禽网状内皮组织增生症病毒单抗制备及在外源病毒检测中的应用  

Preparation of Monoclonal Antibody against Avian Reticuloendotheliosis Virus and Its Application in Exogenous Viruses Detection

作　　者：陈晓春[1] 赵炜 苏佳 王嘉[1] 侯力丹[1] 黄小洁[1] 吴华伟[1] 李俊平[1] CHEN Xiao-chun;ZHAO Wei;SU Jia;WANG Jia;HOU Li-dan;HUANG Xiao-jie;WU Hua-wei;LI Jun-ping(China Institute of Veterinary Drug Control, Beijing 100081,China)

机构地区：[1]中国兽医药品监察所,北京100081

出　　处：《中国兽药杂志》2022年第6期1-8,共8页Chinese Journal of Veterinary Drug

基　　金：中国兽医药品监察所“兽药行业公益性重点专项”(GY202012)。

摘　　要：将禽网状内皮组织增生症病毒(REV)囊膜蛋白gp90(SU)的基因合成后连接至原核表达载体中进行表达,获得REV-SU蛋白,通过Ni柱亲和层析获得纯化的重组蛋白,免疫BALB/c小鼠,应用杂交瘤技术获得1株分泌特异性抗REV-SU蛋白的单克隆抗体杂交瘤细胞系(2A2株),该细胞株制备的腹水与不同REV毒株具有良好的反应性,荧光效价可达1∶51200,与不同禽白血病病毒毒株和其他常见禽的病原均没有交叉反应,特异性良好。应用该单克隆抗体建立的间接免疫荧光法能检出至少5 TCID_(50)REV感染,与多克隆抗体作为检测一抗具有同等的效力,且检测背景更加纯净,适用于外源性REV检验。To establish an indirect immunofluorescence staining method for detecting exogenous viruses,the gene of the envelope protein gp90(SU)of avian reticuloendotheliosis virus(REV)was synthesized and ligated into prokaryotic expression vector for expression,and REV-SU protein was obtained in this study.Then,the purified recombinant protein was obtained by Ni affinity chromatography.After immunizing BALB/c mice,a hybridoma cell line(strain 2A2)secreting specific monoclonal antibody against REV-SU protein was obtained with hybridoma technique.The ascites prepared using this cell line had high reactivity with different REV strains,with the immunofluorescence titer reaching 1∶51200,but no cross-reaction with different avian leukosis virus strains or other conventional avian viruses,showing high specificity.The indirect immunofluorescence assay established with the monoclonal antibody secreted by this cell line could detect more than 5 TCID_(50) REV,as the same effect as the detection based on polyclonal antibody,and the detection background was more pure.The results confirm that this method is suitable for the detection of exogenous REV.

关 键 词：禽网状内皮组织增生症病毒 外源病毒检验 gp90 单克隆抗体 间接免疫荧光法 

分 类 号：S852.65[农业科学—基础兽医学]