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Updated: May 16, 2026
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MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
Published on: October 7, 2025
Simple yet sensitive MicroRNA detection using allosteric probe-initiated triple amplification and Cas13a/crRNA-based
1Wenzhou TCM Hospital of Zhejiang Chinese Medical University, Orthopedics and Traumatology, Wenzhou City, Zhejiang Province, 325000, China. gaogon0@163.com.
Summary
This study introduces a sensitive platform for detecting microRNAs (miRNAs) crucial for diagnosing degenerative disc disease. The novel method combines allosteric probes and CRISPR-Cas13a for highly accurate and rapid miRNA quantification.
Area of Science:
- Biotechnology
- Molecular Biology
- Diagnostics
Background:
- MicroRNAs (miRNAs) are vital biomarkers for degenerative disc disease (DDD).
- Clinical detection of miRNAs is challenging due to low abundance and sequence similarity.
- Sensitive and specific detection methods are needed for early DDD diagnosis.
Purpose of the Study:
- To develop a highly sensitive platform for miRNA detection.
- To overcome challenges in miRNA quantification for degenerative disc disease diagnosis.
- To enable early and minimally invasive diagnosis of DDD.
Main Methods:
- Integrated an allosteric probe-initiated triple-cycle amplification strategy with the CRISPR-Cas13a/crRNA system.
- Utilized polymerase extension, nicking enzyme recycling, and T7 RNA transcription for signal amplification.
- Employed Cas13a collateral cleavage activity on reporter RNAs for fluorescence signal generation.
Main Results:
- Achieved an ultra-low detection limit of 548 aM with a wide dynamic range (1 fM to 100 pM).
- Demonstrated excellent specificity, distinguishing target miRNA-155 from similar sequences.
- Showcased operational simplicity, rapid detection (60 min), and robustness in serum samples.
Conclusions:
- The developed platform offers a versatile and powerful tool for miRNA quantification.
- This method shows strong potential for clinical translation in early DDD diagnosis.
- The integrated approach advances miRNA detection for various pathologies.
