Ultrasensitive Optical DNA Biosensor Based on Surface Immobilization of Molecular Beacon by a Bridge Structure.

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  • LI Jun
    Institute of Biotechnology and Biomedical Engineering, College of Chemistry & Chemical Engineering, Hunan University
  • TAN Weihong
    Institute of Biotechnology and Biomedical Engineering, College of Chemistry & Chemical Engineering, Hunan University
  • WANG Kemin
    Institute of Biotechnology and Biomedical Engineering, College of Chemistry & Chemical Engineering, Hunan University
  • XIAO Dan
    Institute of Biotechnology and Biomedical Engineering, College of Chemistry & Chemical Engineering, Hunan University
  • YANG Xiaohai
    Institute of Biotechnology and Biomedical Engineering, College of Chemistry & Chemical Engineering, Hunan University
  • HE Xiaoxiao
    Institute of Biotechnology and Biomedical Engineering, College of Chemistry & Chemical Engineering, Hunan University
  • TANG Zhiwen
    Institute of Biotechnology and Biomedical Engineering, College of Chemistry & Chemical Engineering, Hunan University

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Abstract

A novel biotinylated molecular beacon (MB) probe was developed to prepare a DNA biosensor using a bridge structure. MB was biotinylated at the quencher side of the stem and linked on a biotinylated glass cover slip through streptavidin, which acted as a bridge between MB and glass matrix. An efficient fluorescence microscope system was constructed to detect the fluorescence change caused by the conformation change of MB in the presence of complementary DNA target. The proposed biosensor was used to directly detect, in real-time, the target DNA molecules. The bridge immobilization method caused the proposed DNA biosensor to have a faster and more stable response. Under the optimal conditions, the newly developed DNA biosensor showed a linear response toward ssDNA in the range of 5 - 100 nM with a detection limit of 2 nM. It was interesting to note that the described biosensor was reproducible after being regenerated by urea.

Journal

  • Analytical Sciences

    Analytical Sciences 17 (10), 1149-1153, 2001

    The Japan Society for Analytical Chemistry

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