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Detection methods of Cyprinid herpesvirus 2 infection in silver crucian carp ( Carassius auratus gibelio ) via a pORF 72 monoclonal antibody
Author(s) -
Kong S Y,
Jiang Y S,
Wang Q,
Lu J F,
Xu D,
Lu L Q
Publication year - 2017
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/jfd.12648
Subject(s) - crucian carp , biology , carassius , monoclonal antibody , carassius auratus , carp , virology , zoology , antibody , fishery , microbiology and biotechnology , fish <actinopterygii> , immunology
Cyprinid herpesvirus 2 (Cy HV ‐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 Cy HV ‐2 in clinical samples using immunological methods, recombinant ORF 72 protein ( pORF 72), encoded by the Cy HV ‐2 ORF 72 gene, was used as a capture antigen to identify blood and tissues infected with Cy HV ‐2. First, ORF 72 gene was amplified from the Cy HV ‐2 genome and cloned into a PGEX ‐4t‐3 expression vector to produce pORF 72 in Escherichia coli . The purified pORF 72 was used as an immunogen to prepare monoclonal antibodies. The Western blotting assays revealed that the monoclonal antibody could specifically identify the pORF 72. Furthermore, an immunohistochemical protocol and a blood smear method were established to detect Cy HV ‐2 in carps. The results indicate that the monoclonal antibody against pORF 72 could be utilized as an effective detection tool for haematopoietic necrosis disease in Carassius auratus gibelio .