基于极大角度倾斜光纤光栅的猪圆环病毒Ⅱ型检测技术研究
发布时间：2019-07-03 19:25
【摘要】：生化免疫传感器一直是传感器技术领域的一个非常活跃的研究课题,特别是近几年来,随着各种新原理新技术的发展和出现,生化免疫传感器正朝着微型化、集成化、智能化等方向发展。目前,传统的生化免疫检测方法因其检测时间长、测试费用高、仪器体积大、价格昂贵、专业性强等特点,不利于微型化、集成化、实时在线监测的需求。而光纤光栅传感器不但具有体积小、抗干扰、耐腐蚀等优点,而且具有“传”与“感”的同时传输等特点,在疾病诊断、抗体抗原检测和基因测试分析等方面显示出广泛的应用前景。本文以极大角度倾斜光纤光栅Ex-TFG为生化免疫检测平台,用金黄色葡萄球菌A蛋白(SPA)修饰光栅表面,再将猪圆环病毒II型(PCV2)单克隆抗体固定于光栅表面的分子层,用于PCV2抗原的特异性检测。通过光谱数据分析、免疫学原理、光纤表面的表征等方法来研究、分析该生化免疫传感器的性能。本文的主要研究内容包括以下几点:(1)设计并制作Ex-TFG,通过实验检测传感器的折射率灵敏度,建立其谐振波长与折射率的响应关系。(2)研究Ex-TFG的表面修饰方法,并用扫描电镜、荧光显微镜等工具对表面修饰效果进行鉴定。结果表明,传感器表面结合的位点是制约着检测范围和检测极限等参数的关键因素。(3)搭建生化免疫检测系统,对基于Ex-TFG的PCV2生化免疫传感器的最低检测极限、饱和度、检测范围等参数进行研究,结果分别为~10 TCID 50/mL、~4800 TCID 50/mL、10-4800 TCID 50/m L。此外,研究对生化免疫传感器表面的PCV2单克隆抗体的解离方法,获得了良好的重复性。(4)结合实验原理和实验数据,对检测结果进行米氏拟合,建立米氏方程模型,为光纤光栅生化免疫传感器的检测机理提供有力的分析手段。本课题研究的基于Ex-TFG的PCV2新型生化免疫传感器,是生化免疫检测领域的一大补充,具有快速检测、操作简单、微尺寸及痕量检测的优点,为生物学、医学、食品、环境等方面免疫检测提供了一种新的手段。
[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，
本文编号：2509611
[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，
本文编号：2509611
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