| 摘要: |
| 【目的】制备抗鸭副粘病毒(DPMV)的单克隆抗体，为鸭副粘病毒病免疫学诊断方法的建立奠定基础。【方法】采用差速和蔗糖不连续密度梯度离心法提纯DPMV，免疫BALB/c 小鼠，取脾细胞与骨髓瘤细胞株SP2/0融合，采用间接 ELISA、间接免疫荧光试验（IFA）和血凝抑制（HI）试验等方法筛选阳性杂交瘤细胞株，并测定其生物学特性。【结果】筛选到7株能稳定分泌抗DPMV单克隆抗体的杂交瘤细胞株，分别命名为J28、F4、B3、E39、E41、A12和A30。这7株单克隆抗体都具有 ELISA和1FA特性，其中J28和F4还具有HI和中和特性。免疫球蛋白亚类测定结果显示，J28和F4均为IgG1，其他5株均为IgM。相加ELISA试验结果显示，单抗B3，J28与F4，A30与E39/E41/A12分别识别3个不同的抗原位点。特异性测定结果显示，7株单克隆抗体与雏番鸭细小病毒(MPV)、鹅细小病毒(GPV)、鸭肝炎病毒(DHV)、番鸭呼肠孤病毒(MDRV)及正常细胞均无交叉反应。【结论】筛选出了7株具有ELISA、1FA等特性且分泌抗DPMV单克隆抗体的杂交瘤细胞株，并明确了各株单克隆抗体的特性，为鸭副粘病毒病的免疫治疗和临床诊断奠定了基础。 |
| 关键词: 鸭源副粘病毒；单克隆抗体；ELISA IFA HI |
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| 基金项目:福建省农业科学院动物传染病创新团队资助项目（STIF-Y02） |
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| Preparation and characterization of monoclonal antibodies against duck paramyxovirus infection |
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| Abstract: |
| 【Objective】The paper was to obtain monoclona1 antibodies against duck paramyxovirus for further application to clinic test.【Method】Duck paramyxovirus was propagated in chicken embryo and purified by differential centrifugation and discontinuous sucrose density gradient centrifugation.The spleen cells of Balb/c mice immunized with purified viral antigen were fused with SP2/0 myeloma cells.Hybridoma cell lines secreting monoclona1 antibodies against duck paramyxovirus were screened by using indirect ELISA,IFA and HI.The specificities of these monoclonal antibodies were determined by immunoglobulin G subclass and ELISA.【Result】Seven monclonal antibodies against duck paramyxovirus were obtained using hybrodoma technique.They were designated as J28,F4,B3,E39,E41,A12 and A30.These McAbs all had ELISA and IFA activity,and two of them J28 and F4 had HI and neutralizing activity.The isotyping analysis showed that both J28 and F4 belonged to IgG1,and others belonged to IgM.It was found by ELISA additivity test that the seven McAbs recognized three different antigenic sites,in which,the epitopes of J28 and F4,E39,E41,A12 and A30 were very close,respectively,and the epitope of B3 was solely.Result of specificity test indicated that all these monoclonal antibodies were specific to duck paramyxovirus (DPMV),and did not react with Muscovy duck parvovirus (MPV),Goose parvovirus(GPV),Duck hepatitis virus (DHV),and Muscovy duck reovirus(MDRV).【Conclusion】The DPMV McAbs were successfully prepared,which could provide a good foundation for further research on the immune therapy and clinical diagnosis. |
| Key words: duck paramyxovirus monoclonal antibody ELISA 1FA HI |
