| 抗H3N2亚型猪流感病毒单克隆抗体的制备与鉴定 |
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| 引用本文： | 孔维,乔传玲,陈艳,杨焕良,展小过,辛晓光,陈化兰.抗H3N2亚型猪流感病毒单克隆抗体的制备与鉴定[J].畜牧与兽医,2010,42(9). |
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| 作者姓名： | 孔维 乔传玲 陈艳 杨焕良 展小过 辛晓光 陈化兰 |
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| 作者单位： | 1. 中国农业科学院哈尔滨兽医研究所,兽医物技术国家重点实验室/农业部动物流感重点开放实验室,黑龙江,哈尔滨,150001;东北农业大学,动物医学学院,黑龙江,哈尔滨,150030
2. 中国农业科学院哈尔滨兽医研究所,兽医物技术国家重点实验室/农业部动物流感重点开放实验室,黑龙江,哈尔滨,150001 |
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| 摘 要： | 采用纯化的H3N2亚型猪流感病毒(SIV)尿囊液作为免疫原免疫6～8周龄Balb/c小鼠,取免疫小鼠脾细胞与骨髓瘤细胞(SP2/0)融合,用间接ELISA方法筛选分泌抗SIV-H3N2的阳性细胞株,经克隆获得7株亲和力较高的杂交瘤细胞株,分别命名为1C9、2C5、2F10、3D3、4E8、5C7、5D12,用其制备的腹水ELISA效价可达1×106。通过抗体亚型测定,间接免疫荧光试验及免疫印迹试验分析鉴定,该7株单抗均为抗H3N2亚型SIV的特异性单克隆抗体,而且与其他亚型猪流感病毒、猪细小病毒、猪繁殖与呼吸综合征病毒、猪圆环病毒和猪瘟病毒等均无交叉反应,为H3N2亚型SIV的鉴别诊断奠定了基础。
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| 关 键 词： | 猪流感病毒 单克隆抗体 ELISA |
Preparation and identification of monoclonal antibodies against H3N2 subtype swine influenza virus |
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| KONG Wei,QIAO Chuan-ling,CHEN Yan,YANG Huan-liang,ZHAN Xiao-guo,XIN Xiao-guang,CHEN Hua-lan.Preparation and identification of monoclonal antibodies against H3N2 subtype swine influenza virus[J].Animal Husbandry & Veterinary Medicine,2010,42(9). |
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| Authors: | KONG Wei QIAO Chuan-ling CHEN Yan YANG Huan-liang ZHAN Xiao-guo XIN Xiao-guang CHEN Hua-lan |
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| Institution: | KONG Wei1,2,QIAO Chuan-ling1,CHEN Yan1,YANG Huan-liang1,ZHAN Xiao-guo1,XIN Xiao-guang1,CHEN Hua-lan1(1.Animal Influenza Laboratory of the Ministry of Agriculture,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China,2.College of Veterinary Medicine,Northeast Agriculture University,Harbin 150030,China) |
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| Abstract: | Seven monoclonal antibodies(McAbs) against H3N2 subtype swine influenza virus(SIV) were prepared and identified by fusing SP2/0 cell with splenic cells of Balb/c mice that had been immunized with the purified virus antigen.Positive hybridoma cell strains 1C9,2C5,2F10,3D3,4E8,5C7 and 5D12 were determined by ELISA,which showed the highest ELISA titers of 1×106 in ascites.These McAbs were specific to SIV H3N2 by subtype identification,indirect immunofluoescence assay and western-blot.There were no cross reaction with other subtypes of SIV,porcine parvovirus(PPV),porcine reproductive and respiratory syndrome virus(PRRSV),porcine circovirus(PCV) and classical swine fever virus(CSFV).The McAbs are sensitive and specific,and suitable to be applied in identification of H3N2 subtype SIV. |
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| Keywords: | ELISA |
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