Liu Xiujuan, Zhang Meixiang, Chen Ze, Cui Jiuqing, Yang Long, Lu Zihe, Qi Fang, Wang Haixia
Department of Intensive Care Unit, The First Hospital of Qinhuangdao, Qinhuangdao, China.
Department of Intensive Care Unit, Hebei Petrochina Central Hospital, Langfang, China.
Front Bioeng Biotechnol. 2021 Dec 13;9:799370. doi: 10.3389/fbioe.2021.799370. eCollection 2021.
MicroRNA (miRNA) detection has attracted widespread interest as a tumor detection marker. In this work, a miRNA-responsive visual and temperature sensitive probe composed of a horseradish peroxidase (HRP)-encapsulated DNA hydrogel was designed and synthesized. The biosensor converted the miRNA hybridization signal to a photothermal effect which was measured using a digital thermometer. The substrate DNA linker strand of the hydrogel hybridizes with different sequences of miRNA resulting in the collapse of the hydrogel and the release of HRP. HRP oxidizes 3,3',5,5'-tetramethylbenzidine (TMB) resulting in a color change and a strong photothermal effect was observed after shining near-infrared light on the oxidized product. The thermometer-based readout method has a wide linear range (0.5-4.0 µM) and a limit of detection limit of 7.8 nM which is comparable with traditional UV-vis absorption spectrometry detection and quantitative real time polymerase chain reaction methods. The low cost, ease of operation, and high sensitivity shows that this biosensor has potential for point-of-care biomolecular detection and biomedical applications.
微小RNA(miRNA)检测作为一种肿瘤检测标志物已引起广泛关注。在这项工作中,设计并合成了一种由辣根过氧化物酶(HRP)封装的DNA水凝胶组成的miRNA响应型视觉和温度敏感探针。该生物传感器将miRNA杂交信号转化为光热效应,并用数字温度计进行测量。水凝胶的底物DNA连接链与不同序列的miRNA杂交,导致水凝胶塌陷并释放HRP。HRP氧化3,3',5,5'-四甲基联苯胺(TMB)导致颜色变化,并且在近红外光照射氧化产物后观察到强烈的光热效应。基于温度计的读出方法具有宽线性范围(0.5 - 4.0 μM)和7.8 nM的检测限,与传统紫外可见吸收光谱检测和定量实时聚合酶链反应方法相当。低成本、易于操作和高灵敏度表明该生物传感器在即时护理生物分子检测和生物医学应用方面具有潜力。