Ultrasensitive Electrochemical DNA Biosensor Based on a Label-Free Assembling Strategy Using a Triblock polyA DNA Probe.,Analytical Chemistry

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Ultrasensitive Electrochemical DNA Biosensor Based on a Label-Free Assembling Strategy Using a Triblock polyA DNA Probe.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2019-12-05 , DOI: 10.1021/acs.analchem.9b04757
Lele Wang ₁ , Yanli Wen ₁ , Xue Yang ₁ , Li Xu ₁ , Wen Liang ₁ , Ying Zhu _{2,

3} , Lihua Wang _{2,

3} , Yan Li ₄ , Yuan Li ₁ , Min Ding ₁ , Shuzhen Ren ₁ , Zhenzhou Yang ₁ , Min Lv _{2,

3} , Jichao Zhang _{2,

3} , Kang Ma ₄ , Gang Liu ₁

Affiliation

Multiblock DNA probe attracted a large amount of scientific attention, for the development of multitarget biosensor and improved specificity/sensitivity. However, the development of multiblock DNA probes highly relied on the chemical synthesis of organic linkers or nanomaterials, which limited their practicability and biological compatibility. In this work, we developed a label-free assembling strategy using a triblock DNA capture probe, which connects two DNA probes with its intrinsic polyA fragment (probe-PolyA-probe, PAP). The middle polyA segment has a high affinity to the gold electrode surface, leading to excellent reproducibility, stability, and regeneration of our biosensor. Two flanking capture probes were tandemly co-assembled on the electrode surface with consistent spatial relationship and exactly the same amount. When combined with the target DNA, the hybridization stability was improved, because of the strong base stacking effect of two capture probes. The sensitivity of our biosensor was proved to be 10 fM, with a wide analysis range between 10 fM to 1 nM. Our PAP-based biosensor showed excellent specificity when facing mismatched DNA sequences. Even single nucleotide polymorphisms can be distinguished by each probe. The excellent practicability of our biosensor was demonstrated by analyzing genomic DNA both with and without PCR amplification.

中文翻译：

基于无标记组装策略的Triblock polyA DNA探针超灵敏电化学DNA生物传感器。

随着多靶点生物传感器的发展和特异性/敏感性的提高，多嵌段DNA探针引起了科学界的广泛关注。然而，多嵌段DNA探针的开发高度依赖于有机接头或纳米材料的化学合成，这限制了它们的实用性和生物相容性。在这项工作中，我们使用三嵌段DNA捕获探针开发了一种无标记的组装策略，该探针将两个DNA探针与其固有的polyA片段（探针-PolyA-探针，PAP）相连。中间的polyA段与金电极表面具有高亲和力，从而导致我们的生物传感器具有出色的重现性，稳定性和可再生性。两个侧翼捕获探针以一致的空间关系和完全相同的数量串联组装在电极表面上。当与靶DNA结合时，由于两个捕获探针的强碱基堆积效应，杂交稳定性得以提高。我们的生物传感器的灵敏度被证明为10 fM，分析范围在10 fM至1 nM之间。当面对错配的DNA序列时，我们基于PAP的生物传感器表现出出色的特异性。每个探针甚至可以区分单核苷酸多态性。我们的生物传感器具有出色的实用性，可通过分析有无PCR扩增的基因组DNA来证明。当面对错配的DNA序列时，我们基于PAP的生物传感器表现出出色的特异性。每个探针甚至可以区分单核苷酸多态性。我们的生物传感器具有出色的实用性，可通过分析有无PCR扩增的基因组DNA来证明。当面对错配的DNA序列时，我们基于PAP的生物传感器表现出出色的特异性。每个探针甚至可以区分单核苷酸多态性。我们的生物传感器具有出色的实用性，可通过分析有无PCR扩增的基因组DNA来证明。

更新日期：2019-12-06

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