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Updated: Mar 15, 2026
MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
Published on: October 7, 2025
A simple G-quadruplex molecular beacon-based biosensor for highly selective detection of microRNA
Hui Zhou1, Chao Yang1, Huifang Chen2
1College of Life and Environmental Sciences, Gannan Normal University, Ganzhou 341000, PR China; Key Laboratory of Organo-Pharmaceutical Chemistry, College of Chemistry and Chemical Engineering, PR China; State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, PR China.
This study introduces a novel biosensor for detecting microRNAs (miRNAs) with high single-base precision. The G-quadruplex molecular beacon (G4MB) and duplex-specific nuclease (DSN) system significantly improves discrimination between similar miRNA sequences.
Area of Science:
- Biotechnology
- Molecular Biology
- Analytical Chemistry
Background:
- MicroRNA (miRNA) family members often differ by only a single nucleotide.
- High sequence homology among miRNAs presents a significant challenge for selective detection and analysis.
- Accurate miRNA profiling is crucial for understanding biological processes and disease mechanisms.
Purpose of the Study:
- To develop a highly selective and sensitive amplification biosensor for microRNA (miRNA) detection.
- To overcome the challenge of single-base variation in miRNA analysis.
- To create a robust platform for miRNA detection applicable to clinical diagnostics.
Main Methods:
- Development of a G-quadruplex molecular beacon (G4MB) as a recognition probe, utilizing a G4 motif stem.
- Integration of duplex-specific nuclease (DSN) for signal amplification through target miRNA recycling.
- Utilizing the protective G4 motif to prevent non-specific cleavage and reduce background fluorescence.
Main Results:
- The G4MB-DSN biosensor demonstrated high single-base selectivity in miRNA detection.
- The signal response difference between similar miRNA sequences with a one-base difference was reduced from 24% to 6% compared to traditional methods.
- The biosensor showed applicability in analyzing cancer cell samples and multiplexed detection of different miRNA targets.
Conclusions:
- The developed G4MB-DSN biosensor offers a simple, highly selective, and sensitive method for miRNA detection.
- This approach effectively distinguishes between miRNAs with single-base variations, overcoming a major analytical challenge.
- The biosensor's performance in real samples and multiplexing capability positions it as a promising tool for clinical diagnostics.
