本文關(guān)鍵詞： 熒光免疫分析 熒光共振能量轉(zhuǎn)移 ZnS Mn量子點(diǎn) CdTe量子點(diǎn) 金黃色葡萄球菌腸毒素B 快速檢測(cè)　出處：《湖南師范大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：金黃色葡萄球菌腸毒素B(SEB)是由一種革蘭氏陽性球菌產(chǎn)生的常見的食物毒素。作為一種超抗原,SEB在炎癥過程中起到非常重要的作用。它也是過敏性疾病中的一種重要的致敏原。由于其內(nèi)在的高穩(wěn)定性、強(qiáng)毒性和易散播的特性,SEB還是一種潛在的生物武器。所以發(fā)展一種快速、簡(jiǎn)便、靈敏的SEB檢測(cè)新方法具有非常重要的意義。 熒光免疫分析靈敏度高、特異性好,是一種應(yīng)用十分廣泛的生物、生化分析技術(shù)。鑒此,本論文在本實(shí)驗(yàn)室前期研究的基礎(chǔ)上,首先建立了一種以CdTe量子點(diǎn)作為能量供體、熒光染料Alexa Fluor568(AF568)作為能量受體的熒光共振能量轉(zhuǎn)移分析體系,用來快速檢測(cè)SEB毒素。但是考慮到Cd系列量子點(diǎn)的毒性,本論文進(jìn)一步建立了基于ZnS摻Mn低毒性量子點(diǎn)的熒光免疫分析,并將之用于毒素SEB的快速、靈敏檢測(cè),以應(yīng)對(duì)食品安全監(jiān)測(cè)的迫切需要。本論文工作主要內(nèi)容： 1.綜述了熒光免疫分析技術(shù)的應(yīng)用以及現(xiàn)階段常用于熒光分析中的不同熒光標(biāo)記物；論述了新興的熒光標(biāo)記物低毒性ZnS:Mn量子點(diǎn)的制備方法和應(yīng)用現(xiàn)狀；簡(jiǎn)述了SEB的危害以及現(xiàn)有的檢測(cè)方法。 2.制備了水溶性CdTe量子點(diǎn)及其磁性熒光納米復(fù)合物；對(duì)相關(guān)性能進(jìn)行了表征；利用磁性熒光復(fù)合物和熒光染料建立了快速檢測(cè)SEB毒素的FRET分析體系。 3.采用水相共沉淀法制備了Mn摻雜的ZnS量子點(diǎn)；X射線衍射和透射電鏡測(cè)試結(jié)果表明該量子點(diǎn)具有閃鋅礦型結(jié)構(gòu),為直徑19nm的球型顆粒；熒光測(cè)試表明所制備的量子點(diǎn)具有窄而對(duì)稱的發(fā)射峰,發(fā)光波長(zhǎng)在580nm左右,為耀眼的橙色光。 4.結(jié)合磁性納米粒子的免疫磁分離和ZnS:Mn量子點(diǎn)的免疫熒光標(biāo)記,采用“夾心式三明治”免疫分析法對(duì)SEB毒素進(jìn)行了檢測(cè),最低檢測(cè)限為0.01ng/mL,具有較好的靈敏度。 5.建立了一種以硝酸纖維素膜為抗體固定載體的熒光免疫分析方法；該方法簡(jiǎn)便快速,具有一定的靈敏度(檢測(cè)限為0.1ng/mL)。
[Abstract]:Staphylococcus aureus enterotoxin BPSEB is a common food toxin produced by Gram-positive cocci. It is a superantigen. SEB plays a very important role in the process of inflammation. It is also an important allergen in allergic diseases, due to its inherent high stability, strong toxicity and easy to spread characteristics. SEB is also a potential biological weapon, so it is very important to develop a rapid, simple and sensitive SEB detection method. Fluorescent immunoassay has high sensitivity and specificity. It is a widely used biological and biochemical analysis technology. Therefore, this paper is based on the previous research in our laboratory. Firstly, a fluorescence resonance energy transfer analysis system using CdTe quantum dots as energy donor and fluorescent dye Alexa Fluor568 as energy receptor was established. But considering the toxicity of CD series quantum dots, the fluorescence immunoassay based on ZnS doped mn low toxicity quantum dots was further established. It is applied to the rapid and sensitive detection of toxin SEB in order to meet the urgent need of food safety monitoring. 1. The application of fluorescent immunoassay (FIA) and the different fluorescent markers commonly used in fluorescence analysis at present are reviewed. The preparation methods and application status of new fluorescent labeled low toxicity ZnS:Mn quantum dots are reviewed in this paper. The harm of SEB and the existing detection methods are briefly described. 2. The water-soluble CdTe quantum dots and their magnetic fluorescent nanocomposites were prepared. The related properties were characterized. A FRET analysis system for rapid detection of SEB toxin was established by using magnetic fluorescent complex and fluorescent dye. 3. Mn-doped ZnS quantum dots were prepared by water-phase coprecipitation. The results of X-ray diffraction and transmission electron microscopy show that the quantum dots have sphalerite structure and are spherical particles with a diameter of 19 nm. Fluorescence measurements show that the quantum dots have a narrow and symmetrical emission peak and the luminescence wavelength is about 580 nm. 4. Combined with the immunomagnetic separation of magnetic nanoparticles and the immunofluorescence labeling of ZnS:Mn quantum dots, the SEB toxin was detected by sandwich immunoassay. The lowest detection limit is 0.01 ng / mL, which has good sensitivity. 5. A fluoroimmunoassay method based on nitrocellulose membrane was established, which is simple, rapid and sensitive (detection limit is 0.1ng / mL ~ (-1)).
【學(xué)位授予單位】：湖南師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：O657.3;R114

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