犬瘟热病毒单克隆抗体的研制及胶体金试纸条检测方法的建立  

Preparation of Monoclonal Antibody against Canine Distemper Virus and Development of Colloidal Gold Strip

作　　者：张敏 李建 舒金秀[2] 冯超林[2] 石宝兰 郑良益 牟林琳 柏娇 胡玉立 徐松 刘国兴 闵娟娟 石瑛 顾素云 谢红玲 ZHANG Min;LI Jian;SHU Jin-xiu;FENG Chao-lin;SHI Bao-lan;ZHENG Liang-yi;MU Lin-lin;BAI Jiao;HU Yu-li;XU Song;LIU Guo-xing;MIN Juan-juan;SHI Ying;GU Su-yun;XIE Hong-ling(Sinopharm Animal Health Corporation Ltd.,Wuhan 430075,China;Hubei Province Institute of Veterinary Drug,WuHan 430070,China)

机构地区：[1]国药集团动物保健股份有限公司,武汉430075 [2]湖北省兽药监察所,武汉430070

出　　处：《中国兽药杂志》2022年第12期16-24,共9页Chinese Journal of Veterinary Drug

摘　　要：利用原核表达、纯化的犬瘟热病毒(Canine distemper virus, CDV)N蛋白免疫BALB/c小鼠,成功筛选到两株分泌CDV N蛋白单克隆抗体的杂交瘤细胞株,分别命名为D3、D6。2株杂交瘤细胞分泌单克隆抗体敏感性、特异性良好,能识别不同来源的CDV株,单抗亚类均为IgG1。以D3作为金标抗体,D6作为检测抗体,兔抗鼠IgG作为质控线抗体,制备CDV胶体金检测试纸条,检测CDV敏感性为1000 TCID50,能在CDV、犬细小病毒、犬腺病毒2型、狂犬病病毒中有效检测区分出CDV。本研究建立的胶体金试纸条检测方法灵敏度高、特异性强,适用于CDV临床快速诊断。In this study, the prokaryotic expression of canine distemper virus(CDV) N protein was induced, and the purified N protein was immunized to BALB/C mice. Two monoclonal hybridioma cell lines(D3, D6) secreting CDV N protein antibody were developed by hybridioma cell technology. The monoclonal antibodies secreted by the two hybridoma cells were sensitive and specific, and could recognize different canine distemper virus strains. The monoclonal antibody subclass both was IgG1. Using D3 as the colloidal gold labeled antibody, D6 as the detection antibody and rabbit anti-mouse as the quality control antibody, a colloidal gold strip for detecting porcine coronavirus was prepared. The test line was visible with a detection limit of 1000 TCID50(CDV). And the specificity tests of dipsticks showed that could distinguish CDV, canine parvovirus(CPV), canine adenovirus type 2(CAV2) and rabies virus(RV). The colloidal gold strip detection method established in this study had good sensitivity and specificity, and could be suitable for rapid clinical diagnosis of CDV.

关 键 词：犬瘟热病毒 N蛋白 单克隆抗体 胶体金试纸条 

分 类 号：S852.65[农业科学—基础兽医学]