奶牛孕酮的无标记阻抗免疫传感器研究
发布时间：2018-08-30 20:38
【摘要】：孕酮(Progesterone)是雌性动物排卵后在黄体和怀孕期胎盘产生的一种天然孕激素。奶牛孕酮的含量随着其繁殖活动而呈现规律性变化,因此,可以通过对孕酮的准确检测诊断奶牛是否发情或妊娠,从而采取受孕措施提高奶牛繁殖效率、增加奶牛业的生产效益。传统的孕酮检测方法有：乳胶凝集抑制试验(LAIT)、酶联免疫吸附测定法(ELISA)、酶免疫测定法(EIA)和放射免疫法(RIA)。但这些方法操作较为复杂,且价格昂贵,不能满足我国畜牧业生产的需求。本课题将孕酮单克隆抗体用蛋白A固定在丝网印刷金电极上,制备了可快速检测孕酮含量的阻抗免疫传感器,并用交流阻抗法进行了孕酮的定量检测,同时设计了一种微弱信号检测装置,取得了较好的效果。本文主要贡献如下：1.为了增加印刷电极的稳定性和灵敏性,对丝网印刷金电极采用电化学方法(在硫酸中进行循环伏安法活化),分析电极活化的最优条件。2.用0.1g/mL的蛋白A修饰丝网印刷金电极并将孕酮单克隆抗体固定在修饰过的电极上制备孕酮阻抗免疫传感器,并进行优化。用制备的孕酮阻抗传感器检测浓度梯度为Ong/mL、1.5625ng/mL、3.125ng/mL、6.25ng/mL、12.5ng/mL、25ng/mL和50ng/mL的孕酮溶液进行交流阻抗法测量,建立阻抗值和孕酮浓度的回归方程,实现对孕酮浓度的定量检测。3.根据试验得到的阻抗数据用Zsimpwin软件拟合出相应的等效电路,并得到电路各元件的值以及拟合误差,进一步对理论模型分析。4.设计了一种微弱信号检测装置,为基于孕酮阻抗免疫传感器制备一种便携式孕酮检测设备打下坚实的基础。
[Abstract]:Progesterone (Progesterone) is a natural progesterone produced after ovulation in the corpus luteum and placenta during pregnancy. The content of progesterone in cows varies regularly with their reproductive activities. Therefore, the accurate detection of progesterone can be used to diagnose whether the cows are estrous or pregnant, so as to improve the reproductive efficiency of cows. Increase the production efficiency of dairy industry. The traditional methods of progesterone detection are: latex agglutination inhibition test (LAIT), enzyme linked immunosorbent assay (LAIT), enzyme immunoassay (EIA) and radioimmunoassay (RIA). However, these methods are complex and expensive, and can not meet the needs of animal husbandry production in China. In this paper, the monoclonal antibody of progesterone was immobilized on the gold electrode of screen printing with protein A, and an impedance immunosensor for rapid detection of progesterone was prepared, and the quantitative detection of progesterone was carried out by AC impedance method. At the same time, a weak signal detection device is designed, and good results are obtained. The main contributions of this paper are as follows: 1. In order to increase the stability and sensitivity of the printing electrode, the electrochemical method (cyclic voltammetry activation in sulfuric acid) was used to analyze the optimum conditions of electrode activation. The 0.1g/mL protein A was used to modify the screen printing gold electrode and the progesterone monoclonal antibody was immobilized on the modified electrode to prepare a progesterone impedance immunosensor. The progesterone solution with the concentration gradient of Ong/mL,1.5625ng/mL,3.125ng/mL,6.25ng/mL,12.5ng/mL,25ng/mL and 50ng/mL was measured by the progesterone impedance sensor. The regression equation of the impedance value and the concentration of progesterone was established, and the quantitative detection of progesterone concentration was realized. According to the impedance data obtained from the experiment, the corresponding equivalent circuit is fitted with Zsimpwin software, and the values of each component of the circuit and the fitting error are obtained, and the theoretical model is further analyzed. A weak signal detection device is designed, which lays a solid foundation for the preparation of a portable progesterone detection device based on progesterone impedance immunosensor.
【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823
本文编号：2214224
[Abstract]:Progesterone (Progesterone) is a natural progesterone produced after ovulation in the corpus luteum and placenta during pregnancy. The content of progesterone in cows varies regularly with their reproductive activities. Therefore, the accurate detection of progesterone can be used to diagnose whether the cows are estrous or pregnant, so as to improve the reproductive efficiency of cows. Increase the production efficiency of dairy industry. The traditional methods of progesterone detection are: latex agglutination inhibition test (LAIT), enzyme linked immunosorbent assay (LAIT), enzyme immunoassay (EIA) and radioimmunoassay (RIA). However, these methods are complex and expensive, and can not meet the needs of animal husbandry production in China. In this paper, the monoclonal antibody of progesterone was immobilized on the gold electrode of screen printing with protein A, and an impedance immunosensor for rapid detection of progesterone was prepared, and the quantitative detection of progesterone was carried out by AC impedance method. At the same time, a weak signal detection device is designed, and good results are obtained. The main contributions of this paper are as follows: 1. In order to increase the stability and sensitivity of the printing electrode, the electrochemical method (cyclic voltammetry activation in sulfuric acid) was used to analyze the optimum conditions of electrode activation. The 0.1g/mL protein A was used to modify the screen printing gold electrode and the progesterone monoclonal antibody was immobilized on the modified electrode to prepare a progesterone impedance immunosensor. The progesterone solution with the concentration gradient of Ong/mL,1.5625ng/mL,3.125ng/mL,6.25ng/mL,12.5ng/mL,25ng/mL and 50ng/mL was measured by the progesterone impedance sensor. The regression equation of the impedance value and the concentration of progesterone was established, and the quantitative detection of progesterone concentration was realized. According to the impedance data obtained from the experiment, the corresponding equivalent circuit is fitted with Zsimpwin software, and the values of each component of the circuit and the fitting error are obtained, and the theoretical model is further analyzed. A weak signal detection device is designed, which lays a solid foundation for the preparation of a portable progesterone detection device based on progesterone impedance immunosensor.
【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823
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