| 摘要: |
| 【目的】 制备猪圆环病毒Ⅱ型（PCV2 ）Cap蛋白单克隆抗体，并对其进行鉴定和应用。【方法】 以超离纯化的猪圆环病毒Ⅱ型作为免疫原，按常规方法免疫BALB/c小鼠，取其脾细胞与骨髓瘤细胞（SP2/0）融合，经间接ELISA筛选获得了2株分泌PCV2-Cap蛋白单克隆抗体的杂交瘤细胞，分别命名为8-60和10-48，对其生物学特性进行鉴定，并将其作为一抗，建立了间接免疫荧光诊断方法。【结果】 8-60和10-48的染色体数平均为98~105对，间接ELISA检测其细胞培养上清液效价分别为1∶16和1∶32，小鼠腹水效价分别为1∶3×212和1∶3×213，抗体的免疫球蛋白亚型均为IgM。间接免疫荧光结果显示，8-60和10-48能与PCV2发生反应；利用这种特性，建立了能诊断PCV2病原的间接免疫荧光诊断方法，实际应用结果表明，其诊断结果与PCR方法诊断结果基本一致，阳性和阴性的符合率分别为93.75%和100%。【结论】 以制备的猪圆环病毒Ⅱ型（PCV2）Cap蛋白单克隆抗体为一抗，建立了间接免疫荧光诊断方法。 |
| 关键词: 猪圆环病毒Ⅱ型 单克隆抗体 Cap蛋白 ELISA方法 间接免疫荧光 |
| DOI： |
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| 基金项目:福建省科技计划重点项目（2006N0026） |
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| Preparation and application of monoclonal antibody against Porcine circovirus Ⅱ |
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| Abstract: |
| 【Objective】The monoclonal antibodies against Porcine circovirus Ⅱ (PCV2) Cap were prepared,identified and applied.【Method】 BALB/c mice were regularly immunized with purified PCV2 by ultracentrifugation.Spleen cells were collected and infused with SP2/0 cell.Two hybridoma cell lines against PCV2 Cap named 8-60 and 10-48 were obtained by indirect enzyme linked immunosorbent assay (ELISA) method.And we identified the biological character of the two monoclonal antibodies,and established indirect immunofluorescent method by these antibodies as antigen.【Result】 The average number of chromosome of the hybridoma cell lines was 98-105.The ELISA titer for Cap antibodies in culture supernant was 1∶16 and 1∶32 ；The ELISA titer in ascites was 1∶3×212 and 1∶3×212.The Ig subtype of these antibodies was IgM.Immunofluorescence assay(IFA）test showed that the two monoclonal antibodies could react with PCV2 virus.Applying the propertity,an indirect immunofluorescent method for the PCV2 antigen was established,and the actual appliction result showed the diagnosis was nearly in accordance with the result of PCR,and the positive according rate was 93.75%,the negative according rate 100%.【Conclusion】 We successfully established the indirect immunofluorescent method by the monoclonal antibodies prepared as the first antibodies. |
| Key words: Porcine circovirus Ⅱ monoclonal antibody Cap protein ELISA method immunofluorescence assay (IFA) |
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