Based On Aptamer Chemiluminescence Detection Of Small Bio-molecules Like Adenosine

This paper report a detection of biological small molecule adenoisine whichbased on aptamer chemiluminescence(CL), the object of the research is adenosine,carboxyl modified magnetic microspheres as separate carrier. Aptamer connected tomagnetic microsphere through aminoxatyl reaction, then report the sequence throughbase-pairing principle. Due to the effect of competition of the adenosine, leading thedecrease of CL matter which connected to magnetic microspheres, causing theweaken of signal. Thus indirectly achieve the detection of adenosine. Theseexperiments construct three detection methods, which shows below:The first detection method is using a horseradish peroxidase chemiluminescencedetection system. The experiment optimize the amount of carboxyl-modifiedmagnetic microspheres, aptamers, and streptavidin-modified horseradish peroxidaseand also investigate the selective of the uridine, cytidine and guanosine to adenosineaptamer. The results shows good linear relationship between the concentration ofadenosine in the1×10-9mol/L~10-4mol/L as well as the good performance ofchemiluminescent signal, the minimum detectable concentration of adenosine reaches1pmol/L; In addiction, the actual urinary samples and recovery experiments show thatthe method is feasible.The second detection method use liquid bromine-gold nanoparticleschemiluminescence detection system and develop a new technology basd on aptamers-gold nanoparticles chemiluminescence detection of adenosine. The experimentsoptimize the amount of carboxyl-modified magnetic microspheres, aptamer reportsequence and gold nanoparticles labeled streptavidin-gold and detect the adenosine inactual urine after conducting selective experiments. In addition, the goldnanoparticles is prepared by gold chloride reducting sodium citrate acid, then producethe streptavidin-labeled gold nanoparticles by chelation reaction. Achieving indirectchemiluminescence detection of biological small molecule adenosine.The third method follows the previous step part of the study.It establish ahydroxylamine-gold chloride acid amplification system, which enhance the the catalytic ability of luminol chemiluminescence system significantly as well as thesensitivity. Likewise, It optimize the reaction conditions required and selectiveexperiments. While under optimal experimental conditions it achieve the detection ofadenosine, which reaches a high detection limit100fmol/L; It proves the detectonmethod has a good good selectivity, stability and repeatability by conducting specificand recovery experiments. Also, it provides a satisfactory results by detecting theactual adenosine in cancer patient’surine. Therefore, this simple operative and highsensitivity method is expected to provide a favorable clinical diagnosis of thedisease.