H9N2亚型禽流感病毒血凝素单克隆抗体的制备及生物学特性分析  

Generation and Characterization of Monoclonal Antibodies Against Hemagglutinin of H9N2 Subtype Avian Influenza Virus

作　　者：李佳昕 孙文强[1] 袁水庆 王晓泉[1,2] 刘晓文[1,2] 刘秀梵[1,2] 

机构地区：[1]扬州大学兽医学院动物传染病实验室,江苏扬州225009 [2]江苏省人兽共患病学重点实验室,江苏扬州225009

出　　处：《中国家禽》2017年第21期19-23,共5页China Poultry

基　　金："十二五"国家科技支撑计划项目(2015BAD12B01-3)

摘　　要：1994年以来,H9N2亚型禽流感病毒(AIV)在中国广泛流行,给养禽业造成了巨大损失。病毒已表现出明显的遗传和抗原多样性,但是对于其抗原性的演化研究较少。本研究选取2014年H9N2亚型AIV分离株A/chicken/Jiangsu/TM118/2014制备针对其血凝素的单克隆抗体,共获得8株稳定分泌针对血凝素蛋白的杂交瘤细胞,分别命名为4B6、3F2、6D11、6B5、1H6、1G6、2D11、3D2,腹水HI效价在2^(11)~2^(19)之间;特异性鉴定结果显示,8株单克隆抗体仅特异针对H9亚型禽流感病毒,而与其他亚型禽流感病毒及其他感染家禽的常见病毒不反应;广谱性分析显示,6D11可以与所选毒株的绝大部分反应,其它7株与不同年代、不同谱系的反应性不同;8株单克隆抗体均能与感染H9亚型禽流感病毒的MDCK细胞特异性反应,呈现绿色荧光。本研究所获得的单克隆抗体可在H9亚型禽流感病毒的抗原性位点分析、流行病学监测及临床检测中发挥重要作用。H9N2 subtype avian influenza virus （AIV）has been circulating in China since 1994, and caused significant losses in chicken flocks in recent years. H9N2 viruses display a remarkable genetic and antigenic diversity. However, there is a limited understanding of antigenic characteristics of the viruses. We prepared eight monoclonal antibodies （McAbs） against the heamagglutinin of the virus using H9N2 strain A/ehicken/Jiangsu/TMll8/2014, which were named as 4B6, 3F2, 6D11, 6B5, 1H6, 1G6, 2D11, 3D2. The HI titers in ascites were between 2H to 219. Specificity test showed that eight McAbs reacted only with H9 subtype AIV, but not with other subtypes of AIV and other avian viruses. The broad spectrum analysis showed that 6Dll could react with most of the selected H9N2 AIV. The other 7 strains had various reactivity profile with strains from different years. All McAbs were able to react specifically with H9N2 virus-infected MDCK cells. The monoelonal antibody obtained in this study have demonstrated utility as antigenic analysis, epidemiological monitoring and clinical detection.

关 键 词：H9N2 禽流感 单克隆抗体 

分 类 号：S855.3[农业科学—临床兽医学]