Abstract
An enzyme-linked immunosorbent assay (ELISA) with goat anti-rabbit IgG conjugated to peroxidase was used to test for the two antigen types ofPseudomonas aeruginosa: b (homogeneous) and a (heterogeneous) which contains the common subantigen (a0) and combinations of subtypes (a, a2 a3 a4). Preparations of b-type flagellar antigen could be distinguished from a-type by using b-adsorbed antisera titers as reciprocals of endpoint dilutions exceeding one million. Extracts from nonflagellated bacteria or purified lipopolysaccharides from the same strain were used as controls, which showed only background activity. Unknown flagellar antigen was determined using both isolated antigen preparations and formalin-killed bacterial cells. The ELISA procedure proved much more sensitive than the slide agglutination procedure: whereas nine of 18 strains tested did not react in the slide agglutination procedure all 18 strains were definitively typed as a or b strains with the ELISA. The ELISA also revealed the presence of a dominant, cross-reacting epitope (a0) in the heterogeneous a-type flagella using either isolated antigen or intact cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Cryz, S., Jr.: Pseudomonas aeruginosa infections. In: Germanier, R. (ed.): Bacterial vaccines. Wiley Medical, New York, p. 317–351.
Montie, T. C., Craven, R., Holder, I. A. Flagellar preparations fromPseudomonas aeruginosa: isolation and characterization. Infection and Immunity 1982, 35: 281–288.
Holder, I. A., Wheeler, R., Montie, T. C. Flagellar preparations fromPseudomonas aeruginosa: animal protection studies. Infection and Immunity 1982, 35: 276–280.
Montie, T. C., Doyle-Huntzinger, D., Craven, R., Holder, I. A. Loss of virulence associated with absence of a flagellum in an isogenic mutant ofPseudomonas aeruginosa in the burned mouse model. Infection and Immunity 1982, 38: 1296–1298.
Montie, T. C., Drake, D., Sellin, H., Slater, O., Edmonds, S. Motility virulence, and protection with a flagella vaccine againstPseudomonas aeruginosa infection. Antibiotics and Chemotherapy 1987, 39: 233–248.
Ansorg, R. Flagellaspezifisches H-Antigenschema vonPseudomonas aeruginosa. Zentralblatt für Bakteriologie, Mikrobiologie und Hygiene (A) 1978, 242: 228–238.
Lanyi, B. Serological properties ofPseudomonas aeruginosa. II. Type-specific thermolabile (flagellar) antigens. Acta Microbiologica Academiae Scientiarum Hungaricae 1970, 17: 35–48.
Penketh, A., Pitt, T., Roberts, D., Hodson, M. E., Batten, J. C. The relationship of phenotype changes inPseudomonas aeruginosa to the clinical condition of patients with cystic fibrosis. American Review of Respiratory Diseases 1983, 127: 605–608.
Allison, J. S., Dawson, M., Drake, D., Montie, T. C. Electrophoretic separation and molecular weight characterization ofPseudomonas aeruginosa H-antigen flagellins. Infection and Immunity 1985, 49: 770–774.
Pitt, T. L., Todd, H. C., Mackintosh, C. A., Im, S. W. K. Evaluation of three serological tests for detection of antibody toPseudomonas aeruginosa in human sera. European Journal of Clinical Microbiology 1985, 4: 190–196.
Jagger, R. S., Robinson, D. L., Franz, M. F., Warren, R. L. Detection by enzyme-linked immunosorbent assays of antibody specificPseudomonas proteases and exotoxin A in sera from cystic fibrosis patients. Journal of Clinical Microbiology 1982, 15: 1054–1058.
Bryan, L. E., Kureishi, A., Rabin, H. R. Detection of antibodies toPseudomonas aeruginosa alginate extracellular polysaccharide in animals and cystic fibrosis patients by enzyme-linked immunosorbent assay. Journal of Clinical Microbiology 1983, 18: 276–282.
Sordelli, D. O., Rojas, R. A., Cerquetti, M. C., Hooker, A. M., Degnan, P. J., Bellanti, J. A. Enzyme-linked immunosorbent assay antibody responses to a temperature-sensitive mutant ofPseudomonas aeruginosa. Infection and Immunity 1985, 50: 324–327.
Hancock, R. E. W., Wieczorek, A. A., Murtharia, L. M., Poole, K. Monoclonal antibodies againstPseudomonas aeruginosa outer membrane antigens: isolation and characterization. Infection and Immunity 1982, 37: 166–171.
Meadow, P. M., Rowe, P. S. N., Wells, P. L. Characterization of polyagglutinating and surface antigens inPseudomonas aeruginosa. Journal of General Microbiology 1984, 130: 631–644.
Ansorg, R. A., Krocke, M. E., Spies, A. F., Kraus, C. J. Differentiation of the major flagellar antigen ofPseudomonas aeruginosa by the slide coagglutination assay. Journal of Clinical Microbiology 1984, 20: 84–88.
Pitt, T. L. A comparison of flagellar typing and phage typing as means of subdividing the O group ofPseudomonas aeruginosa. Journal of Medical Microbiology 1981, 14: 261–270.
Bettelhelm, K. A., Maskill, W. J. Investigation by enzyme-linked immunosorbent assay ofSalmonella H antigen-antibody reactions. Journal of Clinical Microbiology 1985, 21: 772–774.
Montie, T. C., Stover, G. B. Flagellar preparation fromPseudomonas species: isolation and characterization by molecular weight. Journal of Clinical Microbiology 1983, 18: 452–456.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Montie, T.C., Anderson, T.R. Enzyme-linked immunosorbent assay for detection ofPseudomonas aeruginosa H (Flagellar) antigen. Eur. J. Clin. Microbiol. Infect. Dis. 7, 256–260 (1988). https://doi.org/10.1007/BF01963097
Issue Date:
DOI: https://doi.org/10.1007/BF01963097