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禽脑脊髓炎病毒单克隆抗体的制备与初步应用
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徐建生，唐波，成大荣，董国雄，吕玲，曹军
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（扬州大学 兽医学院，江苏 扬州225009）
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摘要：
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利用提纯的禽脑脊髓炎病毒（AEV）Van Roekel 株作为免疫原，免疫6周龄BALB/c小鼠，采取其脾细胞与SP2/0骨髓瘤细胞融合，用间接ELISA法筛选，间接免疫荧光法（IFA）和免疫组织化学法鉴定，经3次亚克隆得到了稳定分泌抗AEV单克隆抗体的杂交瘤细胞株F11和G2，制备了腹水，并利用该单克隆抗体初步建立了DotELISA、间接ELISA和IFA等特异性检测AEV抗原的方法。 |
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关键词：
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禽脑脊髓炎病毒；单克隆抗体；斑点酶联免疫吸附法 |
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中图分类号：S 852.659.6: Q 813.2 文献标识码：
A 文章编号：1673-4696(2006)02-0089-04 |
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Preparation and preliminary application of monoclonal
antibody to avian encephalomyelitis virus |
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XU Jiansheng, TANG Bo, CHENG Darong, DONG Guoxiong,
Lu Ling ,CAO Jun |
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(College of Veterinary Medicine, Yangzhou University, Yangzhou 225009,China) |
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Abstract: 6week old BALB/c mice were immunized with purified avian encephalomyelitis viruses at two weeks interval. Mice were killed and their spleen cells fused into SP2/0 myeloma cells on day 3 after third immunization. IndirectELISA, indirect immunofluorescent assay and immunohistochemistry were employed to select and characterize the fused cells secreting antibody to AEV. As a result, two hybridoma cell lines steadily secreting antibody to AEV, named F11and G2, were obtained after subcloning for three times. Ascites were prepared with the hybridoma and indirect fluorescent assay, indirect ELISA and DotELISA were developed with the monoclonal antibody for the detection of AEV. |
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Key words: avian encephalomyelitis virus; monoclonal antibody; DotELISA |
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