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Detection methods of Cyprinid herpesvirus 2 infection in silver crucian carp (Carassius auratus gibelio) via a pORF72 monoclonal antibody

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文献类型：外文期刊

作者： Kong, S. Y. ¹ ; Jiang, Y. S. ¹ ; Wang, Q. ² ; Lu, J. F. ¹ ; Xu, D. ¹ ; Lu, L. Q. ¹ ;

作者机构： 1.Shanghai Ocean Univ, Minist Aquaculture, Key Lab Aquat Genet Resources, Natl Pathogen Collect Ctr Aquat Anim, Shanghai, Peoples R China

2.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Guangzhou, Guangdong, Peoples R China

关键词： blood smear;Carassius auratus gibelio;Cyprinid herpesvirus 2;monoclonal antibody;pORF72;immunohistochemistry

期刊名称：JOURNAL OF FISH DISEASES （影响因子：2.767；五年影响因子：2.689 ）

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收录情况： SCI

摘要： Cyprinid herpesvirus 2 (CyHV-2) is the main pathogen responsible for causing haematopoietic necrosis disease in Carassius auratus gibelio. Although many nucleic acid-based diagnostic methods have been applied, no stable and sensitive immunological diagnostic approaches have been reported. In this study, to detect CyHV-2 in clinical samples using immunological methods, recombinant ORF72 protein (pORF72), encoded by the CyHV-2 ORF72 gene, was used as a capture antigen to identify blood and tissues infected with CyHV-2. First, ORF72 gene was amplified from the CyHV-2 genome and cloned into a PGEX-4t-3 expression vector to produce pORF72 in Escherichia coli. The purified pORF72 was used as an immunogen to prepare monoclonal antibodies. The Western blotting assays revealed that the monoclonal antibody could specifically identify the pORF72. Furthermore, an immunohistochemical protocol and a blood smear method were established to detect CyHV-2 in carps. The results indicate that the monoclonal antibody against pORF72 could be utilized as an effective detection tool for haematopoietic necrosis disease in Carassius auratus gibelio.

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