用基因疫苗制备H9亚型禽流感病毒单克隆抗体  

Preparation of Monoclonal Antibodies against the Hemagglutinin of H9 Subtype of Avian Influenza Virus by Genetic Immunization

作　　者：詹爱军[1,2] 王新卫[2,3] 王敬师[2] 李少璃[2] 于康震[4] 毕英佐[2] 

机构地区：[1]扬州大学,扬州225009 [2]华南农业大学动物科学学院,广州510642 [3]河南农业大学牧医工程学院,郑州450002 [4]中国兽药监察所,北京100081

出　　处：《生物技术通报》2009年第1期117-120,共4页Biotechnology Bulletin

基　　金：广东省科技重大专项计划项目(2004A2090102)

摘　　要：用H9亚型禽流感病毒(AIV)HA基因反转录cDNA第一链PCR扩增其HA基因,PCR产物与pcDNA3.1(+)质粒构建重组质粒作为基因疫苗免疫8周龄Balb/C小鼠,细胞融合后用血凝抑制试验(HI)和ELISA试验检测细胞培养上清,各获得一株阳性细胞株,经3次亚克隆后都能稳定分泌H9AIV特异性抗体。特异性检测与NDV、H5亚型AIV、产蛋下降综合症(EDS76)病毒毒株没有反应,2株单抗经亚型鉴定均为IgG2b,轻链的亚型为kappa链。所获得的单克隆抗体将在禽流感快速诊断和疾病预警监控中发挥重要作用。Hemagglutinin genes of H9 subtype of avian influenza virus （AIV） was amplified by PCR.The digested PCR products of HA was cloned into plasmid pcDNA3.1 （＋）. BALB/C mice of 8 weeks old were immunized with recombinant plasmid pcDNA-H9. The spleen cells of immunized mice were fused with Sp2/0 myeloma cells. Indirect enzyme linked immunosorbent assay（ELISA） and hemagglutinatin inhibition（HI） test were used to screen hybridoma cells. Two McAbs specifically against AIV H9 were obtained,which were not reacted to AIV-H5,NDV and EDS76 virus. The subtype of the belonging to McAbs was identified as IgG2b,kappa light chain. The two McAbs would be useful as specific reagents in diagnosing avian influenza in practical poultry industry.

关 键 词：H9亚型禽流感病毒 血凝素 单克隆抗体 基因免疫 

分 类 号：S852.5[农业科学—基础兽医学]