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| 抗H9N2 AIV HA单克隆抗体的制备及其在胶体金层析检测中的初步应用 | |
| 引用本文： | 詹爱军,王新卫,陈枝楠,卢体康,花群义.抗H9N2 AIV HA单克隆抗体的制备及其在胶体金层析检测中的初步应用[J].河南农业科学,2008(9). |
| 作者姓名： | 詹爱军 王新卫 陈枝楠 卢体康 花群义 |
| 作者单位： | 1. 深圳出入境检验检疫局,动植物检验检疫技术中心,广东,深圳,518010;扬州大学,兽医学院,江苏,扬州,225009 2. 河南农业大学,牧医工程学院,河南,郑州,450002 3. 深圳出入境检验检疫局,动植物检验检疫技术中心,广东,深圳,518010 |
| 摘 要： | 用基因疫苗pcDNA-H9和纯化H9N2亚型AIV免疫Balb/C小鼠,细胞融合后用HI和ELISA方法筛选出5株抗H9亚型AIV HA的特异性单克隆抗体H9-01(2E6),H9-02(3B1),H9-03(3H11),H9-04(1A4),H9-05(2G9)。各株单抗亚型测定的结果为H9-01,H9-02和H9-05均为IgG2b,H9-03为IgG1,H9-04为IgG2a,轻链的亚型均为kappa链。经特异性和与同型病毒的反应谱检测,它们均是HA特异性单克隆抗体。用制备的抗H9亚型AIV HA的单克隆抗体做成胶体金层析检测试纸条,可以检测到200个EID50的H9亚型AIV,为监测该亚型AIV试剂盒的进一步商品化奠定了基础。 |
| 关 键 词： | H9N2亚型AIV 单克隆抗体 胶体金层析检测 |
Preparation of Monoclonal Antibodies against HA of H9N2 AIV and Its Preliminary Application in Colloidal Gold Immunochromatographic Assay |
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| ZHAN Ai-jun,WANG Xin-wei,CHEN Zhi-nan,LU Ti-kang,HUA Qun-yi.Preparation of Monoclonal Antibodies against HA of H9N2 AIV and Its Preliminary Application in Colloidal Gold Immunochromatographic Assay[J].Journal of Henan Agricultural Sciences,2008(9). | |
| Authors: | ZHAN Ai-jun WANG Xin-wei CHEN Zhi-nan LU Ti-kang HUA Qun-yi |
| Abstract: | Balb/C mice were immunized with pcDNA-H9 gene vaccine or purified H9N2 subtype AIV.After fusion of the splenocytes obtained from the immunized mice with myeloma cells,five hybridoma cell lines including H9-01(2E6),H9-02(3B1),H9-03(3H11),H9-04(1A4) and H9-05(2G9) which persistently secrete specific monoclonal antibodies against H9 subtype AIV,were finally selected by HI and ELISA.Testing for the subgroup of the monoclonal antibodies revealed that H9-01,H9-02 and H9-05 belonged to the subgroup IgG2b,H9-03 to IgG1,H9-04 to IgG2a,respectively.All of their light chains are of the kappa subtype.Specificity and homo-logous virus reaction spectrum testing showed that they were all HA-specific monoclonal antibodies.A colloidal gold immunochromatographic assay was developed for the detection of H9 AIV by using these monoclonal antibodies as diagnostic agent,and 200 EID50 H9 AIV could be de-tected by this method.Our results provided useful data for further developing the diagnostic kit to detect the virus in practical application. |
| Keywords: | H9N2 subtype avian influenza virus Monoclonal antibody Colloidal gold immunochromatographic assay |
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