本文關鍵詞： 電化學傳感 核酸適體 雙信號 疾病標志物 溶菌酶　出處：《青島大學》2017年碩士論文　論文類型：學位論文

【摘要】：疾病標志物通常是指一類具有特異性的生化指示劑,其可以作為疾病診斷、疾病監(jiān)測以及藥物篩選等的重要依據。然而大多數的疾病標志物在人體中含量極低,因此很難對實際樣品中疾病標志物準確檢測,這種情況極大地制約了疾病標志物檢測在臨床實踐中的應用。電化學核酸適體傳感器(E-AB)因具有了高的靈敏度、強的特異性、快的響應速度、易于微型化和集成化等特點被廣泛應用于分析檢驗中。本文以檢測實際樣品中超低濃度疾病標志物為目的,通過設計不同的雙信號策略建立了兩種雙信號E-AB以實現對溶菌酶的便捷、快速、精準檢測。這對于一些重大疾病的病前預防、早期診斷及治療監(jiān)測等方面具有十分重要的意義。本研究設計了一種結合“signal-on/signal-off"和“l(fā)abeling/label-free”的策略構建的E-AB用于對溶菌酶的高靈敏與選擇性檢測。首先,將二茂鐵修飾在單鏈DNA上作為一個電化學信號標記物以提供“signal-on”的信號;其次,帶正電的六氨合釕離子通過靜電作用吸附在帶負電荷的DNA磷酸骨架上作為另一個電化學信號標記物以提供免標記的“signal-off'的信號。與單信號相比較,雙信號的疊加(|AIRuHex| +△IFc)提高了該傳感器對溶菌酶檢測的靈敏度,檢測限達到0.8 pM�；趯⒔饘偌{米顆粒的比色檢測方法“嫁接”到電化學檢測平臺上的策略,本研究設計了一種可以用于檢測溶菌酶的傳感方法。為了實現該“嫁接”過程,我們首先制備了一種基于目標物引發(fā)下DNA-金納米顆粒傳感單元聚集的比色核酸適體傳感器,之后將該檢測原理“嫁接”到電化學平臺上制備出操作簡便、靈敏度高和穩(wěn)定性好的E-AB。與比色核酸適體傳感器(線性范圍:5nM-500nM;檢測限:0.9nM)相比,制得的E-AB線性范圍更寬(1pM-10nM),檢測限更低,可達到0.3 pM。
[Abstract]:Disease markers usually refer to a class of specific biochemical indicators, which can be used as an important basis for disease diagnosis, disease surveillance and drug screening. However, most disease markers are extremely low in the human body. Therefore, it is difficult to detect disease markers accurately in real samples, which greatly restricts the application of disease markers in clinical practice. Electrochemical aptamer sensor E-ABM has high sensitivity and specificity. The characteristics of rapid response, easy miniaturization and integration are widely used in the analysis and testing. The purpose of this paper is to detect ultralow concentration disease markers in actual samples. By designing different dual signal strategies, two kinds of double signal E-AB are established to realize the convenient, rapid and accurate detection of lysozyme. Early diagnosis and treatment monitoring are of great significance. In this study, an E-AB strategy combined with "signal-on/signal-off" and "labeling/label-free" was designed to detect lysozyme with high sensitivity and selectivity. Ferrocene was modified on single-stranded DNA as an electrochemical signal marker to provide "signal-on" signal. The positively charged ruthenium hexaamine ion is adsorbed on the negatively charged DNA phosphoric acid skeleton by electrostatic interaction as another electrochemical signal marker to provide a signal-free "signal-off" signal, compared with a single signal. Double signal superposition (AIRuHex IFC) improves the sensitivity of the sensor to lysozyme detection, reaching a detection limit of 0.8 pM.Based on the strategy of "grafting" the metal nanoparticles colorimetric detection method to the electrochemical detection platform, In this study, we designed a sensing method for detecting lysozyme. In order to realize the grafting process, we first prepared a colorimetric nucleic acid aptamer sensor based on DNA-gold nanocrystalline sensor unit. The detection principle was then "grafted" onto the electrochemical platform to prepare E-ABs with simple operation, high sensitivity and good stability. Compared with colorimetric nucleic acid aptamer sensors (linear range: 5nM-500nM; detection limit: 0.9nMM), The linear range of E-AB is wider than that of 1pM-10nMN, and the detection limit is lower, reaching 0.3 pM.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R446;O657.1;TP212

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