抗-HBs的纯化及在乙肝病毒表面抗原检测中的应用论文材料.docxVIP

PAGE  PAGE 32 抗-HBs的纯化及在乙肝病毒表面抗原检测中的应用 摘要: 本文用硫酸铵饱和沉淀法纯化抗-HBs，建立起一种系统的检测乙肝病毒表面抗原的双抗体夹心ELISA法，从而使得乙肝病毒表面抗原的检测快速简便准确。我们通过对不同浓度抗体的包被，用已知表面抗原阳性血清进行检测，确定抗原抗体反应的最佳包被抗体稀释度是1:500；然后以此稀释度包被抗体，检测80份待检血清，结果发现有4份为表面抗原阳性，余者呈表面抗原阴性，与标准对照板的检测结果一致。另外还对双抗体夹心酶联免疫吸附法检测过的80份待测血清又进行了尝试性的混合样本分析法检测。本研究建立的抗-HBs纯化方案及HBsAg ELISA检测体系在临床医学免疫诊断中具有一定可操作性和实际应用价值。 关键词: 乙型肝炎表面抗原 硫酸铵饱和沉淀法 抗-HBs 酶联免疫吸附试验 双抗体夹心法　 混合样本分析法 Study on purification of anti-HBs and its application in detection of HBsAg Abstract: The anti-HBs were purified by saturated ammonium sulfate precipitation in this paper, and employed to establish double-antibody-sandwich enzyme linked immunosorbent assay (ELISA) for detection of hepatitis B surface antigen. Therefore, we can get a simple，rapid，sensitive and specific ELISA system of examining hepatitis B surface antigen. Different concentrations of antibodies were coated on polystyrene (PS) 96-well plates, and under detection with known HBsAg positive serum in order to determine the optimal antibody dilution which was 1:500, then with dilution antibodies packaged on polystyrene miroplates, 80 serum species were examined, and four HBsAg positive serum specimens were found, the rest were negative for surface antigen. For comparison, four HBsAg positive sera were tested simultaneously in our control experiments. 80 serum samples were also measured by a mixed samples analysis. Anti-HBs purification and the established ELISA system for HBsAg detection in the study maybe have its way in a certain immune workable and practical application value of clinical diagnosis. Key words: hepatitis B surface antigen, saturated ammonium sulfate precipitation, ELISA, anti-HBs, double-antibody-sandwich assay, mixed samples analysis 第一章 前言 1.1 乙型肝炎病毒的生物学特征 1963年外国学者在血友病患者血清中发现有一种抗体，与澳大利亚土著人血清中的一种抗原起交叉反应，这种抗原当时被称为“澳大利亚抗原”[1, 2]。1968年，这种抗原被证实就是乙肝病毒的表面抗原。1970年，Dane用电子显微镜在乙肝患者血清中观察到了完整的乙肝病毒颗粒(即Dane颗粒)以及乙肝病毒表面抗原颗粒。 乙肝病毒属于嗜肝DNA病毒科中哺乳动物病毒。完整的HBV颗粒直径为42 nm，分为包直径22 nm，其本身无传染性，核心部分含有球状双股