非洲猪瘟病毒蛋白p54原核真核表达、间接ELISA检测方法建立及单克隆抗体制备-生物化学与分子生物学专业论文.docxVIP

暨南大学硕士学位论文 暨南大学硕士学位论文 PAGE PAGE VI eukaryotic expression system expressed protein p54 respectly, the ASF standard positive serum can be detected after diluting 1: 1280 and 1:320 dilution by two ELISA respectly, inter and intra CV is less than 10% and 15%. The coincidence rate between commercial ELISA kit and two indirect ELISA is 90.1% and 84.4%. Specificity of the two methods are 100%. The sensitivity of the two method is 88.2% and 80.3% respectly. Conclusion: In this study, two indirect ELISAs detecting ASF antibody in serum were established. The two methods are sensitive, specific, stable and easy to operate. Meanwhile, the p54 monoclonal antibody was generated. The achievement of this study will provide a technical preparation for detection of ASF. Keywords: ASFV, p54, indirect ELISA, monoclonal antibody 英文缩写表 英文缩写 英文全称 中文全称 ASF African swine fever 非洲猪瘟 ASFV African swine fever virus 非洲猪瘟病毒 ELISA enzyme linked immunosorbent assay 酶联免疫吸附检测 OIE Office International Des Epizooties 世界动物卫生组织 BSA bovine serum albumin 牛血清白蛋白 HAD haemadsorption 红细胞吸附 FAT fluorescence antibody test 荧光抗体检测 PCR polymerase chain reaction 聚合酶链式反应 CFS classic swine fever 猪瘟 E.coil Escherichia coli 大肠杆菌 M mol/L 摩尔每升 O.erraticus Ornithodoros erraticus 游走钝缘蜱 O.moubata Ornithodoros moubata 非洲钝缘蜱 PBS phosphate buffer saline 磷酸盐缓冲液 TMB 3,3,5,5-Tetramethylbenzidine 3,3,5,5- HYPERLINK /view/150495.htm 四甲基联苯胺 LAMP loop-mediated isothermal amplification 圆环介导的等温扩增 P positive value 阳性值 N negative value 阴性值 PC positive cut off 阳性阈值 NC negative cut off 阴性阈值 SD standard deviation 标准差 CV coefficient of variation 变异系数 LOD Limit of detection 检测限 目录 摘要 I Abstract II HYPERLINK \l _bookmark0 英文缩写表IV HYPERLINK \l _bookmark1 第一章 前言1 HYPERLINK \l _bookmark2 1.非洲猪瘟的流行趋势1 HYPERLINK \l _bookmark3 2.非洲猪瘟的病理学特征3 HYPERLINK \l _bookmark4 2.1 非洲猪瘟的临床特征3 HYPERLINK \l _bookmark5 2.2 非洲猪瘟常见宿主3 HYPERLINK \l _bookmark6 2.3 非洲猪瘟病的传播途径5 HYPERLINK \l _bookmark7 3. 非洲猪瘟病毒形态学特征和主要组成蛋白6 HYPERLINK \l _bookmark8 4.非洲猪瘟检测方法