Dendritic nucleic acid probes for DNA biosensors
Dendritic nucleic acid probes for DNA biosensors
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DOI：
10.1021/ja980619p
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发表时间：
1998-08-19
影响因子：
15
通讯作者：
Getts, RC
中科院分区：
文献类型：
--
作者：
Wang, J;Jiang, M;Getts, RC
DNA杂交生物传感器在遗传和病原性疾病的快速诊断方面具有很大的应用前景。1-3这种装置通常依赖于单链寡核苷酸探针的固定，该探针通过杂交选择性地识别其互补的靶序列。到目前为止，大多数活动都集中在不同转导模式的开发上，而对DNA识别过程的关注很少。在本文中，我们报告了使用大分子核酸结构，如DNA树状大分子4-6或支链DNA7作为DNA生物传感器的识别元件所产生的优点。为此，我们使用了DNA树枝状大分子，这种高度分枝的结构具有多个单链臂，能够与互补的核酸序列杂交。5将这些超支化的球形结构固定在物理传感器上，极大地提高了表面的杂交能力，从而提高了DNA生物传感器的灵敏度和扩展的线性。DNA树状大分子通过设计的DNA组分的顺序杂交来组装，每一代都以受控的指数增长。5、8据我们所知，以前没有人尝试使用核酸树枝状大分子来创建DNA生物传感器。含有多个杂交位点的分枝(梳状)或树枝状DNA结构以前曾被用于提高溶液相核酸印迹分析的灵敏度。7，8与三维DNA树状大分子相比，前者受DNA合成的化学限制仅限于较少的杂交位点。在本工作中，我们使用质量敏感的压电换能器，9与表面受限的树枝状大分子，以证明提高的杂交能力和检测能力，并直接(没有指示剂)监测杂交动力学。为此，我们使用吸附和电聚合物(多酚)包埋的方法固定第四代(G4)。
DNA hybridization biosensors hold great promise for the rapid diagnosis of genetic and pathogenic diseases. 1-3 Such devices commonly rely on the immobilization of a single-stranded oligonucleotide probe that selectively recognizes its complementary target sequence through hybridization. To date, most activity has focused on the development of different transduction modes, with little attention given to the DNA recognition process. In this paper, we report on the advantages accrued from the use of macromolecular nucleic acid structures such as DNA dendrimers4-6 or branched DNA7 as the recognition elements in DNA biosensors. For this purpose we used DNA dendrimers which are highly branched structures possessing multiple single-stranded arms capable of hybridizing with a complementary nucleic acid sequence. 5 The immobilization of these hyperbranched spherical structures onto physical transducers greatly increases the hybridization capacity of the surface and, hence, leads to enhanced sensitivity and extended linearity of DNA biosensors. DNA dendrimers are assembled via sequential hybridization of designed DNA components, with a controlled exponential growth with each successive generation. 5, 8 To our knowledge, no previous attempts have been made to use nucleic acid dendrimers for the creation of DNA biosensors. Branched (comband fork-) or dendritic DNA structures, containing multiple hybridization sites, have been used previously for enhancing the sensitivity of solution-phase nucleic acid blot assays. 7, 8 The former are restricted by the chemistry of the DNA synthesis to fewer hybridization sites when compared to three-dimensional DNA dendrimers.In the present work, we employed mass-sensitive piezoelectric transducers, 9 with surface-confined dendrimers, to demonstrate the increased hybridization capacity and detection capability and to monitor directly (without an indicator) the kinetics of hybridization. For this purpose we used adsorption and electropolymeric (polyphenol) entrapment for immobilizing the 4-generation (G4)