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Updated: Jan 9, 2026

Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis
Published on: July 8, 2025
Hybridization induced fluorescence enhanced DNA-Ag nanocluster/aptamer probe for detection of prostate-specific
Bi-Yun Fang1, Jie An2, Bo Liu3
1Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics - Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, Hubei, PR China; Key Laboratory of Biomedical Photonics (HUST), Ministry of Education, Huazhong University of Science and Technology, Wuhan, 430074, Hubei, PR China; Institute of Medical Engineering, Department of Biophysics, School of Basic Medical Sciences, Health Science Center, Xi'an Jiaotong University, Xi'an, Shanxi, 710061, PR China.
Abstract:
In this work, a label-free Ag nanocluster (AgNC)-based fluorescent probe is proposed to detect tumor marker, prostate-specific antigen (PSA). In the experiments, DNA sequences containing segments complemented to different parts of PSA aptamer were used to synthesize DNA-Ag nanoclusters (DNA-AgNC). Some of the obtained specific DNA-AgNC exhibited significant fluorescence increase after hybridization with PSA aptamer. Based on this, a simple DNA-AgNC/aptamer hybridization probe was fabricated for PSA detection using fluorescence quenching, because competitively specific binding between PSA and its aptamer inhibited the fluorescence enhancement effect of PSA aptamer on DNA-AgNC. The sequence of template DNA, pH and salt concentration of binding buffer, and the concentration of aptamer were optimized. Under optimum conditions, the concentration of PSA within the range of 2-150 ng mL-1 with the detection limit of 1.14 ng mL-1 was detected (3σ; n = 7). This approach was also successfully applied to determine PSA in spiked serum samples. As is well known, this was the first report to realize PSA detection using fluorescent AgNC-based probe. This work would provide reference for construction of AgNC-based probes for detecting other proteins.
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