抗A型禽流感病毒核蛋白特异性单克隆抗体研究

利用禽流感H9亚型病毒(AIV-H9)免疫Balb/c小鼠,取其脾细胞与骨髓瘤细胞进行融合,经免疫荧光试验(IFA)检测,以研制抗禽流感病毒(AIV)单克隆抗体。结果获得了5株特异性抗AIV核蛋白(NP)的单克隆抗体细胞株,分别命名为AIV-NP-2C3、AIV-NP-6A5、AIV-NP-3H9、AIV-NP-7B4、AIV-NP-2H4。这5株单克隆抗体能与所有试验的AIV-H9病毒反应,Western blotting方法鉴定结果表明,单克隆抗体仅识别60 ku的蛋白抗原,而不与新城疫病毒、禽网状内皮组织增殖症病毒、传染性法氏囊病毒等反应。初步应用结果显示,以这些单克隆抗体建立的间接免疫荧光试验或ELISA方法可迅速检测出禽流感病毒,这些单克隆抗体在禽流感的预防监测中将发挥重要的作用。

Development and characterization of monoclonal antibody specific to nucler protein of avian influenza virus

Five monoclonal antibodies(Mabs) to nucler protein of avian influenza virus (AIV) were developed by fusion between SP2/0 cells and the spleen cells from BALB/c mice immunized with H9 subtype AIV.Specificity of these Mabs were identified by immunofluorescent assay(IFA) and enzyme link immunosorb assay (ELISA).These five Mabs could react with all viruses of AIV-H9 strains in tests,but didn't react with Newcastle disease virus,REV and infectious bursa disease virus.Only a 60 ku protein band of AIV-H9 was recongnized by the Mabs in Western blotting.This suggested that the five Mabs,which were named as AIV-NP-2C3,AIV-NP-6A5,AIV-NP-3H9,AIV-NP-7B4 and AIV-NP-2H4 respectively,were specific against the nucler protein of avian influenza viruses.All avian influenza viruses could be dectected with these Mabs in IFA and ELISA.These results indicate that these Mabs will be potentially useful as immunodiagnostic reagents in future.