本文選題：內(nèi)皮抑素 + 膠乳增強免疫比濁法�。� 參考：《南京大學》2015年碩士論文

【摘要】：目前國內(nèi)的內(nèi)皮抑素濃度的檢測方法主要是酶聯(lián)免疫吸附試驗法,但是該方法耗時且價格昂貴。為快速簡便的檢測內(nèi)皮抑素,考慮到膠乳增強免疫比濁法具有特異性好和靈敏度高的特點,同時能減少非特異性凝集,假陽性率較低,并在醫(yī)院的臨床檢驗中能廣泛應用等優(yōu)點。因此本課題根據(jù)膠乳增強比濁法的基本原理,旨在研究開發(fā)出質(zhì)優(yōu)價低的國產(chǎn)重組人內(nèi)皮抑素檢測試劑盒,這對于減輕患者負擔具有重要意義。研究采用方法是將實驗室原采用的重組人內(nèi)皮抑素兔抗體(IgG)改為雞抗體(IGY)標記合適粒徑的膠乳,以合適的吸光度范圍和較大的線性范圍為標準。優(yōu)化選擇PEG6000的最適濃度、檢測波長、加樣量等條件,并繪制標準曲線,確定檢測的線性范圍。實驗結(jié)果表明,初步制備了膠乳增強免疫比濁法內(nèi)皮抑素檢測試劑盒,為重組人內(nèi)皮抑素的科研和臨床檢測提供了新的選擇。糖尿病(DM)是一種常見的內(nèi)分泌代謝性疾病,DM患者由于長期不良的血糖控制從而導致慢性并發(fā)癥,因此能夠嚴格地控制血糖水平是治療糖尿病和預防各種并發(fā)癥的有效手段。目前可用作血糖控制的指標主要是糖化白蛋白(GA)和糖化血紅蛋白(HbAlc)。研究發(fā)現(xiàn)糖化白蛋白(GA)檢測較適合妊娠期糖尿病的診斷和療效評估。最早測定GA的方法是高壓液相離子交換法,該方法具有準確性高的優(yōu)點,但是代價高,處理樣本量小,不適宜臨床常規(guī)開展,故未能得到廣泛應用。后來日本旭化成制藥株式會社研制開發(fā)了液態(tài)試劑,酮胺氧化酶法(KAOD法)檢測GA,該方法簡單易操作,重復性較高,還可以適用于全自動生化分析儀。我們從方法學上改進仿制了糖化白蛋白酶法檢測試劑盒并進行初步的研究。
[Abstract]:Enzyme-linked immunosorbent assay (Elisa) is the main method for the detection of endostatin concentration in China, but it is time-consuming and expensive. In order to detect endostatin quickly and easily, considering that latex enhanced immunoturbidimetry has the characteristics of good specificity and high sensitivity, it can reduce nonspecific agglutination, and the false positive rate is low. And can be widely used in the hospital clinical examination and other advantages. Therefore, according to the basic principle of latex enhanced turbidimetry, the purpose of this paper is to develop a domestic recombinant human endostatin kit with low quality and low value, which is of great significance to lighten the burden of patients. The method used in this study was to change the recombinant human endostatin rabbit antibody IgG into chicken antibody IGY to label the latex with appropriate particle size, with the standard of suitable absorbance range and large linear range. The optimal concentration, detection wavelength and sample amount of PEG6000 were optimized, and the standard curve was drawn to determine the linear range of detection. The results showed that a latex enhanced immunoturbidimetric kit for endostatin detection was prepared, which provides a new choice for the scientific research and clinical detection of recombinant human endostatin. Diabetic diabetes mellitus (DMN) is a common endocrine and metabolic disease patients with diabetes caused by chronic complications due to long-term poor control of blood sugar, so it is an effective method to treat diabetes mellitus and prevent various complications by strictly controlling blood glucose level. At present, glycosylated albumin (GAA) and glycosylated hemoglobin (HbAlc) can be used to control blood sugar. Glycosylated albumin (GAA) was found to be more suitable for the diagnosis and evaluation of gestational diabetes mellitus (GDM). The earliest method for the determination of GA is high pressure liquid phase ion exchange method, which has the advantages of high accuracy, but high cost, small sample size and unsuitable for clinical routine development, so it has not been widely used. The liquid reagent, keto-amine oxidase method and KAOD method were developed to detect GA. this method is simple and easy to operate, has high reproducibility, and can also be used for automatic biochemical analyzer. We improved and studied the saccharified white protease assay kit in methodology.
【學位授予單位】：南京大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R446.6

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