基于邻位诱导杂交信号开关的非标记拉曼光谱检测核酸和蛋白质新方法的研究

Nucleic acid and protein assay has already taken a key place in molecular diagnosis, forensic investigations etc. Ultrasensitive detection method for nucleotide and protein could realize the determination at exceptionally low level, of which the label signal amplification protocol prevailed. However, the fussy labeling process, analysis of high cost, and might result in the deactivation of DNA held it back. In view of the unique Raman characteristic peaks of adenine bases at 736 cm-1, this project proposes a novel label free Raman biosensors based on the efficiently surface Raman enhancement capability of silver metal nanoparticles, adenine bases composed of DNA as a signal probe, and proximity hybridization regulated biogate DNA assembly new technology. Various spectroscopic and morphological characterizations are applied for real-time monitoring the silver metal nanoparticles growth on the substrate to reveal the reduction effect of different reductants, and nonspecific surface adsorption etc. conditions for system optimization, and proximity hybridization regulated DNA assembly technology and adenine as internal signal mechanism for further study. Furthermore, the optimization of preparation and detection parameters would promise a nice sensing performance. The proposed method will be practically utilized to detect specific cancer-related expressing genes and markers, which would not only lay a foundation for clinic diagnoses.

核酸和蛋白质分析已在分子诊断、法医鉴定等领域发挥了重要作用。高灵敏核酸和蛋白质检测方法可实现超低浓度的分析，这其中以标记信号放大技术尤为突出。但是该技术目前因标记分析成本高、合成步骤繁琐，以及影响DNA的活性而受限制。鉴于DNA中的腺嘌呤碱基在736 cm-1处独特的拉曼特征峰，本项目提出，利用银金属纳米颗粒高效的表面拉曼增强能力，以腺嘌呤碱基组成的DNA作为信号探针，并联用邻位诱导杂交信号开关的DNA组装新技术，研制新型的多功能非标记拉曼生物传感器。并应用多种先进表征手段探索银纳米颗粒在基底上的生长形貌，揭示不同还原剂的还原效果，并且对表面非特异性吸附控制等条件进行系统的优化，对邻位诱导策略DNA组装技术方式和腺嘌呤作为内在信号机理进行深入的探究，继而优化参数以获得最佳分析性能。最终将为重大疾病如癌症基因和标志物的分析提供新方法和新技术，为临床诊断奠定基础。

核酸和蛋白质分析已在分子诊断、法医鉴定等领域发挥了重要作用。高灵敏核酸和蛋白质检测方法可实现超低浓度的分析。利用银金属纳米颗粒高效的表面拉曼增强能力，以腺嘌呤碱基组成的DNA作为信号探针，并联用邻位诱导杂交信号开关的DNA组装新技术，研制新型的多功能非标记拉曼生物传感器。探索银纳米颗粒在基底上的生长形貌，揭示不同还原剂的还原效果，并且对表面非特异性吸附控制等条件进行系统的优化，对邻位诱导策略DNA组装技术方式和腺嘌呤作为内在信号机理进行深入的探究，继而优化参数以获得最佳分析性能。. 构建若干种DNA调控生长的贵金属纳米颗粒，为生物分析领域提供新技术和新方法，获得较完善的高质量规则有序的贵金属纳米信号放大探针的合成工艺；制备若干种电活性高、稳定性好的DNA 调控生长的贵金属纳米探针，结合DNA 生物放大技术，并据此发展多种新颖的超灵敏电化学DNA 分析策略，并将该策略应用于实际体系，完成DNA 调控生长的贵金属纳米探针的性能分析和客观评价。为临床应用打下基础。