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Production of monoclonal antibodies against thermostable direct hemolysin of Vibrio parahaemolyticus and application of the antibodies for enzyme-linked immunosorbent assay

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Abstract

A total nine hybridoma cell lines that produced monoclonal antibodies against thermostable direct hemolysin (Vp-TDH), a possible pathogenic toxin, of Kanagawa phenomenon-positive Vibrio parahaemolyticus was isolated and characterized. These monoclonal antibodies (mAbs) were divided into a minimum of five different specificity groups, including mAbs specific to Vp-TDH and common to Vp-TDH and Vp-TRH, a Vp-TDH-related hemolysin produced by Kanagawa phenomenon-negative V. parahaemolyticus. An enzyme-linked immunosorbent assay (ELISA) using mAb-1-D, a mAb specific for Vp-TDH, was developed for specific detection of Vp-TDH. On the other hand, the ELISA using mAb-9-D, and mAb common to both Vp-TDH and Vp-TRH, could be used for detection of both Vp-TDH and Vp-TRH. Thus, by combining these two ELISAs differential detection of Vp-TDH and Vp-TRH can be performed. Hence, the two ELISAs were applied for various strains of V. parahaemolyticus and it was found that most Kanagawa phenomenon-positive and -negative clinical isolates produced Vp-TDH and Vp-TRH, respectively, but all environmental strains, that were Kanagawa phenomenon-negative, produced neither toxin.

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References

  1. Chan D, Chung JK, Tak Y, Seol SY (1975) Nature of the Kanagawa phenomenon of Vibrio parahaemolyticus. Infect Immun 12:81–87

    Google Scholar 

  2. Davis BJ (1964) Disc electrophoresis. II. Method and application to human serum proteins. Ann NY Acad Sci 121:404–427

    Google Scholar 

  3. Galfre G, Howe SC, Milstein C, Butcher GW, Howard JC (1977) Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature 266:530–552

    Google Scholar 

  4. Holmes R, Twiddy EM (1983) Characterization of monoclonal antibodies that react with unique and cross-reacting determinants of cholera enterotoxin and its subunits. Infect Immun 42:914–923

    Google Scholar 

  5. Honda T, Goshima K, Takeda Y, Sugino Y, Miwatani T (1976) Demonstration of the cardiotoxicity of the thermostable direct hemolysin (lethal toxin) produced by V. parahaemolyticus. Infect Immun 13:163–171

    Google Scholar 

  6. Honda T, Taga S, Takeda T, Hashibunan MA, Takeda Y, Miwatani T (1976) Identification of lethal toxin with the thermostable direct hemolysin produced by Vibrio parahaemolyticus and some physicochemical properties of the purified toxin. Infect immun 13:133–139

    Google Scholar 

  7. Honda T, Chearkul S, Takeda Y, Miwatani T (1980) Immunological methods for detection of Kanagawa phenomenon of Vibrio parahaemolyticus. J Clin Microbiol 11:600–603

    Google Scholar 

  8. Honda T, Sornchai C, Takeda Y, Miwatani T (1982) Immunological detection of the Kanagawa phenomenon of Vibrio parahaemolyticus on modified selective media. J Clin Microbiol 16:734–736

    Google Scholar 

  9. Honda T, Yoh M, Kongmuang U, Miwatani T (1985) Enzyme-linked immunosorbent assays for detection of thermostable direct hemolysin of Vibrio parahaemolyticus. J Clin Microbiol 22:383–386

    Google Scholar 

  10. Honda T, Ni Y, Miwatani T (1988) Purification and characterization of a hemolysin produced by a clinical isolate of Kanagawa phenomenon-negative Vibrio parahaemolyticus and related to the thermostable direct hemolysin. Infect Immun 56:961–965

    Google Scholar 

  11. Honda T, Ni Y, Miwatani T (1989) Purification of a TDH-related hemolysin produced by a Kanagawa phenomenon negative clinical isolate of Vibrio parahaemolyticus 06:K46. FEMS Microbiol Lett 57:241–246

    Google Scholar 

  12. Janda JM, Powers C, Bryant RG, Abbott SL (1988) Current perspectives on the epidemiology and pathogenesis of clinically significant Vibrio spp. Clin Microbiol Rev 1:245–267

    Google Scholar 

  13. Joseph SW, Colwell RP, Kaper JB (1982) Vibrio parahaemolyticus and related halophilic vibrios. Crit Rev Microbiol 10:77–124

    Google Scholar 

  14. McKearn TJ (1980) Fusion of cells in an adherent monolayer. In: Kennett RH, McKearn TJ, Bechtol KB (ed) Monoclonal antibodies, hybridomas: a new dimension in biological analyses. Plenum Press, New York, pp 368–369

    Google Scholar 

  15. Miwatani T, Takeda Y (1976) Vibrio parahaemolyticus—a causative bacterium of food poisoning. Saikon, Tokyo, pp 1–109

    Google Scholar 

  16. Miyake M, Honda T, Miwatani T (1989) Effects of divalent cations and saccharides on Vibrio metschnikovii cytolysin-induced hemolysis of rabbit erythrocytes. Infect Immun 57:158–163

    Google Scholar 

  17. Miyamoto Y, Kato T, Obara Y, Akiyama S, Takizawa K, Yamai S (1969) In vitro hemolytic characteristics of Vibrio parahaemolyticus: its close correlation with human pathogenicity. J Bacteriol 100:1147–1149

    Google Scholar 

  18. Miyamoto Y, Obara Y, Nikkawa T, Yamai S, Kato T, Yamada Y, Ohashi M (1980) Simplified purification and biophysicochemical characteristics of Kanagawa-associated hemolysin of V. parahaemolyticus. Infect Immun 23:567–576

    Google Scholar 

  19. Morris JG Jr, Blake RE (1985) Cholera and other vibrioses in the United States. N Engl J Med 312:343–350

    Google Scholar 

  20. Nishibuchi M, Kaper JB (1985) Nucleotide sequence of the thermostable direct hemolysin gene of Vibrio parahaemolyticus. J Bacteriol 162:558–564

    Google Scholar 

  21. Oku Y, Uesaka Y, Hirayama T, Takeda Y (1988) Development of a highly sensitive bead-ELISA to detect bacterial toxins. Microbiol Immunol 32:807–816

    Google Scholar 

  22. Sakazaki R, Tamura K, Kato T, Obara Y, Yamai S, Hobo K (1968) Studies on the enteropathogenic, facultatively halophilic bacteria, Vibrio parahaemolyticus. III. Enteropathogenicity. Jpn J Med Sci Biol 21:325–331

    Google Scholar 

  23. Sakurai J, Honda T, Jinguji Y, Arita M, Miwatani T (1976) Cytotoxic effect of the thermostable direct hemolysin produced by V. parahaemolyticus on FL cells. Infect Immun 13:876–883

    Google Scholar 

  24. Takeda Y, Taga S, Miwatani T (1978) Evidence that thermostable direct hemolysin of V. parahaemolyticus is composed of two subunits. FEMS Microbiol Lett 4:271–274

    Google Scholar 

  25. Toda H, Sakiyama F, Yoh M, Honda T, Miwatani T (1985) Chemical modification of thermostable direct hemolysin of V. parahaemolyticus. Jpn J Med Sci Biol 38:264–265

    Google Scholar 

  26. Tsunasawa S, Sugihara A, Masaki T, Sakiyama F, Takeda Y, Miwatani T, Narita K (1987) Amino acid sequence of thermostable direct hemolysin produced by V. parahaemolyticus. J Biochem 101:11–121

    Google Scholar 

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Authors and Affiliations

  1. Research Institute for Microbial Diseases, Osaka University, Yamadaoka, Suita, 565, Osaka, Japan

    T. Honda, Y. Ni, M. Yoh & T. Miwatani

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  1. T. Honda
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  2. Y. Ni
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  3. M. Yoh
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  4. T. Miwatani
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Supported by a Grant-in-Aid for Scientific Research and a Grant-in-Aid for Developmental Scientific Research from the Ministry of Education, Science, and Culture of Japan

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Honda, T., Ni, Y., Yoh, M. et al. Production of monoclonal antibodies against thermostable direct hemolysin of Vibrio parahaemolyticus and application of the antibodies for enzyme-linked immunosorbent assay. Med Microbiol Immunol 178, 245–253 (1989). https://doi.org/10.1007/BF00191059

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  • DOI: https://doi.org/10.1007/BF00191059

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