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目的方法利用ZhouJian教授提供的重组质粒DNApBV220/eAg,转染DH5α细胞,经30℃培养3小时,42℃培养6小时温度诱导后,提取乙型肝炎病毒e抗原(HBeAg)。经DEAE-SepharoseFastFlow层折纯化后免疫打点分析,收集阳性蛋白。经SDS-PAGE电泳,考马斯亮蓝G250染色,显示为单一的蛋白带,用免疫印迹法(Westernblot)和免疫打点(lmmunodot)法,确定表达产物的特异性。将纯化的HGeAg用酶联免疫吸附试验(ELISA)和免疫条。方法基检测人血清中的相关抗体。结果经49例乙型肝炎病毒e抗体(HBeAb)阳性病人血清检验证实了其特异性。结论免疫条方法较ELISA方法更敏感、特异。
Objectives The method uses the recombinant plasmid DNApBV220 / eAg provided by Professor ZhouJian to transfect DH5α cells, which is cultured for 3 hours at 30 ° C. and is induced at a temperature of 42 ° C. for 6 hours to extract the hepatitis B virus e antigen (HBeAg). After DEAE-SepharoseFastFlow layer purification purification immune RBI analysis, collecting positive protein. After SDS-PAGE electrophoresis, Coomassie brilliant blue G250 staining showed a single protein band. The specificity of the expressed product was determined by Western blot and lmmunodot method. Purified HGeAg was subjected to enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Method based detection of human serum related antibodies. The results of 49 cases of hepatitis B virus e antibody (HBeAb) -positive patients serum confirmed its specificity. Conclusion Immuno-strip method is more sensitive and specific than ELISA method.