| 对虾白斑综合症病毒双抗体夹心ELISA方法的建立 |
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| 引用本文： | 郑晓聪,刘荭,张朋,何俊强,史秀杰,贾鹏,兰文升.对虾白斑综合症病毒双抗体夹心ELISA方法的建立[J].动物检疫,2012(4):41-43. |
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| 作者姓名： | 郑晓聪 刘荭 张朋 何俊强 史秀杰 贾鹏 兰文升 |
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| 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518045 [2]华中农业大学水产学院,湖北武汉430070 |
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| 摘 要： | 用将纯化的对虾白斑综合症病毒VP28蛋白免疫BALB/c小鼠,分离免疫鼠脾细胞,与SP2/0细胞融合,经间接ELISA筛选,得到了2株（2G9,3E5）可以稳定分泌抗对虾白斑综合症病毒特异性单抗的杂交瘤细胞株。用纯化的VP28蛋白免疫家兔,按常规方法制备多抗。选用单抗（3E5）包被ELISA板,用兔多抗作为捕获抗体,建立了对虾白斑综合症病毒的双抗体夹心ELISA检测方法。该方法具有良好的特异性和敏感性,为对虾白斑综合症病毒的检测提供了有效工具。
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| 关 键 词： | 对虾白斑综合症病毒 VP28蛋白 单克隆抗体 ELISA |
Development of Double Antibody Sandwich ELISA for Detection of White Spot Syndrome Virus |
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| Authors: | Zheng Xiaocong Liu Hong Zhang Peng He Junqiang Shi Xiujie Jia Peng Lan Wensheng |
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| Institution: | 1. Technic Centre of Animal and Plant Inspection and Quarantine, Shenzhen Entry ＆ Exit Inspection and Quarantine Bureau, Shenzhen 518045, China; 2. College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China) |
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| Abstract: | SPF BALB/c mice were immunized with purified VP28 protein of white spot syndrome virus （WSSV） emulsified with Fretmd＇s adjuvant. The spleen cell isolated from vaccinated mice were fused with SP2/0 cells. Two hybridoma of 2G9 and 3 E5, screened using ELISA, could secrete stably specific antibodies to VP28 protein of WSSV A double antibody sandwich ELISA （DAS-ELISA） for detection of WSSV was established based on monoclonal antibody （3E5） and rabbit-anti-VP28 polyclonal antibody. The result showed that the DAS-ELISA had good specificity and sensitivity, indicating that the method was suitable for rapid detection and epidemiological investigation of WSSV infection in shrimps. |
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| Keywords: | white spot syndrome vires VP28 protein monoclonal antibody double antibody sandwich ELISA |
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