Corynebacterium pseudotuberculosis is a gram-positive bacterium that causes abscesses in small ruminants and inhabits human skin and mucous membranes. A number of coryneform bacteria have been isolated from patients with endocarditis and bone marrow transplant. Currently, the radioimmunoassay (example, synergistic hemolysin) tests used for the diagnosis of C. pseudotuberculosis are insensitive and non-specific. Therefore, we propose the use of modern day molecular biological technology for the development of diagnostic markers for C. pseudotuberculosis. The cell surface antigens are the major sites of recognition by the host immune system and these antigens are potential candidates for biological markers. The overall objective of the proposed research is to develop sensitive, specific and precise diagnostic markers for the gram-positive bacterium C. pseudotuberculosis. The specific objectives are: (1) Isolation of the major cell wall antigens (proteins, glycoproteins, glycolipids) of C. pseudotuberculosis; (2) Production of antisera against whole cells and cell wall fractions of C. pseudotuberculosis. The initial research in the development of biological markers will involve the isolation of the major cell wall proteins of C. pseudotuberculosis by mechanical disruption of the cells. The characterization of these antigens will be accomplished by the use of one and two-dimensional polyacrylamide gel electrophoresis. The immunological characterization of the surface antigens of C. pseudotuberculosis will be achieved by radioimmunoassay methodologies and Western blotting. Research in the area of health and disease will involve and depend upon a high degree of cooperation among scientists in several disciplines at Prairie View A & M University and with scientists at Texas A & M University. The proposed project will strengthen research capabilities in the biomedical sciences at PVAMU by expanding research opportunities and encouraging ethnic minorities to choose and participate in biomedical research careers.
