中文摘要：

目的：研制能够同时检测乙型肝炎表面抗原（HBsAg）野生株和多种变异株的单克隆抗体（mAb）,将筛选出的mAb进行纯度、免疫学性状鉴定,并对其应用效果及质量做初步评价。方法：用中国乙型肝炎病毒感染者血清中分离的HBsAg免疫小鼠,利用杂交瘤细胞融合技术制备抗HBsAg野生株和多种变异株的mAb,将筛选出的mAb经饱和硫酸铵纯化后通过聚丙烯酰胺凝胶电泳、琼脂糖凝胶电泳、ELISA鉴定其纯度、特性,初步评价其应用效果及质量。结果：获得了一株能够同时检测HBsAg野生株和多种变异株的mAb,命名为D12。其纯度较高,特异性强,灵敏度好,Ig亚类测定结果为IgG1,识别位点存在于自然抗原上,其检测HBsAg野生株的能力优于现行3种国产试剂盒,检测HBsAg变异株的能力明显优于现行5种国产试剂盒。结论：成功研制了可以同时识别HBsAg野生株和大多数变异株的mAb,为进一步提高我国当前乙型肝炎病毒变异株的检出率以及加强预防和控制乙型肝炎病毒的传播奠定新的基础。

英文摘要：

Objective： Preparation of monoclonal antibody to detect the wild-type HBsAg as well as variant forms. Methods： BALB/c female Mice were immunized with HBsAg isolated from serum of infected patients. mAbs were prepared by hybridoma technology. Identify the purity and characteristics of the selected mAb purified with saturated ammonium sulfate and give preliminary evaluation of the quality and effectiveness of its application, in which polyacrylamide gel electrophoresis and agarose gel electrophoresis and emzyme-linked immunosorbent assay （ELISA） were used. Results： A monoclonal antibody can detect the wild-type HBsAg as well as variant forms was obtained designated as D12. The purity of D12 was high. D12 was highly sensitive and specific to HBsAg and was tested for subtype as IgGl. The recognition sites existed in the nature antigen. The ability to detect the wild-type HBsAg using D12 was better than that of three commercial HBsAg diagnostic kits and the ability to detect the variant HBsAg using D12 was better than that of five commercial HBsAg diagnostic kits. Conclusion： The preparation of mAb D12 is very important to further improve the detection rate of variant hepatitis B virus and enhance HBV prevention.