基于“裂开型”核酸适体高灵敏检测生物小分子的研究

发布时间：2018-03-21 14:13

本文选题：“裂开型”核酸适体　切入点：单壁碳纳米管　出处：《湖南大学》2010年硕士论文　论文类型：学位论文

【摘要】：生物小分子的分析检测对疾病的预测和医学的诊断都具有重要的意义,发展并完善生物小分子的检测方法,使其更加快速、灵敏、准确和方便,是目前研究的热点之一。核酸适体由于具有易于合成和化学修饰、特异性好以及亲和力强等特点,且设计灵活,适用于多种信号转换方法,为生物小分子的分析检测提供了一种新的手段。本论文以生物小分子腺苷为检测对象,利用核酸适体能高特异性以及高亲和性识别小分子的特点,结合单壁碳纳米管和纳米金颗粒等多种纳米材料,通过不同的信号转换手段发展了基于“裂开型”核酸适体的检测方法,实现对生物小分子的简便、快速及灵敏的检测。具体如下： (1)基于“裂开型”核酸适体和单壁碳纳米管的生物小分子检测研究基于单壁碳纳米管-DNA复合物在高盐条件下的稳定性变化情况,利用单壁碳纳米管的类过氧化物酶活性,发展了一种生物小分子的检测方法。该方法无需标记,只需通过目测比色即可实现检测,检测下限为44 nM,灵敏度较高,特异性好,拓展了单壁碳纳米管-核酸适体体系的应用范围,并有望实现对其他小分子的检测。 (2)基于“裂开型”核酸适体和纳米金颗粒光散射信号的生物小分子检测研究基于“裂开型”核酸适体修饰的纳米金颗粒与腺苷特异性结合所引起的尺寸变化,利用纳米金颗粒的光散射信号,发展了一种生物小分子的检测方法。该方法仅需一步混合即可实现检测,简便、快速,检测下限为7 nM,灵敏度高,特异性好,有望发展简便、快速及灵敏的识别生物分子的方法。 (3)基于“裂开型”核酸适体和纳米金颗粒增强信号的表面等离子体共振传感器对生物小分子的检测基于“裂开型”核酸适体修饰的纳米金颗粒与腺苷特异性结合后将纳米金颗粒捕获到传感芯片表面,利用纳米金颗粒与金膜的电子耦合效应增强表面等离子体共振(SPR)信号,构建了直接检测生物小分子的高灵敏的SPR传感器件。该传感器灵敏度高,检测下限可达到1.5 pM,特异性好,芯片可再生,并且操作简便,拓展了表面等离子体共振传感技术的应用,并有望实现其他小分子的高灵敏检测。
[Abstract]:The analysis and detection of biological small molecules is of great significance to disease prediction and medical diagnosis. The development and improvement of biological small molecule detection methods to make it faster, more sensitive, accurate and convenient, Nucleic acid aptamers have the characteristics of easy synthesis and chemical modification, good specificity and strong affinity, and are flexible in design, and can be used in many signal conversion methods. In this paper, adenosine, a biological small molecule, is used to detect small molecules, which is characterized by high specificity and affinity of nucleic acid. Combined with single-walled carbon nanotubes and gold nanoparticles, the detection method based on "split" aptamer of nucleic acid was developed by different signal conversion methods, which is simple and convenient for biological small molecules. Rapid and sensitive detection. Details are as follows:. Detection of small biomolecules based on "split" aptamers and single-walled carbon nanotubes. Based on the stability changes of single-walled carbon nanotube-DNA complexes under high salt conditions, a new method for the detection of biomolecules was developed using the peroxidase activity of single-walled carbon nanotubes. The detection can be realized only by visual colorimetry, the detection limit is 44 nm, the sensitivity is high, the specificity is good, the application scope of single-walled carbon nanotube-nucleic acid aptamer system is expanded, and the detection of other small molecules is expected to be realized. Detection of small Biological molecules based on "split" Nucleic Acid aptamers and Light scattering signals from Gold nanoparticles. Based on the size change caused by the specific binding of the "split" aptamer modified gold nanoparticles with adenosine, the light scattering signals of the gold nanoparticles were used. A method for the detection of biological small molecules has been developed. The method can be detected with only one step mixing. The method is simple, rapid, and the detection limit is 7 nm, which has the advantages of high sensitivity, good specificity, and is expected to be easy to develop. A rapid and sensitive method for identifying biomolecules. Detection of small biomolecules by Surface Plasmon Resonance Sensor based on "split" Nucleic Acid aptamer and Nano-Gold Particle Enhancement signal. Based on the specific binding of the "split" aptamer modified gold nanoparticles to adenosine, the gold nanoparticles were captured on the surface of the sensor chip, and the surface plasmon resonance (SPRR) signal was enhanced by the electron coupling effect between the gold nanoparticles and the gold film. A highly sensitive SPR sensor for direct detection of biological small molecules was constructed. The sensor has high sensitivity, detection limit of 1.5 pm, good specificity, reproducible chip, and simple operation. The application of surface plasmon resonance sensing technology is expanded, and the high sensitivity detection of other small molecules is expected to be realized.
【学位授予单位】：湖南大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R341

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