Analytical Methods Based On Thrombin And Adenosine Aptamer Functionalized Nano-probe Recognition

Nano-probe technique and aptamer recognition are novel probe technique and moleculars recognition mode developing rapidly in recent years, which have presented tremendously potential in the application of biological analysis, drug testing, food safety detection, clinical verify, environmental analysis and the others. Luminescnect analysis with the merits of high sensitivity and wide linear range has also attracted more and more attentions recently. This paper means to construct novel nano-probes using thrombin and adenosine binding with their specific aptamer-functionalized Au nanoparticles, combining with chemiluminescence detecting technology, the Au and Ag staining signal amplification technology and resonance light scattering technology, establish methods systems suit for highly sensitive detection of thrombin and adenosine. In the meanwhile, electrochemi -luminescece analysis was combined with capillary electrophoresis separation to establish a rapidly and sensitively analytical method for clarithromycin determination in biological fluids. All these would be of importance for enriching the theory and accelerating the applications of nano-probe and luminescent analysis technique.Firstly, combining the aptamer-functionalized Au nanoparticles with aptamer recognition technology, using thrombin as the model analyte, a novel analytical method for thrombin was developed coupling with flow-injection nanogold stripping chemiluminescence analysis technique.The dectiong limit of this method is 0.3×10-10 g/mL.Secondly, based on above reseach, using thrombin as model analyte, introducing its specific aptamer recognise thrombin, thrombin-specific aptamer functionalized Au nano-probe were assembled to the solid matrix surface. Then a selective gold deposition procedure on gold nanoparticles surface was carried out to realize the first signal amplification. Stripping chemiluminescence detection was conducted to further amplify the detection signals, making the sensitivity of the biological sensing system enhanced greatly.Thirdly, considering the problem of time consumption in gold stain, similar to gold staining based stripping CL detection, Chapter IV selectively deposite silver on the surface of nanogold for the first in-situ signal amplification, and then bond with the stripping chemiluminescence detection to carry out the second signal amplification, established a novel ultra-trace detection method for thrombin. The dectiong limit of this method is 0.3×10-12 g/mL.Finally, coupling nano-probe technology, aptamer recognition with resonance light scattering analysis, through recognising reaction between adenosine and its specific aptamer in homogeneous matrix, causing the deaggregation of the aggregated network structure of adenosine specific aptamer functionalized nanogold structure, a sharply change on the resonance light scattering signal can be obtained. Based on it, a novel homogeneous aptamer recognizing based method for adenosine detection was established.