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电化学(中英文) ›› 2019, Vol. 25 ›› Issue (2): 232-243.  doi: 10.13208/j.electrochem.181051

• 研究论文 • 上一篇    下一篇

基于催化发卡自组装和 Ru(NH3)63+ 的核酸光电化学灵敏分析

付亚敏,闫小霞,张小华*,陈金华*   

  1. 湖南大学化学化工学院,化学生物传感与计量学国家重点实验室,湖南 长沙410082
  • 收稿日期:2018-12-18 修回日期:2019-01-09 出版日期:2019-04-28 发布日期:2019-02-01
  • 通讯作者: 张小华,陈金华 E-mail: mickyxie@hnu.edu.cn; chenjinhua@hnu.edu.cn

Sensitive Photoelectrochemical Assay of Nucleic Acids Based on Catalytic Hairpin Assembly and Ru(NH3)63+

FU Ya-min, YAN Xiao-xia, ZHANG Xiao-hua*, CHEN Jin-hua*   

  1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, Hunan, China
  • Received:2018-12-18 Revised:2019-01-09 Published:2019-04-28 Online:2019-02-01
  • Contact: ZHANG Xiao-hua, CHEN Jin-hua E-mail: mickyxie@hnu.edu.cn; chenjinhua@hnu.edu.cn
  • Supported by:
    This work was financially supported by the National Science Foundation of China (Grants Nos.21727810 and 21475035) and the Foundation for Innovative Research Groups of NSFC (21521063)

摘要: 基于催化发卡自组装反应(CHA)和电活性材料[Ru(NH3)6]Cl3,发展了一种“信号增强”型光电化学生物传感器,实现了核酸的灵敏检测. 首先,采用逐层离子吸附法(SILAR)将CdS 固定于TiO2/ITO 电极表面. 光电材料CdS 不仅能够将TiO2 的吸收范围从紫外光区拓展到可见光区,而且还能提高光电转换效率. 之后,通过Cd-S 键将捕获DNA(C-DNA)固定于CdS/ TiO2/ITO 电极表面. 与此同时,将Au 结合的发卡DNA 探针1(Au-HP1),发卡DNA 探针2(HP2)和目标DNA(T-DNA)混合物于溶液中进行CHA 反应,得到大量的Au-HP1:HP2 复合物. 再通过Au-HP1:HP2 复合物与C-DNA 的杂交反应将大量的双链DNA 引入到电极表面. 最后,将电活性物质Ru(NH3)63+嵌入DNA 的磷酸骨架中,从而使得光电流大幅度的增强. 该光电生物传感器检测核酸的线性范围为10 fmol·L-1 到 1500 fmol·L-1,检测线为6.19 fmol·L-1,在生物分析、新药筛选以及疾病的早期诊断等方面具有潜在的应用前景.

关键词: 光电化学生物传感器, 核酸, 催化发卡自组装, [Ru(NH3)6]Cl3, CdS, TiO2

Abstract: A simple “signal-on” photoelectrochemical (PEC) sensing platform for sensitive assay of nucleic acids was developed by coupling catalytic hairpin assembly (CHA) signal amplification strategy with Ru(NH3)63+. Herein, cadmium sulfide (CdS) was deposited on the TiO2/indium tin oxide (ITO) electrode by a method of successive ionic layer adsorption and reaction (SILAR), serving as one kind of photoelectric material to broaden absorption range of TiO2 and to improve the photoelectric conversion efficiency. Thereafter, the capture DNA (C-DNA) was immobilized on the CdS/TiO2/ITO electrode. Simultaneously, Au-hairpin DNA probe 1 (Au-HP1) and hairpin DNA probe 2 (HP2) were able to hybridize to produce many Au-HP1:HP2 complexes under the existence of target DNA (T-DNA) based on CHA process. After that, C-DNA could capture Au-HP1:HP2 complex, leading to lots of double-stranded DNAs on the electrode to load numerous Ru(NH3)63+, which resulted in a remarkable increase of photocurrent. As a result, a wide linear range (10 fmol·L-1 to 1500 fmol·L-1) and a low detection limit (6.19 fmol·L-1) toward T-DNA were achieved. The developed method would have great potential applications in bioanalysis, screening of new drugs and early disease diagnosis.

Key words: photoelectrochemical biosensor, nucleic acid, catalytic hairpin assembly, [Ru(NH3)6]Cl3, CdS, TiO2

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