Gold nanoparticles conjugates-amplified aptamer immunosensing screen-printed carbon electrode strips for thrombin detection

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. (C) 2014 Elsevier B.V. All rights reserved.