for amplification via cycled probe digestion upon target miRNA binding.
- Integrated cationic conjugated polymers (CCPs) to facilitate fluorescence resonance energy transfer (FRET) between CCPs and FAM-labeled probes.
Main Results:
- Demonstrated that target miRNA presence leads to DSN-mediated probe digestion, reducing FRET efficiency and causing a visual fluorescence change.
- Achieved specific detection of miRNAs by monitoring FRET signal changes from CCP to FAM.
- Showcased simultaneous detection of multiplex miRNAs by designing an energy transfer cascade using CCP and differentially labeled DNA probes (FAM and Cy3).
Conclusions:
- The developed DSN-amplification and CCP-based platform offers a simple, visual, and specific method for miRNA detection.
- The system allows for naked-eye visualization of miRNA detection by triggering FRET signal changes.
- This approach holds promise for simultaneous detection of multiple miRNAs, advancing diagnostic and research capabilities.
