Shandong Sino-Japanese Center for Collaborative Research of Carbon Nanomaterials, College of Chemistry and Chemical Engineering, Qingdao University, Qingdao 266071, China.
Shandong Provincial Key Laboratory of Detection Technology for Tumor Markers, College of Chemistry and Chemical Engineering, Linyi University, Linyi 276005, China.
Sensors (Basel). 2018 Dec 26;19(1):77. doi: 10.3390/s19010077.
Basing on the conformation change of aptamer caused by proteins, a simple and sensitive protein fluorescent assay strategy is proposed, which is assisted by the isothermal amplification reaction of polymerase and nicking endonuclease. In the presence of platelet-derived growth factor (PDGF-BB), the natural conformation of a DNA aptamer would change into a Y-shaped complex, which could hybridize with a molecular beacon (MB) and form a DNA duplex, leading to the open state of the MB and generating a fluorescence signal. Subsequently, with further assistance of isothermal recycling amplification strategies, the designed aptamer sensing platform showed an increment of fluorescence. As a benefit of this amplified strategy, the limit of detection (LOD) was lowered to 0.74 ng/mL, which is much lower than previous reports. This strategy not only offers a new simple, specific, and efficient platform to quantify the target protein in low concentrations, but also shows a powerful approach without multiple washing steps, as well as a precious implementation that has the potential to be integrated into portable, low-cost, and simplified devices for diagnostic applications.
基于适配体因蛋白质而发生的构象变化，提出了一种简单、灵敏的蛋白质荧光检测策略，该策略辅助了聚合酶和切口酶的等温扩增反应。在血小板衍生生长因子（PDGF-BB）存在的情况下，DNA 适配体的天然构象会变成 Y 形复合物，与分子信标（MB）杂交形成 DNA 双链，导致 MB 处于打开状态并产生荧光信号。随后，在等温循环扩增策略的进一步辅助下，设计的适配体传感平台显示出荧光信号的增加。由于这种放大策略的优势，检测限（LOD）降低至 0.74ng/mL，明显低于之前的报道。该策略不仅为定量分析低浓度目标蛋白提供了一个新的简单、特异、高效的平台，而且还展示了一种无需多次洗涤步骤的强大方法，具有将其集成到用于诊断应用的便携式、低成本和简化设备中的潜力。
