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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
AND-logic amplification enables one-pot co-detection of extracellular vesicle miRNA and APE1
1Department of Gastrointestinal Surgery, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, 362000, China.
Analytical Biochemistry
|May 26, 2026
Summary
This study introduces a novel one-pot assay for detecting extracellular vesicles (EVs) using dual biomarkers, including tumor-associated microRNAs (miRNAs) and the endonuclease APE1. This advanced liquid biopsy method significantly improves cancer detection accuracy compared to single-marker tests.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Extracellular vesicles (EVs) in body fluids are valuable for liquid biopsies due to their molecular cargo.
- Current EV assays often rely on single markers, limiting diagnostic performance by redundancy.
- Integrating multiple markers across different classes is challenging for EV analysis.
Purpose of the Study:
- To develop a one-pot dual-biomarker detection platform for extracellular vesicles (EVs).
- To utilize an AND logic gate for synergistic detection of tumor-associated miRNAs and endonuclease APE1.
- To enhance diagnostic performance for cancer detection using liquid biopsy.
Main Methods:
- Developed a one-pot platform integrating strand displacement amplification (SDA) and endonuclease APE1 cleavage.
- Employed an AND logic gate to combine signals from miRNA and APE1.
- Utilized cascading signal amplification for sensitive detection.
Main Results:
- Achieved a limit of detection of 8 fM for miRNA and 0.008 U/mL for APE1.
- Demonstrated a high area under the ROC curve (AUC) of 0.957 for gastric cancer detection in plasma EVs.
- Outperformed single-marker RT-qPCR miRNA analysis (AUC=0.823) in clinical samples.
Conclusions:
- The developed logic-gated framework effectively integrates multi-class EV biomarkers.
- This platform shows significant potential for early cancer screening and disease monitoring.
- The synergistic approach enhances diagnostic discrimination in liquid biopsies.
