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A Novel Dual Signal Amplification Strategy Integrating APE1-Assisted DNA Walker with Rolling Circle Amplification for
Huihui Yang1, Ziyue Qiu2, Shishi Huang2
1Department of Pharmacy, The Eighth Affiliated Hospital, Sun Yat-Sen University, Shenzhen 518033, P. R. China.
Analytical Chemistry
|March 6, 2026
Summary
A new dual signal amplification strategy enables highly sensitive and specific detection of exosomal microRNAs (miRNAs) for early cancer diagnosis. This method uses enzyme-assisted DNA walkers and rolling circle amplification for accurate breast cancer detection in patients.
Area of Science:
- Biochemistry
- Molecular Biology
- Nanotechnology
Background:
- Exosomal microRNAs (miRNAs) are crucial biomarkers for cancer diagnosis, but their low abundance and short sequences challenge sensitive detection.
- Existing methods struggle with sensitivity and specificity, hindering early cancer detection and prognosis.
Purpose of the Study:
- To develop a novel dual signal amplification (DSA) strategy for highly sensitive, specific, and label-free detection of exosomal miRNAs.
- To establish a robust platform for early cancer diagnosis using exosomal miRNA analysis.
Main Methods:
- Integrated an APE1 enzyme-assisted DNA walker with rolling circle amplification (RCA) for signal amplification.
- Utilized magnetic beads for separation and Thioflavin T (ThT) for label-free fluorescence readout of G-quadruplex structures.
- Validated the DSA strategy using miR-10b as a breast cancer biomarker in cell-derived exosomes and human serum samples.
Main Results:
- Achieved a femtomolar detection limit (0.27 fM) with excellent specificity, discriminating single-nucleotide variants.
- Successfully differentiated miR-10b levels in exosomes from breast cancer cells versus normal cells.
- Demonstrated high diagnostic accuracy (AUC = 0.990) in distinguishing breast cancer patients from healthy controls based on serum exosomal miR-10b levels.
Conclusions:
- The DSA strategy offers a powerful tool for ultrasensitive and specific exosomal miRNA detection.
- This approach shows significant promise for advancing early cancer diagnosis and therapeutic development.
