Department of Biological Science and Technology, National Chiao Tung University, No. 75 Po-Ai Street, Hsinchu 300, Taiwan.
Department of Biological Science and Technology, National Chiao Tung University, No. 75 Po-Ai Street, Hsinchu 300, Taiwan.
Biosens Bioelectron. 2014 Nov 15;61:336-43. doi: 10.1016/j.bios.2014.05.007. Epub 2014 May 10.
Thrombin plays the role in cardiovascular diseases and regulates many processes in inflammation and could be a feature of many pathological conditions, including the thromboembolic disease, cancer and neurodegenerative diseases. An ultrasensitive and amplified electrochemical sandwich assay using screen-printed carbon electrode (SPCE) strips for thrombin detection was established in this study. The conductivity and sensing performance of the carbon electrodes were enhanced by using gold nanoparticles (AuNPs). The aptamer addressed on the strips was used as a primary probe to capture thrombin in the detected samples. An amplifier was invented for recognizing thrombin captured on the SPCE, which is the multiple molecules of anti-thrombin antibody (Ab) and horseradish peroxidase (HRP) co-modified AuNPs (AuNPs/Ab-HRP). Hydrogen peroxide was used as the substrate for HRP and then the response current (RC) could be detected. The optimization of these AuNPs conjugates-amplified aptamer immunosensing SPCE strips was conducted for thrombin detection. The detection sensitivity showed a linear relation between RC and thrombin concentration in the range of 10 pM-100 nM, and limit of detection (LOD) was 1.5 pM. The fabricated AuNPs/Ab-HRP-amplified aptamer immunosensing SPCE strips were further used to detect thrombin in human serum with a linear range of 100 pM-100 nM. This study provided the promising SPCE strips with highly sensitive and rapid detection for thrombin by the electrochemical aptasensor combined with AuNPs conjugates for amplifying the detection signal.
凝血酶在心血管疾病中起作用，调节炎症中的许多过程，可能是许多病理状况的特征，包括血栓栓塞疾病、癌症和神经退行性疾病。本研究建立了一种使用丝网印刷碳电极（SPCE）条检测凝血酶的超灵敏和放大电化学三明治检测方法。通过使用金纳米粒子（AuNPs）提高了碳电极的导电性和传感性能。将针对条带的适体用作捕获检测样品中凝血酶的初级探针。发明了一种用于识别 SPCE 上捕获的凝血酶的放大器，该放大器是抗凝血酶抗体（Ab）和辣根过氧化物酶（HRP）的多个分子共修饰的 AuNPs（AuNPs/Ab-HRP）。将过氧化氢用作 HRP 的底物，然后可以检测到响应电流（RC）。对这些 AuNPs 缀合物放大适体免疫传感 SPCE 条带进行了优化，以检测凝血酶。检测灵敏度显示 RC 与 10 pM-100 nM 范围内凝血酶浓度之间呈线性关系，检测限（LOD）为 1.5 pM。所制备的 AuNPs/Ab-HRP 放大适体免疫传感 SPCE 条带进一步用于检测人血清中的凝血酶，线性范围为 100 pM-100 nM。该研究通过电化学适体传感器与 AuNPs 缀合物结合用于放大检测信号，为凝血酶提供了具有高灵敏度和快速检测的有前途的 SPCE 条带。
