对虾白斑综合症病毒双抗体夹心ELISA方法的建立  

Development of Double Antibody Sandwich ELISA for Detection of White Spot Syndrome Virus

作　　者：郑晓聪[1] 刘荭[1] 张朋[2] 何俊强[1] 史秀杰 贾鹏[1] 兰文升[1] 

机构地区：[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

出　　处：《中国动物检疫》2012年第4期41-43,共3页China Animal Health Inspection

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。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.

关 键 词：对虾白斑综合症病毒 VP28蛋白 单克隆抗体 ELISA 

分 类 号：S945.46[农业科学—水产养殖]