Jiangsu Engineering Laboratory of Smart Carbon-Rich Materials and Device, Jiangsu Province Hi-Tech Key Laboratory for Bio-medical Research, School of Chemistry and Chemical Engineering, Southeast University , Nanjing, 211189, China.
College of Food Science and Technology, Henan University of Technology , Zhengzhou, 450001, China.
Anal Chem. 2017 Nov 21;89(22):12094-12100. doi: 10.1021/acs.analchem.7b02608. Epub 2017 Nov 3.
Early diagnosis and life-long surveillance are clinically important to improve the long-term survival of cancer patients. Telomerase activity is a valuable biomarker for cancer diagnosis, but its measurement often used complex label procedures. Herein, we designed a novel, simple, visual and label-free method for telomerase detection by using enzymatic etching of gold nanorods (GNRs). First, repeating (TTAGGG) sequences were extented on telomerase substrate (TS) primer. It formed G-quadruplex under the help of Hemin and K. Second, the obtained horseradish peroxidase mimicking hemin/G-quadruplex catalyzed the HO-mediated etching of GNRs to the short GNRs, even to gold nanoparticles (GNPs), generating a series of distinct color changes due to their plasmon-related optical response. Thus, this enzymatic reaction can be easily coupled to telomerase activity, allowing for the detection of telomerase activity based on vivid colors. This can be differentiated sensitively by naked eyes because human eyes are more sensitive to color variations rather than the optical density variations. As a result, telomerase activity can be quantitatively detected ranging from 200 to 15000 HeLa cells mL. The detection limit was 90 HeLa cells mL (S/N = 3). Importantly, the application of this method in bladder cancer samples was in agreement with the clinical results. Thus, this method was considerably suitable for point-of-care diagnostics in resource-constrained regions because of the easy readout of results without the use of sophisticated apparatus.
早期诊断和终身监测对于提高癌症患者的长期生存率具有重要的临床意义。端粒酶活性是癌症诊断的有价值的生物标志物,但它的测量通常需要复杂的标记程序。在这里,我们设计了一种新颖、简单、直观且无需标记的方法,通过使用金纳米棒(GNRs)的酶蚀刻来检测端粒酶。首先,重复(TTAGGG)序列在端粒酶底物(TS)引物上进行延伸。在血红素和 K 的帮助下,它形成 G-四链体。其次,获得的辣根过氧化物酶模拟血红素/G-四链体催化 HO 介导的 GNRs 的蚀刻,形成短的 GNRs,甚至形成金纳米颗粒(GNPs),由于其等离子体相关的光学响应,产生一系列明显的颜色变化。因此,这种酶促反应可以很容易地与端粒酶活性偶联,从而可以根据生动的颜色来检测端粒酶活性。由于人眼对颜色变化比光密度变化更敏感,因此可以通过肉眼敏感地区分。结果,端粒酶活性可以从 200 到 15000 HeLa 细胞/mL 范围内进行定量检测。检测限为 90 HeLa 细胞/mL(S/N = 3)。重要的是,该方法在膀胱癌样本中的应用与临床结果一致。因此,由于无需使用复杂的仪器即可轻松读取结果,因此该方法非常适合资源有限地区的即时诊断。