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化学学报
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化学学报 ›› 2011, Vol. 69 ›› Issue (02): 183-189. 上一篇    下一篇

研究论文

高灵敏选择性检测凝血酶的适体纳米银共振散射光谱探针

黄世稳1,2，李纪顺1，梁爱惠1，蒋治良*,1   

  1. (1广西师范大学环境与资源学院 珍稀濒危动植物生态与环境保护省部共建教育部重点实验室 桂林 541004)
    (2右江民族医学院 百色 533000)
  • 投稿日期:2010-08-05 修回日期:2010-09-24 发布日期:2010-09-27
  • 通讯作者: 蒋治良 E-mail:zljiang@mailbox.gxnu.edu.cn
  • 基金资助:

    国家自然科学基金项目

Highly Sensitive and Selective Detection of Thrombin by Aptamer- modified Nanosilver Resonance Scattering Spectral Probe

Huang Shiweng1,2 Li Jishun1 Liang Aihui1 Jiang Zhiliang*,1   

  1. (1 Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection, Ministry of Education, School of Environment and Resource Sciences, Guangxi Normal University, Guilin 541004)
    (2 Youjiang Medical University for Nationalities, Baise 533000)
  • Received:2010-08-05 Revised:2010-09-24 Published:2010-09-27

PDF

1421

被引次数

36

采用四氢硼钠制备了较稳定的纳米银, 并用凝血酶(TB)适体修饰纳米银制备了识别凝血酶的适体纳米银探针. 在pH 7.0的Tris-HCl缓冲溶液及KCl存在下, 适体纳米银探针与凝血酶特异结合生成G-四分体和纳米银聚集体, 导致体系在480 nm处的共振散射峰增强. 随着凝血酶浓度的增大, 生成的纳米银聚集体越多, 共振散射强度线性增强. 采用不同银金物质的量之比的银金纳米微粒制备其探针并用于凝血酶的共振散射光谱分析. 结果表明, 适体纳米银探针体系的灵敏度较高、线性范围较宽, 且纳米银制备较简便, 成本低廉, 其共振散射增强值ΔI480 nm与凝血酶浓度在0.14～11.5 nmol/L范围内存在良好的线性关系, 检出限为17 pmol/L凝血酶.

关键词: 纳米银, 适体, 凝血酶, 共振散射光谱法

Silver nanoparticles were prepared by NaBH4 reduction procedure. An aptamer-modified Ag nanoprobe (AgSH-ssDNA) for thrombin was prepared. In the media of pH 7.0 Tris-HCl and KCl, the AgSH-ssDNA specifically combined with thrombin to form G-quadruplex and Ag cluster, which resulted in the resonance scattering intensity at 480 nm (I480 nm) increasing. As the amount of thrombin increased, the amount of Ag clusters increased, and the I480 nm value increased. At the same time, Ag/Au nanoparticles of different Ag/Au molar ratio were also prepared for the thrombin RS analysis. The results showed the Ag system had some advantages such as high sensitivity, wide linear range, low-cost, and the simplicity of nanoparticles preparation. Its increased intensity ΔI480 nm was linear to the thrombin concentration in the range of 0.14～11.5 nmol/L, with a regression equation of ΔI480 nm＝19.1C＋2.2, a correlation coefficient of 0.9833, and a detection limit of 17 pmol/L. This method was applied to detect thrombin in human plasma samples with satisfactory results.

Key words: nanosilver, aptamer, thrombin, resonance scattering spectral assay