Department of Chemistry, Liaocheng University, Liaocheng, Shandong 252000, China.
Department of Chemistry, Liaocheng University, Liaocheng, Shandong 252000, China.
Talanta. 2018 Mar 1;179:414-419. doi: 10.1016/j.talanta.2017.11.036. Epub 2017 Nov 20.
In the present work a highly sensitive and selective aptasensor was developed for the determination of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) based on the hybridization chain reaction (HCR) signal amplification. It was observed that the aptamer of 8-OH-dG could hybridize with the capture DNA immobilized on the gold electrode with a sticky tail left, which initiated the HCR and led to the formation of extended dsDNA structure on the electrode surface. Then the electroactive species ([Ru(NH)], RuHex) intercalated into the dsDNA grooves to generate the amplified signal. However, in the presence of 8-OH-dG, the aptamer containing G-rich nucleic acid sequences would be induced to form a G-quadruplex structure, which made it impossible to continue the HCR. So the detection signal will significantly decrease. Under the optimal conditions, the peak current of RuHex was linear with the logarithm of 8-OH-dG concentration in the range from 10pM to 100μM with the detection limit of 2.5pM. By integrating the merits of enzyme-free amplification power of the HCR and the inherent high sensitivity of the electrochemical technique, the prepared aptasensor not only showed high sensitivity for the detection of 8-OH-dG, but also exhibited good selectivity against to the uric acid, an important interferent in the urine sample. Particularly, the aptasensor was applied to detect 8-OH-dG in urine samples with satisfactory results.
在本工作中,基于杂交链式反应(HCR)信号放大,开发了一种用于测定 8-羟基-2'-脱氧鸟苷(8-OH-dG)的高灵敏度和选择性适体传感器。结果表明,8-OH-dG 的适体能与固定在金电极上的捕获 DNA 杂交,留下粘性尾巴,从而引发 HCR,并导致电极表面形成扩展的 dsDNA 结构。然后,将电化学活性物质([Ru(NH)] ,RuHex)插入 dsDNA 沟槽中以产生放大信号。然而,在存在 8-OH-dG 的情况下,含有富含 G 的核酸序列的适体会被诱导形成 G-四链体结构,从而无法继续进行 HCR。因此,检测信号会显著降低。在最佳条件下,RuHex 的峰电流与 8-OH-dG 浓度的对数在 10pM 至 100μM 的范围内呈线性关系,检测限为 2.5pM。通过整合 HCR 的无酶扩增功率和电化学技术固有的高灵敏度的优点,制备的适体传感器不仅对 8-OH-dG 的检测具有高灵敏度,而且对尿酸(尿液样品中的重要干扰物)具有良好的选择性。特别是,该适体传感器被应用于检测尿液样品中的 8-OH-dG,结果令人满意。