Date of Award

1989

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

First Advisor

R. J. Siebeling

Abstract

Current schemes used to serotype Vibrio vulnificus are based on direct bacterial agglutination using polyclonal rabbit antisera. During the course of serotyping Louisiana V. vulnificus isolates we failed to observe a reproducible agglutination profile for these isolates. The major obstacle in producing serovar specific sera has been the failure to produce high titered anti-LPS sera in rabbits. Anti-LPS serum from rabbits immunized with formalin-killed, heat-killed, or acetone dried whole cell vaccines and LPS-carrier conjugates uniformly produced titers $\leq 320$. Separation of anti-LPS activity from whole sera by affinity chromatography also yielded low titered antisera. A typing system predicated on the detection of LPS-associated O antigens using monoclonal antibodies (Mab) is proposed for V. vulnificus. Hybridomas have been established which produce antibodies that react with LPS purified from V. vulnificus, V. damsela, and V. hollisae. Vibrio vulnificus shows considerable heterogeneity within the genus. Twenty-four Mabs produced to date, react with 10 of 16 (61%) of V. vulnificus strains tested by LPS ELISA. Monoclonal antibodies react more strongly with the translucent than the opaque phenotype of V. vulnificus using whole cell ELISA. Monoclonal antibodies reactive by LPS ELISA with V. damsela and V. hollisae LPS are non-reactive with LPS prepared from V. vulnificus.

Pages

100

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