DNA Nanosieve-Based Regenerative Electrochemical Biosensor Utilizing Nucleic Acid Flexibility for Accurate Allele Typing in Clinical Samples
DNA Nanosieve-Based Regenerative Electrochemical Biosensor Utilizing Nucleic Acid Flexibility for Accurate Allele Typing in Clinical Samples
复制标题
基于 DNA 纳米筛的再生电化学生物传感器利用核酸灵活性在临床样本中进行准确的等位基因分型
DOI：
10.1021/acssensors.0c02720
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发表时间：
2021-03-04
期刊：
影响因子：
8.9
通讯作者：
Xu, Xiong-Wei
中科院分区：
文献类型：
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作者：
Chen, Jin-Yuan;Yang, Liang-Yong;Xu, Xiong-Wei
在此，首次证明了具有区分ssDNA与dsDNA的能力的基于界面的DNA纳米筛。通过巯基DNA在金电极表面的自组装，可以很容易地构建DNA纳米筛，并且通过改变巯基DNA的浓度可以调节其空腔大小。使用[Ru（NH3）（6）]（3+）作为氧化还原剂的电化学计时库仑法显示，在1 μ M巯基-DNA-修饰的金电极中，探针与探针的平均分离为10.6 +/-0.3nm，使得具有类似于17 nm长度的刚性dsDNA不能渗透纳米筛，而随机卷曲的ssDNA由于其高柔性而可以进入纳米筛，这已经通过方波伏安法和亚甲蓝标记通过倒置杂交形式证明。在结合短DNA双链体的瞬时结合特性并引入再生探针（ssDNA的对应物）后，获得了高度可重复的基于纳米筛的E-DNA模型，在7个循环中的相对标准偏差（RSD）低至2.7%。最后，我们利用连接循环反应作为ssDNA扩增策略，构建了基于再生纳米筛的E-DNA传感器，实现了基于一个传感器的多个临床样本的连续测量，具有优异的等位基因分型性能。这项工作在生物传感器和生物芯片之间的低成本和高通量分析方面具有巨大的潜力，也为基于核酸柔性的DNA材料在DNA折纸和分子逻辑门中的未来应用开辟了新的途径。
Herein, an interface-based DNA nanosieve that has the ability to differentiate ssDNA from dsDNA has been demonstrated for the first time. The DNA nanosieve could be readily built through thiol-DNA's self-assembly on the gold electrode surface, and its cavity size was tunable by varying the concentration of thiol-DNAs. Electrochemical chronocoulometry using [Ru(NH3)(6)](3+) as redox revealed that the average probe-to-probe separation in the 1 mu M thiol-DNA-modified gold electrode was 10.6 +/- 0.3 nm so that the rigid dsDNA with a length of similar to 17 nm could not permeate the nanosieve, whereas the randomly coiled ssDNA could enter it due to its high flexibility, which has been demonstrated by square wave voltammetry and methylene blue labels through an upside-down hybridization format. After combining the transiently binding characteristic of a short DNA duplex and introducing a regenerative probe (the counterpart of ssDNA), a highly reproducible nanosieve-based E-DNA model was obtained with a relative standard deviation (RSD) as low as 2.7% over seven cycles. Finally, we built a regenerative nanosieve-based E-DNA sensor using a ligation cycle reaction as an ssDNA amplification strategy and realized one-sensor-based continuous measurement to multiple clinical samples with excellent allele-typing performance. This work holds great potential in low-cost and high-throughput analysis between biosensors and biochips and also opens up a new avenue in nucleic acid flexibility-based DNA materials for future applications in DNA origami and molecular logic gates.