基于核酸侧向层析的microRNA快速检测新方法建立

发布时间：2018-01-04 01:38

本文关键词：基于核酸侧向层析的microRNA快速检测新方法建立　出处：《合肥工业大学》2017年硕士论文　论文类型：学位论文

【摘要】：随着人们生活节奏的加快和饮食结构的变化,罹患癌症的人群增多,对癌症的前期诊断和术后的监控尤为重要。而MicroRNA的表达水平和癌症的发生紧密相连。此外,microRNA可通过日常食物摄取进入人体内,从而调控人体靶基因表达方式,进而影响人体的生理功能。MicroRNA种类繁多,因此建立microRNA的快速、简单、高灵敏的多重检测技术在医学、食品营养与安全等领域具有重要意义。本文建立了以下两种检测方法:(1)基于侧向层析核酸试纸条多重检测microRNA本方法利用三组Au-SH probe-T probe“三明治”夹心结构的形成实现对microRNA的三重检测。在金纳米粒子表面固定检测探针SH probe,NC膜上固定捕获探针T probe,目标物microRNA可与SH probe和T Probe部分杂交,形成夹心结构从而将金纳米粒子捕获并滞留在NC膜上,通过金纳米粒子在检测线上的累积从而出现肉眼可见的条带。建立的单重核酸试纸条对microRNA-21,microRNA-155和microRNA-210肉眼检测限分别为1 nM,5 nM和5 nM;建立的多重核酸试纸条可以很好地实现对三种目标物的同时检测且不发生交叉反应。(2)基于侧向层析技术的表面增强拉曼光谱双重检测microRNA本方法是建立在Au@Ag-SH probe-T probe夹心结构形成的基础上,在Au@Ag上固定检测探针之前先在其表面修饰拉曼信号分子。当修饰了拉曼信号分子的Au@Ag被滞留在检测线上时,通过肉眼根据检测线的强度可以初步判断待测物浓度,而通过拉曼光谱分析,在肉眼无法分辨的浓度范围内也能检出信号,从而提高检测灵敏度,对microRNA-21和microRNA-155的拉曼检测灵敏度可达到0.5 nM和1 nM。此外,根据Rho-6G和4-ATP拉曼特征峰的不同实现了在试纸条上单线检测microRNA-21和microRNA-155。
[Abstract]:With the accelerated pace of life and changes in diet, cancer population increased early diagnosis and monitoring of cancer after surgery is particularly important. And the expression level of MicroRNA and cancer are closely linked. In addition, microRNA through the daily food intake into the body, the body to regulate the expression of target gene, and then physiological function of human.MicroRNA type is various, so the establishment of microRNA rapid, simple, high sensitive multi detection technology has important significance in medicine, nutrition and food security and other fields. This paper established the following two methods: (1) lateral chromatography nucleic acid strip multiple detection method using the microRNA Au-SH probe-T group formed three probe "sandwich" structure to achieve three detection based on microRNA. The gold nanoparticle surface fixed detection probe SH probe NC immobilized on the membrane The capture probe T probe, microRNA and SH can target probe and T Probe hybrid, which will form a sandwich structure of gold nanoparticles capture and retention in the NC membrane, the gold nanoparticles accumulated in the detection line to visible bands. A single nucleic acid test strip of microRNA-21, microRNA-155 and microRNA-210 eye detection limits were 1 nM, 5 nM and 5 nM; the establishment of the multiplex nucleic acid test strip can well realize three kinds of target detection at the same time and does not cross react. (2) surface enhanced Raman spectra of lateral tomography dual detection of microRNA this method is based on the formation of Au@Ag-SH probe-T probe sandwich structure based on Au@Ag before the fixed detection probe first coated on the surface of the Raman signal molecules. When modified Raman signal molecules of Au@Ag were stranded on the test line, through the eyes of flesh According to the detection of line intensity can determine the initial analyte concentration, and the Raman spectra analysis, signal was detected in the concentration range of the naked eye can not distinguish, thus improving the detection sensitivity of microRNA-21 and microRNA-155 Raman detection sensitivity can reach 0.5 nM and 1 nM. in addition, according to the Rho-6G and 4-ATP Raman peaks of different implementation in the test paper on the single detection of microRNA-21 and microRNA-155.

【学位授予单位】：合肥工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q78;O658.1

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