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对虾白斑综合症病毒双抗体夹心ELISA方法的建立 被引量:4

Development of Double Antibody Sandwich ELISA for Detection of White Spot Syndrome Virus
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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。 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.
出处 《中国动物检疫》 CAS 2012年第4期41-43,共3页 China Animal Health Inspection
关键词 对虾白斑综合症病毒 VP28蛋白 单克隆抗体 ELISA white spot syndrome vires VP28 protein monoclonal antibody double antibody sandwich ELISA
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参考文献5

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  • 2Sophie St-Hilaire,Carl S Ribble,Craig Stephen,Eric Anderson,Gael Kurath,Michael L Kent.Epidemiological investigation of infectious hematopoietic necrosis virus in salt water net-pen reared Atlantic salmon in British Columbia, Canada[J].Aquaculture.2002(1)
  • 3KOverturf,SLaPatra,MPowell.Real‐time PCR for the detection and quantitative analysis of IHNV in salmonids[J].Journal of Fish Diseases.2001(6)
  • 4Marta Alonso,Sylvia Rodr??guez,Sara I Pérez Prieto.Nested PCR improves detection of infectious hematopoietic necrosis virus in cells coinfected with infectious pancreatic necrosis virus[J].Journal of Virological Methods.1999(1)
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