【摘要】：目前,對(duì)特異性DNA序列的檢測(cè)在疾病診斷中越顯重要,因此建立靈敏度高、特異性好的檢測(cè)特定DNA的方法就具有重要意義。其中,具有簡(jiǎn)單快速、低成本和高靈敏度的電化學(xué)檢測(cè)方法受到科學(xué)工作者的廣泛關(guān)注。在電化學(xué)檢測(cè)DNA的方法中,科學(xué)工作者設(shè)計(jì)的一種發(fā)夾型DNA因其本身具有優(yōu)異的特異性被應(yīng)用于電化學(xué)法對(duì)特異性DNA序列的檢測(cè)中。本論文的研究目的在于以發(fā)卡型DNA為探針,蘆丁為電活性物質(zhì),構(gòu)建一種基于空間構(gòu)象變化的、簡(jiǎn)單的發(fā)卡型DNA電化學(xué)生物傳感新方法,用于檢測(cè)不同目標(biāo)序列的DNA。第一章,首先介紹了DNA傳感器、DNA電化學(xué)生物傳感器和發(fā)卡型DNA電化學(xué)生物傳感器的原理、分類(lèi),對(duì)三種傳感器檢測(cè)特定序列DNA的方法分別進(jìn)行了闡述,并介紹了在各自領(lǐng)域的應(yīng)用;介紹了修飾于傳感器之上的金納米材料的應(yīng)用,最后闡述了本論文的研究目的和思路。第二章,構(gòu)建了一種性能優(yōu)良的基于發(fā)卡型DNA的電化學(xué)生物傳感器,實(shí)現(xiàn)了對(duì)慢性粒細(xì)胞性白血病相關(guān)基因b3a2的檢測(cè)。實(shí)驗(yàn)將金納米粒子和發(fā)卡型DNA依次修飾于電極表面,隨后將該電極上的發(fā)卡型DNA與目標(biāo)DNA結(jié)合,之后將該電極置于含有蘆丁的溶液中,采用電化學(xué)方法將蘆丁修飾于上述電極表面,并考察修飾于電極表面的蘆丁的電化學(xué)信號(hào)。實(shí)驗(yàn)發(fā)現(xiàn),修飾有發(fā)卡型DNA的電極在與目標(biāo)DNA結(jié)合前后,蘆丁的電化學(xué)信號(hào)明顯增強(qiáng),增強(qiáng)作用可能基于發(fā)卡型DNA與目標(biāo)DNA結(jié)合后,使電極表面空間構(gòu)象發(fā)生變化,更多的蘆丁可以被固定于電極表面,從而引起電極表面電化學(xué)信號(hào)的改變�；诖宋覀儗�(shí)現(xiàn)了對(duì)特異性序列目標(biāo)DNA的檢測(cè),并通過(guò)實(shí)驗(yàn)證明構(gòu)建的發(fā)卡型DNA傳感器具有良好的特異性識(shí)別能力。第三章,本章在第二章實(shí)驗(yàn)的基礎(chǔ)上,將此傳感器用于對(duì)嗜肺軍團(tuán)菌的相關(guān)基因進(jìn)行檢測(cè),進(jìn)一步驗(yàn)證了上述新型傳感方法的可行性。而實(shí)驗(yàn)結(jié)果證實(shí)了構(gòu)建的發(fā)卡型DNA電化學(xué)生物傳感新方法具有一定的適用性,可用于建立多種檢測(cè)不同序列的目標(biāo)DNA的分析方法,這為發(fā)卡型DNA電化學(xué)生物傳感器的發(fā)展拓寬了思路。第四章,本章分別研究了在熒光物質(zhì)羅丹明B、原卟啉、熒光素存在條件下,蘆丁電化學(xué)行為的改變,并采用紫外可見(jiàn)分光光度法和熒光分光光度法對(duì)其進(jìn)行了檢測(cè)。該研究對(duì)本論文所涉及的基于蘆丁電化學(xué)活性而建立的發(fā)卡型DNA電化學(xué)生物傳感器的設(shè)計(jì)具有重要研究意義。
[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.
【學(xué)位授予單位】：新疆師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：O657.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 王一嫻;葉尊忠;斯城燕;應(yīng)義斌;;適配體生物傳感器在病原微生物檢測(cè)中的應(yīng)用[J];分析化學(xué);2012年04期

2 ;DNA Immobilization on Nano-Gold Modified Glassy Carbon Electrode[J];Chinese Chemical Letters;1999年02期

3 王欽南,江虎軍,諸嘉佑;中國(guó)的人類(lèi)基因組計(jì)劃[J];生物工程進(jìn)展;1993年03期

相關(guān)碩士學(xué)位論文 前1條

1 劉旭慧;基于DNA模擬酶信號(hào)放大電化學(xué)阻抗DNA生物傳感器的研究[D];陜西師范大學(xué);2014年

，

本文編號(hào)：2292895