Label-free and sensitive strategy for microRNAs detection based on the formation of boronate ester bonds and the dual-amplification of gold nanoparticles.
Label-free and sensitive strategy for microRNAs detection based on the formation of boronate ester bonds and the dual-amplification of gold nanoparticles.
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DOI：
10.1016/j.bios.2013.03.074
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发表时间：
2013-09
影响因子：
12.6
通讯作者：
N. Xia;Liping Zhang;Guifang Wang;Qingqin Feng;Lin Liu
中科院分区：
文献类型：
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作者：
N. Xia;Liping Zhang;Guifang Wang;Qingqin Feng;Lin Liu
microRNAs（miRNAs）通过翻译抑制或降解信使RNA来调节基因表达，是肿瘤诊断和预后的重要分子生物标志物。因此，迫切需要简单，灵敏和具有成本效益的miRNAs检测方法。RNA与DNA结构的主要区别在于RNA中核糖的2 ′位存在羟基，这使得RNA分子在链的末端含有顺式二醇。众所周知，亲水性硼酸与顺式二醇形成共价键。在这项工作中，我们报告了一种无标记的和敏感的方法检测的miRNA的基础上形成的硼酸酯共价键和双扩增的金纳米粒子（AuNPs）。具体而言，miRNA被预先固定的DNA探针捕获在金电极上，并通过在MBA-AuNP的硼酸和miRNA的二醇之间形成紧密的共价键而用4-巯基苯硼酸（MBA）封端的AuNP（MBA-AuNP）衍生。电化学活性多巴胺（DA）封端的金纳米颗粒（DA-AuNPs）通过硼酸和DA标签的相互作用被锚定的MBA-AuNPs连接，这有助于miRNA的放大伏安检测。分析优点（例如，灵敏度、再现性、储存稳定性、动态范围和选择性）。我们相信，这一结果将是有价值的生物传感器的发展，用于检测生物基质中的miRNA。
MicroRNAs (miRNAs), regulating gene expression by translational repression or degradation of messenger RNAs, are believed to be important for cancer diagnosis and prognosis serving as reliable molecular biomarkers. Simple, sensitive, and cost-effective assays for miRNAs are therefore in urgent demand. The main difference in the structure of RNA versus DNA is the presence of a hydroxyl group at the 2′ position of the ribose sugar in RNA, which makes the RNA molecule contain cis-diol at the end of the chain. Hydrophilic boronic acids are well known to form covalent bonds with cis-diols. In this work, we reported a label-free and sensitive method for the detection of miRNAs based on the formation of boronate ester covalent bonds and the dual-amplification of gold nanoparticles (AuNPs). Specifically, miRNAs were captured by the pre-immobilized DNA probes at the gold electrode, and derivatized with 4-mercaptophenylboronic acid (MBA)-capped AuNPs (MBA-AuNPs) through the formation of tight covalent bonds between the boronic acids of MBA-AuNPs and diols of miRNAs. Electrochemically active dopamine (DA)-capped AuNPs (DA-AuNPs) were then attached by the anchored MBA-AuNPs via the interaction of boronic acids and DA tags, which facilities the amplified voltammetric detection of miRNAs. Analytical merits (e.g., sensitivity, reproducibility, storage stability, dynamic range, and selectivity) were addressed. We believe that the results will be valuable for the development of biosensor for the detection of miRNAs in a biological matrix.