An Electrochemical Biosensor for Detection of Thrombin based on Displacement Mode

摘要

A sensitive electrochemical biosensor for detection of thrombin based on target protein-induced strand displacement is presented. For this proposed biosensor, dsDNA which was prepared by the hybridization reaction of the immobilized probe ssDNA (IP) containing thiol group and thrombin aptamer base sequence was initially immobilized on the Au electrode by self-assemble via Au-S bind, and a DNA labeled with PdS nanoparticles (DP-PdS) was used as a detection probe. When the so prepared dsDNA modified Au electrode was immersed into a solution containing target protein and DP-PdS, the aptamer in the dsDNA give priority to to form G-quarter structure with the present target protein and the dsDNA sequence was released one single strand and returned to 1P strand which consequently hybridized with DP-PdS. After dissolving the captured PdS particles from the electrode, a mercury-film electrode was used for electrochemical detection of these Pd~(2+) ions which offered sensitive electrochemical signal transduction. The peak current of Pd~(2+) ions had a good linear relationship with the thrombin concentration in the range of 7.3 × 10~(-8)-7.3 × 10~(-11) mol/L and the detection limit was 2.3 × 10~(-11) mol/L of thrombin. The detection was also specific for thrombin without being affected by the coexistence of other proteins, such as BSA and lysozyme.