【摘要】：孕酮(Progesterone)是雌性動物排卵后在黃體和懷孕期胎盤產(chǎn)生的一種天然孕激素。奶牛孕酮的含量隨著其繁殖活動而呈現(xiàn)規(guī)律性變化,因此,可以通過對孕酮的準(zhǔn)確檢測診斷奶牛是否發(fā)情或妊娠,從而采取受孕措施提高奶牛繁殖效率、增加奶牛業(yè)的生產(chǎn)效益。傳統(tǒng)的孕酮檢測方法有：乳膠凝集抑制試驗(yàn)(LAIT)、酶聯(lián)免疫吸附測定法(ELISA)、酶免疫測定法(EIA)和放射免疫法(RIA)。但這些方法操作較為復(fù)雜,且價(jià)格昂貴,不能滿足我國畜牧業(yè)生產(chǎn)的需求。本課題將孕酮單克隆抗體用蛋白A固定在絲網(wǎng)印刷金電極上,制備了可快速檢測孕酮含量的阻抗免疫傳感器,并用交流阻抗法進(jìn)行了孕酮的定量檢測,同時(shí)設(shè)計(jì)了一種微弱信號檢測裝置,取得了較好的效果。本文主要貢獻(xiàn)如下：1.為了增加印刷電極的穩(wěn)定性和靈敏性,對絲網(wǎng)印刷金電極采用電化學(xué)方法(在硫酸中進(jìn)行循環(huán)伏安法活化),分析電極活化的最優(yōu)條件。2.用0.1g/mL的蛋白A修飾絲網(wǎng)印刷金電極并將孕酮單克隆抗體固定在修飾過的電極上制備孕酮阻抗免疫傳感器,并進(jìn)行優(yōu)化。用制備的孕酮阻抗傳感器檢測濃度梯度為Ong/mL、1.5625ng/mL、3.125ng/mL、6.25ng/mL、12.5ng/mL、25ng/mL和50ng/mL的孕酮溶液進(jìn)行交流阻抗法測量,建立阻抗值和孕酮濃度的回歸方程,實(shí)現(xiàn)對孕酮濃度的定量檢測。3.根據(jù)試驗(yàn)得到的阻抗數(shù)據(jù)用Zsimpwin軟件擬合出相應(yīng)的等效電路,并得到電路各元件的值以及擬合誤差,進(jìn)一步對理論模型分析。4.設(shè)計(jì)了一種微弱信號檢測裝置,為基于孕酮阻抗免疫傳感器制備一種便攜式孕酮檢測設(shè)備打下堅(jiān)實(shí)的基礎(chǔ)。
[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.
【學(xué)位授予單位】：山西農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S823

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