Hubei Collaborative Innovation Center for Advanced Organic Chemical Materials, Ministry of Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules & College of Chemistry and Chemical Engineering, Hubei University, Wuhan 430062, PR China.
Hubei Collaborative Innovation Center for Advanced Organic Chemical Materials, Ministry of Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules & College of Chemistry and Chemical Engineering, Hubei University, Wuhan 430062, PR China.
Biosens Bioelectron. 2016 Mar 15;77:451-6. doi: 10.1016/j.bios.2015.09.068. Epub 2015 Nov 14.
A specific and sensitive method was developed for quantitative detection of miRNA by integrating horseradish peroxidase (HRP)-assisted catalytic reaction with a simple electrochemical RNA biosensor. The electrochemical biosensor was constructed by a double-stranded DNA structure. The structure was formed by the hybridization of thiol-tethered oligodeoxynucleotide probes (capture DNA), assembled on the gold electrode surface, with target DNA and aminated indicator probe (NH2-DNA). After the construction of the double-stranded DNA structure, the activated carboxyl groups of graphene quantum dots (GQDs) assembled on NH2-DNA. GQDs were used as a new platform for HRP immobilization through noncovalent assembly. HRP modified biosensor can effectively catalyze the hydrogen peroxide (H2O2)-mediated oxidation of 3,3',5,5'-tetramethylbenzidine (TMB), accompanied by a change from colorless to blue in solution color and an increased electrochemical current signal. Due to GQDs and enzyme catalysis, the proposed biosensor could sensitively detect miRNA-155 from 1 fM to 100 pM with a detection limit of 0.14 fM. High performance of the biosensor is attributed to the large surface-to-volume ratio, excellent compatibility of GQDs. For these advantages, the proposed method holds great potential for analysis of other interesting tumor makers.
本研究构建了一种基于双链 DNA 结构的电化学 RNA 生物传感器,通过辣根过氧化物酶(HRP)辅助催化反应与简单的电化学 RNA 生物传感器相结合,实现了 miRNA 的定量检测。该生物传感器由巯基修饰的寡脱氧核苷酸探针(捕获 DNA)组装在金电极表面形成,探针与目标 DNA 和氨基化指示剂探针(NH2-DNA)杂交,形成双链 DNA 结构。在构建双链 DNA 结构后,将石墨烯量子点(GQDs)的活化羧基基团组装在 NH2-DNA 上。GQDs 通过非共价组装被用作 HRP 固定化的新平台。HRP 修饰的生物传感器可以有效地催化过氧化氢(H2O2)介导的 3,3',5,5'-四甲基联苯胺(TMB)氧化,溶液颜色由无色变为蓝色,电化学电流信号增强。由于 GQDs 和酶的催化作用,该生物传感器可以从 1 fM 到 100 pM 范围内灵敏地检测 miRNA-155,检测限为 0.14 fM。该生物传感器具有优异的性能,这归因于 GQDs 的大表面积与体积比和良好的兼容性。鉴于这些优点,该方法有望用于分析其他感兴趣的肿瘤标志物。