本文關(guān)鍵詞： 電化學(xué)傳感 核酸適體 雙信號(hào) 疾病標(biāo)志物 溶菌酶　出處：《青島大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：疾病標(biāo)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(cè)以及藥物篩選等的重要依據(jù)。然而大多數(shù)的疾病標(biāo)志物在人體中含量極低,因此很難對(duì)實(shí)際樣品中疾病標(biāo)志物準(zhǔn)確檢測(cè),這種情況極大地制約了疾病標(biāo)志物檢測(cè)在臨床實(shí)踐中的應(yīng)用。電化學(xué)核酸適體傳感器(E-AB)因具有了高的靈敏度、強(qiáng)的特異性、快的響應(yīng)速度、易于微型化和集成化等特點(diǎn)被廣泛應(yīng)用于分析檢驗(yàn)中。本文以檢測(cè)實(shí)際樣品中超低濃度疾病標(biāo)志物為目的,通過設(shè)計(jì)不同的雙信號(hào)策略建立了兩種雙信號(hào)E-AB以實(shí)現(xiàn)對(duì)溶菌酶的便捷、快速、精準(zhǔn)檢測(cè)。這對(duì)于一些重大疾病的病前預(yù)防、早期診斷及治療監(jiān)測(cè)等方面具有十分重要的意義。本研究設(shè)計(jì)了一種結(jié)合“signal-on/signal-off"和“l(fā)abeling/label-free”的策略構(gòu)建的E-AB用于對(duì)溶菌酶的高靈敏與選擇性檢測(cè)。首先,將二茂鐵修飾在單鏈DNA上作為一個(gè)電化學(xué)信號(hào)標(biāo)記物以提供“signal-on”的信號(hào);其次,帶正電的六氨合釕離子通過靜電作用吸附在帶負(fù)電荷的DNA磷酸骨架上作為另一個(gè)電化學(xué)信號(hào)標(biāo)記物以提供免標(biāo)記的“signal-off'的信號(hào)。與單信號(hào)相比較,雙信號(hào)的疊加(|AIRuHex| +△IFc)提高了該傳感器對(duì)溶菌酶檢測(cè)的靈敏度,檢測(cè)限達(dá)到0.8 pM�；趯⒔饘偌{米顆粒的比色檢測(cè)方法“嫁接”到電化學(xué)檢測(cè)平臺(tái)上的策略,本研究設(shè)計(jì)了一種可以用于檢測(cè)溶菌酶的傳感方法。為了實(shí)現(xiàn)該“嫁接”過程,我們首先制備了一種基于目標(biāo)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(cè)原理“嫁接”到電化學(xué)平臺(tái)上制備出操作簡(jiǎn)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(cè)限:0.9nM)相比,制得的E-AB線性范圍更寬(1pM-10nM),檢測(cè)限更低,可達(dá)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(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R446;O657.1;TP212

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