【摘要】 目的:获得特异针对禽流感H9亚型血凝素单克隆抗体。方法:以H9N2亚型病毒制成疫苗免疫BALB/c小鼠,取脾细胞与SP2/0细胞融合,取细胞上清用HI筛选。结果:共获得9株稳定分泌针对血凝素蛋白抗体的杂交瘤细胞;经广谱性和特异性鉴定,所有单抗株均能与所有检测的82株不同年代不同地方不同禽源H9亚型流感病毒大部分分离株反应,且所有单抗株均不与非AⅣ-H9病毒反应,而其中一株(8E6)能与所有检测的H9亚型病毒株反应。进行Western blot分析显示, 其中8株单抗能与AⅣ的Mr为75 000处反应,表明是针对AⅣ H9 HA的单抗。结论:本试验获得9株广谱性特异性很好的单抗,特别是8E6株;这些单抗株为以后作病毒抗原性位点分析、临床检测和流行病学调查提供了良好试剂。更多还原

【Abstract】 Objective:To development monoclonal antibody ( McAb) specific to hemagglutinin of H9 subtype avian influenza viruses( AⅣ). Methods: Fusing SP2/ O-Ag-14 myeloma cells with spleen cells of BALB/ c mice immunized with live AⅣ-H9 subtype vaccine, and positive hybridoma clones were screened by HI test with supernatant. The specificity of McAbs was characterised by HI test ,ELISA and Western blot. Results:Nine stains McAb against HA of AIV H9 subtype viruses were obtained. All of these only specifically reaction with most of 82 strains of subtype AⅣ-H9 viruses, which isolated in chicken and duck in different provinces in different year, but not with other viruses. Specificity analysis by Western blot confirm that eight of these McAbs recognize HA, and one of the nine McAbs reaction with all of the detected H9 subtype viruses. Conclusion: Nine McAbs specific to hemagglutinin of AⅣ-H9 were obtained, which provided useful tool for virus antigenic epitope analysis .clinical diagnosis and epidemiological investigation.更多还原