【摘要】：近年來,食品衛(wèi)生與安全及毒素的檢測成為備受關(guān)注的熱門話題,目前針對食品真菌毒素檢測,迫切需要高靈敏的檢測技術(shù),從而保障消費安全。傳統(tǒng)的生物傳感技術(shù)一般需要繁瑣的標(biāo)記過程,且靈敏度較差,本論文針對這些問題,結(jié)合非標(biāo)記均相傳感器的優(yōu)點以及能與目標(biāo)靶分子特異性結(jié)合的核酸適配體的優(yōu)勢發(fā)展出了兩種靈敏度高、選擇性好的新方法用于檢測食品中毒素。主要內(nèi)容如下:(1)提出了一種非標(biāo)記均相電化學(xué)方法用于來檢測黃曲霉毒素B1。該方法旨在以ITO電極表面長鏈DNA和短鏈DNA擴散性質(zhì)的不同和核酸外切酶的性質(zhì)相結(jié)合用于信號轉(zhuǎn)換系統(tǒng)。當(dāng)黃曲霉毒素B1存在時,其與核酸適配體反應(yīng)釋放的單鏈被外切酶切割擴散到電極表面,能觀察到明顯的電化學(xué)信號。(2)利用二茂鐵和亞甲基藍為信號指示劑的“信號開啟”和“信號關(guān)閉”體系,發(fā)展了一種新型的在均相溶液中反應(yīng)的可檢測多種毒素的電化學(xué)適配體生物傳感器。該方法設(shè)計了兩種與毒素有關(guān)的單鏈DNA通過金硫鍵自組裝固定到金電極表面,當(dāng)在均相溶液中加入兩種毒素時,這種雙鏈結(jié)構(gòu)就會解離,并通過形成毒素與適配體復(fù)合物釋放出信號指示劑標(biāo)記的DNA,來產(chǎn)生電化學(xué)信號。該方法的選擇性好,靈敏度高,成本低。其檢測OTA和AFB1的檢測下限分別為:0.04ng/mL、0.01ng/mL。
[Abstract]:In recent years, the detection of food hygiene and safety and toxins has become a hot topic of much concern. At present, high sensitive detection technology is urgently needed for the detection of food fungal toxins, so as to ensure the safety of consumption. Traditional biosensor technology usually requires tedious labeling process, and its sensitivity is poor. In this paper, we aim at these problems. Combining the advantages of heterogeneous sensors and the advantages of aptamers that can specifically bind to target molecules, two new sensitive and selective methods have been developed for the detection of toxins in food. The main contents are as follows: (1) A heterogeneous electrochemical method for the detection of aflatoxin B _ 1 was proposed. The purpose of this method is to combine the diffusion properties of long chain DNA and short chain DNA on the surface of ITO electrode and the properties of nucleic acid exonuclease to signal conversion system. When aflatoxin B1 exists, the single strand released by its reaction with aptamer of nucleic acid is cut by exonuclease and diffused to the electrode surface. Obvious electrochemical signals can be observed. (2) "signal on" and "signal off" systems using ferrocene and methylene blue as signal indicators, A novel electrochemical aptamer biosensor for the detection of multiple toxins in homogeneous solution was developed. In this method, two single-stranded DNA associated with toxins were designed to be self-assembled by gold-sulfur bond to the surface of the gold electrode. When two toxins were added into the homogeneous solution, the double-stranded structure would be dissociated. The electrochemical signal was generated by the formation of the toxin and aptamer complex and the release of signal indicator labeled DNA,. The method has good selectivity, high sensitivity and low cost. The detection limits of OTA and AFB1 were 0.04ng / mL and 0.01ng / mL, respectively.
【學(xué)位授予單位】：溫州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：TP212.3;TS207.5

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