Production of monoclonal antibodiesfor detection of Vibrio harveyi

Diseases of Aquatic Organisms

Wannipa Phianphak Sirirat Rengpipat Sombat Rukpratanporn Siwaporn Longyant Parin Chaivisuthangkura Weerawan Sithigorngul Paisarn Sithigorngul¹ 

关键词: Dot blot;    Immunohistochemistry;    Monoclonal antibody;    Vibrio harveyi;    Penaeus monodon;    Western blot;   

DOI  :  10.3354/dao063161

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

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 totheir 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 ofthe 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 studydemonstrates 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.

【 授权许可】

Unknown