基于发夹结构DNA空间构象变化的电化学生物传感器的研究
发布时间：2018-10-25 06:28
【摘要】：目前,对特异性DNA序列的检测在疾病诊断中越显重要,因此建立灵敏度高、特异性好的检测特定DNA的方法就具有重要意义。其中,具有简单快速、低成本和高灵敏度的电化学检测方法受到科学工作者的广泛关注。在电化学检测DNA的方法中,科学工作者设计的一种发夹型DNA因其本身具有优异的特异性被应用于电化学法对特异性DNA序列的检测中。本论文的研究目的在于以发卡型DNA为探针,芦丁为电活性物质,构建一种基于空间构象变化的、简单的发卡型DNA电化学生物传感新方法,用于检测不同目标序列的DNA。第一章,首先介绍了DNA传感器、DNA电化学生物传感器和发卡型DNA电化学生物传感器的原理、分类,对三种传感器检测特定序列DNA的方法分别进行了阐述,并介绍了在各自领域的应用;介绍了修饰于传感器之上的金纳米材料的应用,最后阐述了本论文的研究目的和思路。第二章,构建了一种性能优良的基于发卡型DNA的电化学生物传感器,实现了对慢性粒细胞性白血病相关基因b3a2的检测。实验将金纳米粒子和发卡型DNA依次修饰于电极表面,随后将该电极上的发卡型DNA与目标DNA结合,之后将该电极置于含有芦丁的溶液中,采用电化学方法将芦丁修饰于上述电极表面,并考察修饰于电极表面的芦丁的电化学信号。实验发现,修饰有发卡型DNA的电极在与目标DNA结合前后,芦丁的电化学信号明显增强,增强作用可能基于发卡型DNA与目标DNA结合后,使电极表面空间构象发生变化,更多的芦丁可以被固定于电极表面,从而引起电极表面电化学信号的改变。基于此我们实现了对特异性序列目标DNA的检测,并通过实验证明构建的发卡型DNA传感器具有良好的特异性识别能力。第三章,本章在第二章实验的基础上,将此传感器用于对嗜肺军团菌的相关基因进行检测,进一步验证了上述新型传感方法的可行性。而实验结果证实了构建的发卡型DNA电化学生物传感新方法具有一定的适用性,可用于建立多种检测不同序列的目标DNA的分析方法,这为发卡型DNA电化学生物传感器的发展拓宽了思路。第四章,本章分别研究了在荧光物质罗丹明B、原卟啉、荧光素存在条件下,芦丁电化学行为的改变,并采用紫外可见分光光度法和荧光分光光度法对其进行了检测。该研究对本论文所涉及的基于芦丁电化学活性而建立的发卡型DNA电化学生物传感器的设计具有重要研究意义。
[Abstract]:At present, the detection of specific DNA sequences is more and more important in the diagnosis of disease. Therefore, it is of great significance to establish a sensitive and specific method for the detection of specific DNA. Among them, simple, fast, low cost and high sensitivity electrochemical detection methods have been widely concerned by scientists. In electrochemical detection of DNA, a hairpin type DNA designed by scientists has been applied to the detection of specific DNA sequences by electrochemical method because of its excellent specificity. The purpose of this thesis is to construct a new electrochemical biosensor for DNA based on spatial conformation change, using hairpin type DNA as probe and rutin as electroactive material, which can be used to detect DNA. of different target sequences. In the first chapter, the principle and classification of DNA sensor, DNA electrochemical biosensor and card type DNA electrochemical biosensor are introduced. The applications of gold nanomaterials modified on the sensor are introduced, and the purpose and thinking of this thesis are described. In chapter 2, an electrochemical biosensor based on card type DNA was constructed to detect the b3a2 of chronic myeloid leukemia (CML) related gene. The gold nanoparticles and hairpin type DNA were modified on the surface of the electrode in turn, then the hairpin type DNA on the electrode was bonded with the target DNA, and then the electrode was placed in the solution containing rutin. Rutin was modified on the surface of the electrode by electrochemical method, and the electrochemical signal of rutin modified on the electrode was investigated. It was found that the electrochemical signal of rutin was obviously enhanced before and after the electrode modified with hairpin type DNA combined with the target DNA. The enhancement effect may be based on the spatial conformation of the electrode surface after the combination of the hairpin type DNA and the target DNA. More rutin can be fixed on the electrode surface, resulting in the change of electrochemical signal on the electrode surface. Based on this, we have realized the detection of specific sequence target DNA, and the experimental results show that the card type DNA sensor has good specificity recognition ability. In chapter 3, based on the experiment in chapter 2, the sensor is used to detect the genes related to Legionella pneumophila, which further verifies the feasibility of the new sensing method. The experimental results show that the new DNA electrochemical biosensor method has some applicability and can be used to establish a variety of target DNA analysis methods for detecting different sequences. This opens the way for the development of card-type DNA electrochemical biosensor. In chapter 4, the electrochemical behavior of rutin in the presence of Rhodamine B, protoporphyrin and fluorescein was studied, and the changes of rutin electrochemical behavior were determined by UV-Vis spectrophotometry and fluorescence spectrophotometry. This study is of great significance to the design of hairpin type DNA electrochemical biosensor based on rutin electrochemical activity.
【学位授予单位】：新疆师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.1
本文编号：2292895
[Abstract]:At present, the detection of specific DNA sequences is more and more important in the diagnosis of disease. Therefore, it is of great significance to establish a sensitive and specific method for the detection of specific DNA. Among them, simple, fast, low cost and high sensitivity electrochemical detection methods have been widely concerned by scientists. In electrochemical detection of DNA, a hairpin type DNA designed by scientists has been applied to the detection of specific DNA sequences by electrochemical method because of its excellent specificity. The purpose of this thesis is to construct a new electrochemical biosensor for DNA based on spatial conformation change, using hairpin type DNA as probe and rutin as electroactive material, which can be used to detect DNA. of different target sequences. In the first chapter, the principle and classification of DNA sensor, DNA electrochemical biosensor and card type DNA electrochemical biosensor are introduced. The applications of gold nanomaterials modified on the sensor are introduced, and the purpose and thinking of this thesis are described. In chapter 2, an electrochemical biosensor based on card type DNA was constructed to detect the b3a2 of chronic myeloid leukemia (CML) related gene. The gold nanoparticles and hairpin type DNA were modified on the surface of the electrode in turn, then the hairpin type DNA on the electrode was bonded with the target DNA, and then the electrode was placed in the solution containing rutin. Rutin was modified on the surface of the electrode by electrochemical method, and the electrochemical signal of rutin modified on the electrode was investigated. It was found that the electrochemical signal of rutin was obviously enhanced before and after the electrode modified with hairpin type DNA combined with the target DNA. The enhancement effect may be based on the spatial conformation of the electrode surface after the combination of the hairpin type DNA and the target DNA. More rutin can be fixed on the electrode surface, resulting in the change of electrochemical signal on the electrode surface. Based on this, we have realized the detection of specific sequence target DNA, and the experimental results show that the card type DNA sensor has good specificity recognition ability. In chapter 3, based on the experiment in chapter 2, the sensor is used to detect the genes related to Legionella pneumophila, which further verifies the feasibility of the new sensing method. The experimental results show that the new DNA electrochemical biosensor method has some applicability and can be used to establish a variety of target DNA analysis methods for detecting different sequences. This opens the way for the development of card-type DNA electrochemical biosensor. In chapter 4, the electrochemical behavior of rutin in the presence of Rhodamine B, protoporphyrin and fluorescein was studied, and the changes of rutin electrochemical behavior were determined by UV-Vis spectrophotometry and fluorescence spectrophotometry. This study is of great significance to the design of hairpin type DNA electrochemical biosensor based on rutin electrochemical activity.
【学位授予单位】：新疆师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.1
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