【摘要】：生化免疫傳感器一直是傳感器技術(shù)領(lǐng)域的一個(gè)非常活躍的研究課題,特別是近幾年來,隨著各種新原理新技術(shù)的發(fā)展和出現(xiàn),生化免疫傳感器正朝著微型化、集成化、智能化等方向發(fā)展。目前,傳統(tǒng)的生化免疫檢測(cè)方法因其檢測(cè)時(shí)間長(zhǎng)、測(cè)試費(fèi)用高、儀器體積大、價(jià)格昂貴、專業(yè)性強(qiáng)等特點(diǎn),不利于微型化、集成化、實(shí)時(shí)在線監(jiān)測(cè)的需求。而光纖光柵傳感器不但具有體積小、抗干擾、耐腐蝕等優(yōu)點(diǎn),而且具有“傳”與“感”的同時(shí)傳輸?shù)忍攸c(diǎn),在疾病診斷、抗體抗原檢測(cè)和基因測(cè)試分析等方面顯示出廣泛的應(yīng)用前景。本文以極大角度傾斜光纖光柵Ex-TFG為生化免疫檢測(cè)平臺(tái),用金黃色葡萄球菌A蛋白(SPA)修飾光柵表面,再將豬圓環(huán)病毒II型(PCV2)單克隆抗體固定于光柵表面的分子層,用于PCV2抗原的特異性檢測(cè)。通過光譜數(shù)據(jù)分析、免疫學(xué)原理、光纖表面的表征等方法來研究、分析該生化免疫傳感器的性能。本文的主要研究?jī)?nèi)容包括以下幾點(diǎn):(1)設(shè)計(jì)并制作Ex-TFG,通過實(shí)驗(yàn)檢測(cè)傳感器的折射率靈敏度,建立其諧振波長(zhǎng)與折射率的響應(yīng)關(guān)系。(2)研究Ex-TFG的表面修飾方法,并用掃描電鏡、熒光顯微鏡等工具對(duì)表面修飾效果進(jìn)行鑒定。結(jié)果表明,傳感器表面結(jié)合的位點(diǎn)是制約著檢測(cè)范圍和檢測(cè)極限等參數(shù)的關(guān)鍵因素。(3)搭建生化免疫檢測(cè)系統(tǒng),對(duì)基于Ex-TFG的PCV2生化免疫傳感器的最低檢測(cè)極限、飽和度、檢測(cè)范圍等參數(shù)進(jìn)行研究,結(jié)果分別為~10 TCID 50/mL、~4800 TCID 50/mL、10-4800 TCID 50/m L。此外,研究對(duì)生化免疫傳感器表面的PCV2單克隆抗體的解離方法,獲得了良好的重復(fù)性。(4)結(jié)合實(shí)驗(yàn)原理和實(shí)驗(yàn)數(shù)據(jù),對(duì)檢測(cè)結(jié)果進(jìn)行米氏擬合,建立米氏方程模型,為光纖光柵生化免疫傳感器的檢測(cè)機(jī)理提供有力的分析手段。本課題研究的基于Ex-TFG的PCV2新型生化免疫傳感器,是生化免疫檢測(cè)領(lǐng)域的一大補(bǔ)充,具有快速檢測(cè)、操作簡(jiǎn)單、微尺寸及痕量檢測(cè)的優(yōu)點(diǎn),為生物學(xué)、醫(yī)學(xué)、食品、環(huán)境等方面免疫檢測(cè)提供了一種新的手段。
[Abstract]:Biochemical immunosensors have always been a very active research topic in the field of sensor technology, especially in recent years, with the development and emergence of various new principles and new technologies, biochemical immunosensors are developing towards miniaturization, integration, intelligence and so on. At present, the traditional biochemical immune detection method is not conducive to miniaturization, integration and real-time online monitoring because of its long detection time, high test cost, large instrument volume, high price, strong professionalism and so on. Fiber grating sensor not only has the advantages of small volume, anti-interference, corrosion resistance, but also has the characteristics of "transmission" and "sense" at the same time. It has shown a wide range of application prospects in disease diagnosis, antibody antigen detection and gene testing and analysis. In this paper, the surface of Ex-TFG was modified by Staphylococcus aureus A protein (SPA), and then the monoclonal antibody of swine circovirus type II (PCV2) was fixed on the molecular layer of the grating surface, which was used for the specific detection of PCV2 antigen. The performance of the biochemical immunosensor was analyzed by means of spectral data analysis, immunological principle and optical fiber surface characterization. The main research contents of this paper include the following points: (1) Ex-TFG, is designed and fabricated to detect the refractive index sensitivity of the sensor through experiments, and the response relationship between resonance wavelength and refractive index is established. (2) the surface modification method of Ex-TFG is studied, and the surface modification effect is identified by scanning electron microscope, fluorescence microscope and other tools. The results show that the binding sites on the surface of the sensor are the key factors that restrict the detection range and detection limit. (3) the minimum detection limit, saturation and detection range of PCV2 biochemical immunosensors based on Ex-TFG are studied by building a biochemical immune detection system. The results are as follows: ~ 10 TCID 50 mL, 鈮，

本文編號(hào)：2509611