Bingyuan Fan, Qian Wang, Shan Wang, Yahui Gao, Yan Liang, Jinru Pan, Xinrui Fu, Li Li, Wei Meng
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引用次数: 0
摘要
MiR-378 在肝细胞癌、肾细胞癌和非小细胞肺癌等多种癌症中异常表达。在此,我们开发了一种基于外切酶 III(Exo III)的无标记、无固定化的比率计量均相电化学策略,用于简便快速地测定 miR-378。该方法设计了两个 3′突起的发夹 DNA 探针(HPs)。多柔比星(DOX)和亚铁氰化钾(Fe2+)被用作无标记探针,在溶液阶段产生响应信号(IDOX)和参考信号(IFe2+)。当溶液中没有靶标存在时,HP 稳定,大部分 DOX 被夹杂在 HP 的茎中,DOX 的扩散速率显著降低,导致电化学信号响应降低。当存在 miR-378 时,由 Exo III 裂解引发的双循环信号放大开始启动,最终破坏了 HP1 和 HP2 的发夹结构,向溶液中释放出大量 DOX,产生更强的电化学信号,低至 50 pM。这种检测方法具有极佳的选择性,在生物系统中具有很高的应用潜力,并为 miR-378 的电化学检测提供了简单、低成本的方法。
Label-Free Ratiometric Homogeneous Electrochemical Strategy Based on Exonuclease III-Aided Signal Amplification for Facile and Rapid Detection of miR-378
MiR-378 is abnormally expressed in various cancers, such as hepatocellular carcinoma, renal cell carcinoma, and nonsmall cell lung cancer. Here, we developed a label- and immobilization-free ratiometric homogeneous electrochemical strategy based on exonuclease III (Exo III) for the facile and rapid determination of miR-378. Two 3′-protruding hairpin DNA probes (HPs) are designed in this strategy. Doxorubicin (DOX) and potassium ferrocyanide (Fe2+) were used as label-free probes to produce a response signal (IDOX) and a reference signal (IFe2+) in the solution phase. When no target was present in the solution, the HP was stable, most of the DOX was intercalated in the stem of the HP, and the diffusion rate of DOX was significantly reduced, resulting in reduced electrochemical signal response. When miR-378 was present, double-cycle signal amplification triggered by Exo III cleavage was initiated, ultimately disrupting the hairpin structures of HP1 and HP2 and releasing a large amount of DOX into the solution, yielding a stronger electrochemical signal, which was low to 50 pM. This detection possesses excellent selectivity, demonstrating high application potential in biological systems, and offers simple and low-cost electrochemical detection for miR-378.
期刊介绍:
International Journal of Analytical Chemistry publishes original research articles that report new experimental results and methods, especially in relation to important analytes, difficult matrices, and topical samples. Investigations may be fundamental, or else related to specific applications; examples being biological, environmental and food testing, and analysis in chemical synthesis and materials processing.
As well as original research, the International Journal of Analytical Chemistry also publishes focused review articles that examine the state of the art, identify emerging trends, and suggest future directions for developing fields.