Abstract

The goal of this research project is to develop sensitive immunoassays to measure antibody (Ab) against well-defined antigens of Candida albicans and to detect such antigens in sera of infected patients at concentrations less than 40 ng/ml. An enzyme-linked immunosorbent assay (ELISA) has been devised to quantitate serum levels of Ab against Candida cell wall mannan. This ELISA is being used to measure anti-cell wall mannan Ab levels in sequential sera from patients who are at risk for developing severe Candida infections. An ELISA will be developed which can detect cell wall mannan in sera from infected patients. A major cytoplasmic antigen (mAg) of Candida has been discovered and partially characterized. Purified preparations of mAg have been obtained from cytoplasmic extracts of Candida using DEAE-cellulose and concanavalin A-sepharose chromatography. Antisera against purified mAg will be produced in rabbits and used to construct double-antibody sandwich ELISA's that can quantitate anti-mAg Ab levels and detect mAg in human sera. The usefulness of these ELISA's in the diagnosis of deep-seated Candida infections in humans will then be studied.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI015682-07
Application #
3126350
Study Section
Bacteriology and Mycology Subcommittee 1 (BM)
Project Start
1979-05-01
Project End
1987-08-31
Budget Start
1985-09-01
Budget End
1986-08-31
Support Year
7
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

Klein, B S; Jones, J M (1990) Isolation, purification, and radiolabeling of a novel 120-kD surface protein on Blastomyces dermatitidis yeasts to detect antibody in infected patients. J Clin Invest 85:152-61
Kahn, F W; Jones, J M (1988) Analysis of bronchoalveolar lavage specimens from immunocompromised patients with a protocol applicable in the microbiology laboratory. J Clin Microbiol 26:1150-5
Kahn, F W; Jones, J M (1987) Diagnosing bacterial respiratory infection by bronchoalveolar lavage. J Infect Dis 155:862-9
Olivo, P D; McManus, E J; Riggsby, W S et al. (1987) Mitochondrial DNA polymorphism in Candida albicans. J Infect Dis 156:214-5
Kahn, F W; Jones, J M (1986) Latex agglutination tests for detection of Candida antigens in sera of patients with invasive candidiasis. J Infect Dis 153:579-85
Chen, C M; Misra, T K; Silver, S et al. (1986) Nucleotide sequence of the structural genes for an anion pump. The plasmid-encoded arsenical resistance operon. J Biol Chem 261:15030-8
Ponton, J; Jones, J M (1986) Identification of two germ-tube-specific cell wall antigens of Candida albicans. Infect Immun 54:864-8
McManus, E J; Jones, J M (1986) The use of ketoconazole in the treatment of blastomycosis. Am Rev Respir Dis 133:141-3
Witte, W; Green, L; Misra, T K et al. (1986) Resistance to mercury and to cadmium in chromosomally resistant Staphylococcus aureus. Antimicrob Agents Chemother 29:663-9
Ponton, J; Jones, J M (1986) Analysis of cell wall extracts of Candida albicans by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques. Infect Immun 53:565-72

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