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首页> 外文期刊>RSC Advances >Label-free electrochemical detection of DNA methyltransferase activity via a DNA tetrahedron-structured probe
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Label-free electrochemical detection of DNA methyltransferase activity via a DNA tetrahedron-structured probe

机译：通过DNA四面体结构探头通过DNA甲基转移酶活性的无标记电化学检测

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摘要

Recently, 3-dimensional DNA nanostructures have gained significant attention due to their programmable folding nature. Herein, a DNA tetrahedron-structured probe (DTP) is designed for the detection of methyltransferase (MTase) activity based on HRP-mimicking DNAzyme triggered deposition of polyaniline (PANI). In this strategy, MTase activity is measured with differential plus voltammetry (DPV) using PANI as the redox label. Furthermore, the DNA tetrahedron controlled the nanospacing between capture DNA, which greatly improves hybridization efficiency, MTase combination capability, and detection sensitivity. Using this method, more than 5 times higher signal gain is achieved compared with a single-strand capture DNA probe (ssP) without the tetrahedron structure. Under optimized experimental conditions, the DPV intensity is in linear correlation with M.SssI concentration ranging from 0.05 to 10 U mL ~(?1) , with a correlation coefficient of 0.9936 and a detection limit of 0.03 U mL ~(?1) (S/N = 3). The inhibition of 5-Aza- z ′-deoxycytidine or procaine on M.SssI activity is also evaluated. Good accuracy, precision and sensitivity are obtained when the method is applied in complex matrixes such as human serum samples, which is significant for clinical diagnosis and drug development.
机译：最近，由于其可编程的折叠性质，3维DNA纳米结构在显着的关注。这里，设计DNA四面体结构探针（DTP），用于检测基于HRP模拟的DNazyme触发聚苯胺（PANI）的沉积的甲基转移酶（MTase）活性。在该策略中，使用PANI作为氧化还原标记用差分加压伏安法（DPV）测量MTase活性。此外，DNA Tetrahedron控制捕获DNA之间的纳米间植物，这极大地提高了杂交效率，MTase组合能力和检测灵敏度。使用该方法，与没有四面体结构的单链捕获DNA探针（SSP）相比，实现了超过5倍的信号增益。在优化的实验条件下，DPV强度与0.05至10uml〜（α1）的M.SSI浓度为线性相关性，其相关系数为0.9936，检测限为0.03u ml〜（α1）（ s / n = 3）。还评估了抑制5-α-Z'-羟基胞苷或丙谷酮对M.SSSI活性的抑制作用。当该方法应用于诸如人血清样本的复杂基质中，获得良好的精度，精确和灵敏度，这对于临床诊断和药物发育是显着的。

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