Development and Application of Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibody against a Hemolysin (Vp-TRH) of Vibrio parahemolyticus

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Abstract
Although it has been well established that Kanagawa phenomenon-positive Vibrio parahaemolyticus is a human enteropathogen, the Kanagawa phenomenon-negative one has been considered to be probably not pathogenic. We have found, however, an outbreak of gastroenteritis due to Kanagawa phenomenon-negative V. parahaemolyticus which produces a new toxin (Vp-TRH) resembling to Vp-TDH, a responsible toxin of Kanagawa phenomenon. In this study, we developed monoclonal antibodies against Vp-TRH which were used for development of an enzyme-linked immunosorbent assay (ELISA) for specifically detecting Vp-TRH. The ELISA was applied for analysis of production of Vp-TRH by various isolates of V. parahaemolyticus and we found that Vp-TRH-producing strains were derived mostly from human diarrheal stool, and not from the environment or sea foods. The results of the rabbit ileal loop test showed that Vp-TRH-producing (Kanagawa phenomenon-negative) strains, as well as Vp-TDH-producing (Kanagawa phenomenon-positive) strains could induce fluid accumulation. These results indicate the possibility that Vp-TRH-producing Kanagawa phenomenon-negative V. parahaemolyticus is a human enteropathogen.

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