连接酶链式反应快速灵敏的检测microRNA研究.pdf

河北大学 硕士学位论文 连接酶链式反应快速灵敏的检测microRNA 姓名：严景丽 申请学位级别：硕士 专业：分析化学 指导教师：李正平 Abstract Abstract MicroRNA (miRNA) play an important regulatory role in biological systems. Studies of the biological functions of miRNA are have great potential for early clinical diagnosis and diacovery of new drugs. Developing simple, rapid, highly sensitive and specific methods for miRNAs detection are very important for microRNA studies. In this dissertation, by using miRNAs as a template, DNA probes immobilized on different gold nanoparticles were successfully ligated, which resulted in the cross-linking aggregation of gold nanoparticles. Therefore miRNAs can be simply detected by visualization of the color changes. This method was simple and easy to operate, and does not need any method of amplification and expensive equipment. The results of this method can be distinguished with naked eyes, which facilitated its application. The limitation of the proposed method is that it is difficult to carry out the ligase chain reaction (LCR), so the sensitivity is not satisfactory. On this basis, we developed a simple and sensitive method, in which let-7a miRNA was used as the template and T4 RNA ligase 2 was used to selectively ligate DNA probes. Thus the LCR was initiated and the products were separated and detected by agarose gel electrophoresis. The detection limit (3σ, n = 7) of Let-7a was estimated to be 0.7 pmol/L. There is a good correlative relationship between the pixel intensi