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Updated: Mar 2, 2026
MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
Published on: October 7, 2025
Sensitive Quantification of MicroRNAs by Isothermal Helicase-Dependent Amplification
1College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Shandong Provincial Key Laboratory of Clean Production of Fine Chemicals, Shandong Normal University , Jinan 250014, China.
We developed a simple, rapid, and sensitive fluorescent assay for quantifying low-abundance microRNAs (messenger RNAs). This method uses isothermal helicase-dependent amplification for highly sensitive detection, aiding clinical diagnosis and biomedical research.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- MicroRNA (messenger RNA) dysregulation is linked to various diseases.
- Accurate microRNA quantification is crucial for clinical diagnosis and treatment.
- Existing methods lack sensitivity for low-abundance microRNAs or involve complex procedures.
Purpose of the Study:
- To develop a simple, rapid, and sensitive fluorescent method for label-free microRNA detection.
- To overcome the limitations of conventional quantification techniques for low-abundance microRNAs.
Main Methods:
- Utilized isothermal helicase-dependent amplification (HDA) for nucleic acid amplification.
- Designed a linear probe that protects against exonuclease I digestion upon hybridization with target microRNA.
- Employed a fluorescent readout for signal generation within 30 minutes.
Main Results:
- Achieved a low detection limit of 12.8 fM for microRNAs.
- Demonstrated a broad dynamic range from 100 fM to 10 nM.
- Successfully discriminated between different microRNA family members and quantified endogenous microRNAs in cancer cell samples.
Conclusions:
- The developed HDA-based fluorescent assay offers a sensitive, rapid, and simple approach for microRNA quantification.
- This method is suitable for detecting low-abundance microRNAs and analyzing endogenous microRNAs in biological samples.
- Presents a valuable tool for advancing biomedical research and clinical diagnostics.
