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通过将导电流体磁珠与二茂铁修饰的金纳米粒子/链霉亲和素偶联物相结合,对脑胶质瘤患者血清样本中的 miRNA 进行放大伏安检测。

Amplified voltammetric detection of miRNA from serum samples of glioma patients via combination of conducting magnetic microbeads and ferrocene-capped gold nanoparticle/streptavidin conjugates.

机构信息

College of Chemistry and Chemical Engineering, Central South University, Changsha, Hunan 410083, PR China.

SunYat-Sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, Guangdong 510060, PR China.

出版信息

Biosens Bioelectron. 2016 Dec 15;86:502-507. doi: 10.1016/j.bios.2016.07.010. Epub 2016 Jul 5.

Abstract

MicroRNA (miRNA) plays a key regulatory role in many biological processes, emerging as an important biomarker for a large variety of cancer diseases. Employing gold nanoparticle (AuNP)-coated magnetic microbeads (AuNP-MMBs) as an immobilization matrix for higher loading density of hairpin-structured DNA probes and then ferrocene (Fc)-capped gold nanoparticle/streptavidin conjugates, amplified electrochemical assay of miRNA has been performed. In the presence of target miRNA, a novel assembly was formed via linking biotinylated hairpin DNA probe-covered AuNP-MMBs with Fc-capped gold nanoparticle/streptavidin conjugates and then collected by magnetic electrodes for voltammetric detection. The enlarged surface area, good conductivity of AuNP-MMBs and the multiple Fc tags on the electrode surface ensure high sensitivity of the method. The oxidation peak current of Fc tags is proportional to the concentrations of miRNA ranging from 5 fM to 100 fM, and a detection limit of 0.14 fM was achieved. The proposed assay is highly selective and reproducible, serving as a viable alternative for the detection of miRNA-182 from serum samples of glioma patients.

摘要

微小 RNA(miRNA)在许多生物过程中发挥着关键的调节作用,是多种癌症疾病的重要生物标志物。本研究采用金纳米颗粒(AuNP)包覆的磁性微球(AuNP-MMBs)作为发夹结构 DNA 探针的固定基质,实现了 miRNA 的高载量电化学放大检测。在靶 miRNA 的存在下,通过连接生物素化发夹 DNA 探针覆盖的 AuNP-MMBs 和 Fc 封端的金纳米颗粒/链霉亲和素缀合物,形成了一种新的组装体,并通过磁电极收集进行伏安检测。AuNP-MMBs 的大表面积、良好的导电性和电极表面上的多个 Fc 标记确保了该方法的高灵敏度。Fc 标记的氧化峰电流与 miRNA 的浓度呈正比,范围从 5 fM 到 100 fM,检测限达到 0.14 fM。该方法具有高度的选择性和重现性,可作为检测神经胶质瘤患者血清样本中 miRNA-182 的一种可行方法。

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