摘要
以H5N1亚型虎源流感病毒免疫Balb/c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2/0融合,用间接ELISA试验检测细胞培养上清,获得2株阳性杂交瘤细胞克隆株,命名为3A13和1B8。分别制备腹水后进行纯化,获得了抗H5亚型虎源流感病毒血凝素的单克隆抗体,用制备的单克隆抗体结合胶体金免疫层析技术,制备了H5亚型虎源流感病毒免疫胶体金快速诊断试纸条。该试纸条对H5N1虎源流感病毒鸡胚培养毒、H5亚型禽流感病毒标准抗原,以及24份虎疑似感染H5N1亚型虎源流感病毒感染病料和小鼠模型病料等检测结果为阳性,同时对H7和H9亚型禽流感病毒标准抗原及传染性支气管炎、新城疫抗原等检测结果为阴性；与HA-HI和RT-PCR的平行对比试验表明,试纸条检测敏感度与血凝试验和血凝抑制试验的敏感性相符。本试验所研制的免疫胶体金诊断试纸条可用于H5亚型虎源流感病毒的特异诊断和流行病学调查,为开发成品化检测试纸条奠定基础。
H5N1 tiger-born Influenza A virus was chosen to immunize Balb/c mice in order to produce the Monoclonal antibodies（Mabs） named 3A13 and 1B8. Two McAbs specifically against avian influenza virus （AIV） were obtained. Colloidal gold particles were prepared and labeled to an anti-H5 subtype tiger-born Influenza virus hemagglutinin monoclonal antibody to establish Colloidal-gold test strips for the rapide detection of H5 subtype Influenza virus. AIV H5 antigen, H5 subtype tiger-born Influenza A virus and other subtypes of AIV were detected. The results indicated that this method had high specificity. No cross-reaction was observed in the samples of NDV,IBV and IBDV. The sensitivity of the assay was as same as hemagglutination and hemagglutination inhibition assay. So we think this method may be a good reference in detection of H5 subtype tiger-born Influenza virus when H5 subtype influenza A viruses outbreak.
出处
《动物医学进展》
CSCD
2007年第1期22-25,共4页
Progress In Veterinary Medicine
关键词
单克隆抗体
H5N1亚型虎源流感病毒
免疫胶体金
monoclonal antibodies
H5N1 subtype tiger-born Influenza virus
immune colloidal gold
