【作者】 彭晓丽；

【导师】 乐国伟；

【作者基本信息】 江南大学 ， 营养与食品卫生学， 2008， 硕士

【摘要】 纳米探针技术和核酸适配体识别是近年来迅速发展的新型探针技术和分子识别模式,在生物分析、药物检测、食品安全检测、临床检验、环境分析等方面表现出了巨大的应用潜力。发光分析技术因为具有灵敏度高、线性范围宽等优点,近年来也越来越受重视。论文以凝血酶和腺苷特异结合核酸适配体功能化金纳米粒子构建新型纳米探针,结合化学发光检测技术和金染、银染信号放大技术,以及均相共振光散射技术,建立了适用于凝血酶和腺苷高灵敏检测的方法体系。相关分析技术的发展,对于丰富纳米探针和发光分析技术理论,拓宽其应用具有积极意义。首先,将核酸适配体功能化金纳米粒子与核酸适配体识别技术相结合,以凝血酶为模式分析物,结合纳米金溶出流动注射化学发光分析技术,建立基于核酸适配体功能化金纳米粒子识别的凝血酶化学发光新型检测方法。方法检测限达0.3×10-10 g/mL。其次,在前面研究的基础上,以凝血酶蛋白为模式分析物,采用凝血酶特异核酸适配体识别凝血酶,并通过特异识别反应将凝血酶核酸适配体功能化纳米金探针组装到固相表面,进一步在纳米金颗粒表面进行金的选择性沉积,实现第一次信号放大,然后采用溶出化学发光检测实现第二次信号放大,使得该生物传感系统的灵敏度大大提高。接着,针对金染化学溶出相对费时的问题,第四章继续以凝血酶蛋白为模式分析物,采用凝血酶特异核酸适配体识别凝血酶,并通过特异识别反应将凝血酶核酸适配体功能化纳米金探针组装到固相表面,进一步在纳米金颗粒表面进行银的选择性沉积,实现第一次原位信号放大,然后采用溶出化学发光检测实现第二次信号放大,建立一种适用于凝血酶超痕量检测的新型方法体系。方法检测限达0.3×10-12 g/mL。最后,将纳米探针技术、核酸适配体识别与共振光散射检测技术联用,通过均相体系中腺苷与其特异结合核酸适配体识别反应,使形成网络聚集结构的金纳米结构解聚,可引起共振光散射信号强烈变化。基于此,建立了核酸适配体识别的均相检测腺苷的新方法。更多还原

【Abstract】 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.更多还原