Production of Monoclonal Antibodies against Vibrio parahaemolyticus and Their Application to Immunoassay Using Magnetic Nanoparticle

Introduction: Vibrio parahaemolyticus are Gram negative halophilic marine bacteria occurring naturally in the estuarine environment and are one of the major reasons developing acute gastroenteritis.

Purpose: The objectives of this study are to produce monoclonal antibodies (MAbs) against V. parahaemolyticus and to develop immuno-selective filtration for the rapid detection of V. parahaemolyticus.

Methods: To produce monoclonal antibodies against V. parahaemolyticus, heat killed V. parahaemolyticus (HKVP) and formalin killed V. parahaemolyticus (FKVP) were prepared. The MAb for V. parahaemolyticus was produced from cell fusion and cloning. MAb was conjugated to 74 nm magnetic nanoparticles to develop the immuno-selective filtration.

Results: A MAb was produced from HKVP 4H9-9, 16 hybridoma cell. HKVP 4H9-9 MAb was conjugated to 74 nm magnetic nanoparticles to develop the immuno-selective filtration. The detection limit of immuno-selective filtration for V. parahaemolyticus was 10¹ cell/ml. The immuno-selective filtration was specific to V. parahaemolyticus. Cross-reactivity to V. alginolyticus and S. aureus was.

Significance: These results strongly support that the application to immuno-selective filtration could be used as rapid, simple and effective methods.