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Diseases of Aquatic Organisms

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DAO 42:27-34 (2000)  -  doi:10.3354/dao042027

Development of a monoclonal antibody specific to yellow head virus (YHV) from Penaeus monodon

Paisarn Sithigorngul1,*, Phudhi Chauychuwong2, Weerawan Sithigorngul1, Siwaporn Longyant1, Parin Chaivisuthangkura1, Piamsak Menasveta2-4

1Department of Biology, Faculty of Science, Srinakharinwirot University, Bangkok 10110, Thailand
2Marine Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology, Ministry of Science, Technology and Environment, Bangkok 10400, Thailand
3Aquatic Resources Research Institute, and
4Department of Marine Science, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand

ABSTRACT: A monoclonal antibody specific to yellow head virus (YHV) was produced from a mouse immunized with gill extracts prepared from laboratory-reared Penaeus monodon dually infected with YHV and white spot syndrome virus (WSSV). One clone designated V3-2B specifically bound to native and SDS-treated viral specific antigens. Immunocytochemical studies of infected gills revealed viral specific immunoreactivities in the cytoplasm of gill tissue and in haemocytes. No antibody binding was observed in gills from non-infected shrimp. In addition, immunocytochemical examination of tissues from shrimp experimentally infected with YHV gave a positive reaction, while tissues from uninfected control shrimp or shrimp experimentally infected with WSSV did not. Western blot analysis indicated that the antibody reacted with a protein of approximately 135 kD that was present only in shrimp infected with YHV. In dot-blot indirect immunoperoxidase assays, the antibody was able to detect viral associated antigen in diluted haemolymph up to 1:50 dilution and in an ammonium sulfate precipitate of haemolymph up to 1:1000 dilution. The results suggested that this antibody might be useful for development of effective diagnostic techniques for both heavy and mild YHV infections in shrimp.

KEY WORDS: Immunocytochemistry · Monoclonal antibodies · Penaeus monodon · Yellow head virus (YHV)

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