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杂交诱导荧光增强的 DNA-Ag 纳米团簇/适体探针用于检测前列腺特异性抗原。

Hybridization induced fluorescence enhanced DNA-Ag nanocluster/aptamer probe for detection of prostate-specific antigen.

机构信息

Britton 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.

Britton 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.

出版信息

Colloids Surf B Biointerfaces. 2019 Mar 1;175:358-364. doi: 10.1016/j.colsurfb.2018.12.013. Epub 2018 Dec 7.

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 with the detection limit of 1.14 ng mL 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.

摘要

在这项工作中,提出了一种基于无标记银纳米簇(AgNC)的荧光探针来检测肿瘤标志物前列腺特异性抗原(PSA)。在实验中,使用包含与 PSA 适体不同部分互补片段的 DNA 序列来合成 DNA-Ag 纳米簇(DNA-AgNC)。获得的一些特定的 DNA-AgNC 在与 PSA 适体杂交后表现出显著的荧光增强。基于此,使用荧光猝灭法制备了一种简单的 DNA-AgNC/适体杂交探针用于 PSA 检测,因为 PSA 与其适体之间的特异性结合竞争抑制了 PSA 适体对 DNA-AgNC 的荧光增强效应。优化了模板 DNA 的序列、pH 值和结合缓冲液的盐浓度以及适体的浓度。在最佳条件下,可在 2-150ng/mL 范围内检测到 PSA,检测限为 1.14ng/mL(3σ;n=7)。该方法还成功应用于测定加标血清样品中的 PSA。众所周知,这是首次使用基于荧光 AgNC 的探针实现 PSA 检测的报道。这项工作将为构建基于 AgNC 的探针用于检测其他蛋白质提供参考。

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