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Diseases of Aquatic Organisms

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DAO 63:161-168 (2005)  -  doi:10.3354/dao063161

Production of monoclonal antibodiesfor detection of Vibrio harveyi

Wannipa Phianphak1, Sirirat Rengpipat1, Sombat Rukpratanporn2, Siwaporn Longyant3, Parin Chaivisuthangkura3, Weerawan Sithigorngul3, Paisarn Sithigorngul3,*

1Biotechnology Program, and 2Marine Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology, Ministry of Science and Technology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand
3Department of Biology, Faculty of Science, Sukhumvit 23, Srinakharinwirot University, Bangkok 10110, Thailand
*Corresponding author. Email:

ABSTRACT: Monoclonal antibodies (MAbs) against Vibrio harveyi were produced from mice immunized with heat-killed and SDS-mercaptoethanol-treated highly virulent V. harveyi 639. Fifteen MAbs were selected and sorted into 6 groups according to their specificity to various proteins of apparent molecular weight ranging from 8 to 49 kDa. Some antibodies were used for detection of V. harveyi at concentrations as low as 104 CFU ml–1 using immunodot blots. Most of the selected MAbs did not show cross-reactivity to other Vibrio species and other Gram-negative bacteria tested. Only 1 MAb (VH39-4E) showed slight cross-reactivity to Aeromonas hydrophila. Another MAb (VH24-8H) bound lightly to V. harveyi 1526 but strongly to V. harveyi 639, allowing rapid differentiation. Two of the MAb groups were used to localize V. harveyi in tissues of infected black tiger shrimp Penaeus monodon by immunohistochemistry. This study demonstrates the versatility of a highly specific immunological tool for the detection of V. harveyi in aquaculture and opens the way for further development of convenient test kits.

KEY WORDS: Dot blot · Immunohistochemistry · Monoclonal antibody · Vibrio harveyi · Penaeus monodon · Western blot

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