内皮抑素胶乳增强免疫比浊法检测试剂盒和糖化白蛋白酶法检测试剂盒的研究

发布时间：2018-05-24 01:22

本文选题：内皮抑素 + 胶乳增强免疫比浊法　；参考：《南京大学》2015年硕士论文

【摘要】：目前国内的内皮抑素浓度的检测方法主要是酶联免疫吸附试验法,但是该方法耗时且价格昂贵。为快速简便的检测内皮抑素,考虑到胶乳增强免疫比浊法具有特异性好和灵敏度高的特点,同时能减少非特异性凝集,假阳性率较低,并在医院的临床检验中能广泛应用等优点。因此本课题根据胶乳增强比浊法的基本原理,旨在研究开发出质优价低的国产重组人内皮抑素检测试剂盒,这对于减轻患者负担具有重要意义。研究采用方法是将实验室原采用的重组人内皮抑素兔抗体(IgG)改为鸡抗体(IGY)标记合适粒径的胶乳,以合适的吸光度范围和较大的线性范围为标准。优化选择PEG6000的最适浓度、检测波长、加样量等条件,并绘制标准曲线,确定检测的线性范围。实验结果表明,初步制备了胶乳增强免疫比浊法内皮抑素检测试剂盒,为重组人内皮抑素的科研和临床检测提供了新的选择。糖尿病(DM)是一种常见的内分泌代谢性疾病,DM患者由于长期不良的血糖控制从而导致慢性并发症,因此能够严格地控制血糖水平是治疗糖尿病和预防各种并发症的有效手段。目前可用作血糖控制的指标主要是糖化白蛋白(GA)和糖化血红蛋白(HbAlc)。研究发现糖化白蛋白(GA)检测较适合妊娠期糖尿病的诊断和疗效评估。最早测定GA的方法是高压液相离子交换法,该方法具有准确性高的优点,但是代价高,处理样本量小,不适宜临床常规开展,故未能得到广泛应用。后来日本旭化成制药株式会社研制开发了液态试剂,酮胺氧化酶法(KAOD法)检测GA,该方法简单易操作,重复性较高,还可以适用于全自动生化分析仪。我们从方法学上改进仿制了糖化白蛋白酶法检测试剂盒并进行初步的研究。
[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.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.6

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