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Label-free electrochemiluminescent detection of transcription factors with hybridization chain reaction amplification

机译：杂交链反应扩增的无标记电化学检测转录因子

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Because of the intrinsic importance of transcription factors (TFs) as targets in clinical diagnosis and drug development, the simple and sensitive detection of TFs is very essential for biological studies and medical diagnostics. However, most of the reported methods involve complicated operations or labelling processes and in addition, an electrochemiluminescence (ECL) method with various distinct advantages hasn't been developed for TF detection until now. In this work, we describe a novel simple and efficient strategy for label-free ECL detection of transcription factors with hybridization chain reaction (HCR) signal amplification. Based on the Ag ~(+) -stabilized self-assembly triplex DNA, in the presence of TFs, TFs specifically bond to the duplex DNA (dsDNA) recognition probes, resulting in the separation of target DNA from the triplex structure. With the SH-capture probe DNA assembled gold electrode, the presence of the target DNA and helper DNA-1 and helper DNA-2 leads to the formation of long chain dsDNA polymers on the gold electrode surface through hybridization chain reaction, which allows the intercalation of numerous ECL indicators Ru(phen) _(3) ~(2+) (phen = 1,10-phenanthroline) into the dsDNA grooves, resulting in significantly amplified ECL signal output. The proposed strategy combines the amplification power of the HCR and the inherent high sensitivity of the ECL technique, resulting in the sensitive detection of transcription factors with a detection limit of as low as 0.017 nM and a broad dynamic range from 0.05 to 2 nM. The distinct advantage of the method is that it is label-free, has high sensitivity and requires no separation of the signal generation strand, which boosts the potential application for real sample detection.
机译：由于转录因子（TFS）作为临床诊断和药物发育的目标的内在重要性，TFS的简单和敏感性检测对于生物学研究和医学诊断至关重要。然而，大多数报道的方法涉及复杂的操作或标记过程，另外，直到现在，尚未开发具有各种不同优点的电化学发光（ECL）方法。在这项工作中，我们描述了一种新的简单有效的无标记ECL检测与杂交链反应（HCR）信号放大的转录因子的简单有效策略。基于Ag〜（+）-Stabilized自组装三重双链DNA，在TFS存在下，TFS特别键入双相DNA（DSDNA）识别探针，导致靶DNA与三重结构的分离。通过SH捕获探针DNA组装金电极，靶DNA和辅助DNA-1和辅助DNA-2的存在导致通过杂交链反应形成在金电极表面上的长链DSDNA聚合物，这允许插入许多ECL指标ru（phen）_（3）〜（2+）（phen = 1,10-菲啉）进入DSDNA槽，导致显着放大的ECL信号输出。所提出的策略结合了HCR的扩增功率和ECL技术的固有的高灵敏度，导致检测因子的敏感因子的检测限度为0.017nm，宽动态范围为0.05至2nm。该方法的明显优点是它无标记，具有高灵敏度，并且不需要分离信号产生股线，这促进了真实样本检测的潜在应用。

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