本文選題：電化學DNA生物傳感　切入點：納米復合物　出處：《合肥工業(yè)大學》2017年碩士論文　論文類型：學位論文

【摘要】：近年來,隨著轉基因食品的出現(xiàn),轉基因食品的安全問題成為公眾擔心的問題。轉基因食品特定堿基序列的檢測越來越重要。開發(fā)簡便、迅速、廉價、微型化的DNA傳感器成為了研究熱點。電化學DNA傳感器因其響應較快選擇性好靈敏度高成本低等優(yōu)點,成為了生物學、醫(yī)學領域的新型課題。本文以特定轉基因堿基序列為檢測對象,利用血紅素-還原氧化石墨烯(Hemin-rGO)納米復合物、金納米粒子(Au NPs)、多壁碳納米管(WMCNTs)及硫堇-金納米粒子(THi-Au NPs)等納米材料修飾玻碳電極(GCE),構建了操作方便、靈敏度高、檢測限低的新型電化學DNA生物傳感器。對食品中轉基因成分進行分析。本文研究內容主要包含以下四個部分:1.Hemin-rGO納米復合物的制備及表征用氧化石墨及Hemin混合,然后加入還原劑水合肼及氨水,攪拌、油浴、離心清洗得到穩(wěn)定存在的Hemin-rGO納米復合物。用紫外-可見光光譜儀(UV-vis)和X射線光電子能譜分析(XPS)表征其結構及表面形態(tài)。用AuNPs和Hemin-rGO納米復合物修飾GCE制備電化學傳感器,并用電化學方法表征其電化學性能。2.基于Hemin-rGO納米復合物和Au NPs的玻碳電極超低檢測cDNA的無標記電化學基因傳感器用Au NPs和Hemin-rGO納米復合物修飾GCE制備電化學傳感器。由HS-DNA與cDNA堿基互補配對成為雙鏈DNA,阻斷Hemin的氧化還原反應,使電流相應降低。在沒有任何外部標簽下,構建一個簡單,靈敏和選擇性好的傳感平臺檢測cDNA。cDNA的濃度檢測范圍從1×10-18~1×10-13 M,系數(shù)為0.998,檢出限達到1.41×10-19 M。該方法對樣品施加和標準樣品的回收百分比進行了驗證。實驗表明該傳感器具有可靠性、高選擇性、再生性等優(yōu)點,是檢測DNA的理想平臺。3.THi-AuNPs-DNA納米復合物的制備與表征將THi固定到由氯金酸還原的Au NPs表面,生成THi-Au NPs納米復合材料。然后與經(jīng)活化的巰基標記DNA(S1)混合,成功制備了THi-Au NPs-S1納米復合材料。用紫外-可見光光譜儀對其結構及表面形態(tài)進行表征。并用電化學方法表征其電化學性能。4.基于氨基化的WMCNTs、THi-Au NPs-DNA納米復合物及辣根過氧化物酶雙重信號放大檢測DNA技術基于氨基化的WMCNTs、Au NPs、THi及辣根過氧化物酶修飾GCE制備電化學DNA傳感器。用辣根過氧化物酶催化過氧化氫的CV還原峰峰高來檢測DNA,達到雙重信號放大檢測DNA的目的。該電化學DNA傳感器的檢出限是3.524×10-21 M,可在適當?shù)臋z測條件下實現(xiàn)對轉基因食品的DNA的檢測。而且該電化學DNA傳感器具有良好的選擇性、穩(wěn)定性、重現(xiàn)性和高的再生性。
[Abstract]:In recent years, with the emergence of genetically modified foods, the safety of genetically modified foods has become a public concern. The detection of specific base sequences of genetically modified foods is becoming more and more important. Miniaturized DNA sensors have become a research hotspot. Electrochemical DNA sensors have a fast response. Good selectivity? High sensitivity? Because of its low cost, it has become a new subject in biology and medicine. In this paper, we used heme-reduced graphene oxide (Hemin-rGOA) nanocomposites to detect specific transgenic base sequences. The glassy carbon electrode modified with gold nanoparticles, such as au NPsO, WMCNTsand THi-Au NPs), has been constructed for easy operation and high sensitivity. A novel electrochemical DNA biosensor with low detection limit was used to analyze the genetically modified components in food. The main contents of this paper are as follows: 1. The preparation and characterization of Hemin-rgo nanocomposites are mixed with graphite oxide and Hemin. Then add the reducing agent hydrazine hydrate and ammonia water, stir, oil bath, The structure and surface morphology of Hemin-rGO nanocomposites were characterized by UV-vis spectrometer and X-ray photoelectron spectroscopy (XPS). Electrochemical sensors were prepared by modifying GCE with AuNPs and Hemin-rGO nanocomposites. Electrochemical properties were characterized by electrochemical methods .2. Electrochemical sensors modified with au NPs and Hemin-rGO nanocomposites modified with au NPs and Hemin-rGO nanocomposites were prepared by glassy carbon electrode with ultra-low detection of cDNA based on Hemin-rGO nanocomposites and au NPs. HS-DNA and cDNA bases were paired to double strand DNA to block the redox reaction of Hemin. Reduce the current accordingly. Without any external label, build a simple, The detection range of cDNA.cDNA concentration on a sensitive and selective sensor platform is from 1 脳 10 ~ (-18) to 1 脳 10 ~ (-13) M, the coefficient is 0.998, and the detection limit is 1.41 脳 10 ~ (-19) M. this method is used to verify the application of the sample and the recovery percentage of the standard sample. The experimental results show that the sensor is reliable. High selectivity and reproducibility are the ideal platform for detecting DNA. 3. Preparation and characterization of THi-AuNPs-DNA nanocomposites. THi is immobilized on the surface of AuNPs reduced by chloruronic acid to form THi-AuNPs nanocomposites, and then mixed with activated mercapto labeled DNAS1. THi-Au NPs-S1 nanocomposites were successfully prepared. Their structures and surface morphology were characterized by UV-Vis spectrometer. The electrochemical properties of the nanocomposites were characterized by electrochemical method. 4. WMCNTsTsTHi-Au NPs-DNA nanocomposites and horseradish compounds based on amination. Peroxidase double signal Amplification Detection (DNA) technique based on Aminalized WMCNTsN au NPsNS-THi and horseradish peroxidase modified GCE to prepare electrochemical DNA sensor, GCE was used to detect the peak height of CV reduction peak of hydrogen peroxide catalyzed by horseradish peroxidase. The detection limit of the electrochemical DNA sensor is 3.524 脳 10 ~ (-21) M.It can be used to detect the DNA of transgenic food under appropriate detection conditions, and the electrochemical DNA sensor has good selectivity. Stability, reproducibility and high reproducibility.
【學位授予單位】：合肥工業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：TS207.3;O657.1

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本文編號：1557878