新城疫病毒单克隆抗体的制备及其与不同分离株的反应性  

Preparation of Monoclonal Antibodies Against Newcastle Disease Virus and Their Reactivity Against Different Isolates

作　　者：徐璇[1] 叶云[1] 梁明征[1] 吴唯维[1] 刘珊[1] 吴华静 何秀苗[1,2] 韦天超[3] 韦平[3] 

机构地区：[1]广西民族大学海洋与生物技术学院,广西南宁530006 [2]广西林产化学与工程重点实验室,广西南宁530006 [3]广西大学,广西南宁530005

出　　处：《中国家禽》2013年第24期24-27,共4页China Poultry

基　　金：广西自然科学基金项目(2012GXNSFAA053060);广西科技攻关项目(桂科攻1123007-4);广西高校科学技术研究项目(2013YB069);南宁市科技攻关项目(201001021B;200901028B)

摘　　要：以纯化后的新城疫病毒(Newcastle disease virus,NDV)强毒参考株F48E8为免疫原,腹腔注射法免疫BALB/c小鼠。取免疫后小鼠血清中血凝抑制(HI)抗体滴度最高的脾脏细胞与骨髓瘤细胞SP2/0进行融合,应用HI试验检测融合细胞的上清筛选阳性克隆。结果显示,获得了4株能稳定传代并分泌抗NDV血凝素(HN)单克隆抗体的杂交瘤细胞,分别命名为1G10、2H5、5B9和5E10。小鼠接种105个左右杂交瘤细胞时获得的腹水效价达到29.5～211,特异性试验表明,所制备单克隆抗体仅与NDV F48E8、La Sota和C30株发生特异性反应,而与其他常见禽类病毒不出现阳性反应。此外,所制备的单克隆抗体5E10可通过间接免疫荧光法检测Vero细胞上的NDV感染,并可通过HI试验用于区分鸡源和鸽源NDV分离株。BALB/c mice were immunized with purified Newcastle disease virus （NDV）reference strain F48E8 via abdominal cavity injection. Splenocytes from the immunized BALB/c mice were fused with SP2/O myeloma cells. The positive hybridoma clones were screened by hemagglutination inhibition （HI）test with the supernatant,and four monoclonal antibodies （Mabs） against HN protein of NDV was obtained, namely 1GIO, 2H5,5B9 and 5El0, respectively. The results showed that HI titer of the ascites could reach to 29.5 to 2 when the mice were injected with about 105 hybridoma ceils. Specificity tests indicated that the monoclonal antibodies were specific to F48E8,La Sota and C30 strain, but not reacted with any of the other avian viruses. Furthermore,5ElO could be used to detect NDV in Vero cells by in- direct fluorescence assay, and differentiate chicken strain from pigeon strains in NDV.

关 键 词：新城疫病毒 血凝素 单克隆抗体 间接免疫荧光法 

分 类 号：S852.43[农业科学—基础兽医学]