Hostname: page-component-78c5997874-lj6df Total loading time: 0 Render date: 2024-11-14T04:36:53.517Z Has data issue: false hasContentIssue false
  • English
  • Français

Laboratory diagnosis of Mycoplasma pneumoniae infection: 1. Direct detection of antigen in respiratory exudates by enzyme immunoassay.

Published online by Cambridge University Press:  15 May 2009

T. W. Kok ,
G. Varkanis ,
B. P. Marmion ,
J. Martin  and
A. Esterman
T. W. Kok
Affiliation:
Division of Medical Virology, Institute of Medical and Veterinary Science, Adelaide and Department of Pathology, University of Adelaide.
G. Varkanis
Affiliation:
Division of Medical Virology, Institute of Medical and Veterinary Science, Adelaide and Department of Pathology, University of Adelaide.
B. P. Marmion*
Affiliation:
Division of Medical Virology, Institute of Medical and Veterinary Science, Adelaide and Department of Pathology, University of Adelaide.
J. Martin
Affiliation:
Department of Pulmonary Medicine, Adelaide Children's Hospital.
A. Esterman
Affiliation:
Epidemiology Branch, South Australian Health Commission, Adelaide.
*
*Please address correspondence and reprint requests to Professor B. P. Marmion, Medical Virology, Institute of Medical and Veterinary Science, Box 14, Rundle Mall Post Office, Adelaide, South Australia, 5000.
Rights & Permissions [Opens in a new window]

Summary

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Direct and indirect antigen capture enzyme immunoassays (Ag-EIA) have been developed for the detection of Mycoplasma pneumoniae in nasopharyngeal aspirates or sputum from respiratory infection. The sensitivity of the two Ag-EIA were similar, but the indirect method using polyclonal rabbit and guinea-pig antisera was more convenient. The Ag-EIA had a detection limit of 104–4·5 colony-forming units/ml of sample. It was specific for M. pneumoniae and gave a low level response with M. genitalium. There were no cross-reactions with 10 other species of mycoplasmas. Tests with a wide range of bacteria and chlamydia group antigen, representing agents sometimes found in the respiratory tract, were also negative. At the current level of development, the Ag-EIA detected about 90% of specimens that were also positive for culture; 43% of specimens from culture-negative-seropositive patients gave a positive result. The overall pattern of results indicated that while antigen detection is a quick and effective substitute for the slow culture method, serological examination for specific IgM antibody is also necessary to give a complete diagnostic coverage.

Type
Research Article
Information
Epidemiology & Infection , Volume 101 , Issue 3 , December 1988 , pp. 669 - 684
Copyright
Copyright © Cambridge University Press 1988

References

REFERENCES

Allen, P. Z. & Prescott, B. (1978). Immunochemical studies on a Mycoplasma pneumoniae polysaccharide fraction: cross-reactions with type 23 and 32 antipneumococeal rabbit sera. Infection & Immunity 20, 421–429.CrossRefGoogle ScholarPubMed
Avrameas, S. & Ternynck, T. (1969). The cross-linking of proteins with glutaraldehyde and its use for the preparation of immunoabsorbents. Immunochemistry 6, 53–66.CrossRefGoogle Scholar
Avrameas, S. & Ternynck, T. (1971). Peroxidase labelled antibody and Fab conjugates with enhanced intracellular penetration. Immunochemistry 8, 1175–1179.Google Scholar
Chanock, R. M., Cook, M. K., Fox, H. H., Parrot, R. H., & Huebner, R. J. (1960). Serologic evidence of infection with Eaton agent in lower respiratory illness in childhood. New England Journal of Medicine 262, 648–654.CrossRefGoogle ScholarPubMed
Chanock, R. M., Mufson, M. A., Bloom, H. H., James, W. D., Fox, H. H. & Kingston, J. R., (1961). Eaton agent pneumonia 1. Ecology of infection in military recruit population. Journal of American Medical Association 175, 213–220.Google Scholar
Clark, H. W., Fowler, R. C. & Brown, T. M. (1961). Preparation of pleuropneumonia organisms for microscopic study. Journal of Bacteriology 81, 500–502.CrossRefGoogle ScholarPubMed
Clyde, W. A. Jr. (1985). M. pneumoniae in the pediatric population. In Mycoplasmas as Agents of Human Disease, pp. 51–64. Waukegon, Illinois: Windermere Communications Inc.Google Scholar
Collier, A. M. & Clyde, W. A. Jr (1974). Appearance of Mycoplasma pneumoniae in lungs of experimentally infected hamsters and sputum from patients with natural disease. American Review of Respiratory Diseases 110, 765–773.Google ScholarPubMed
Costea, N., Yakulis, V. J. & Heller, P. (1971). The mechanism of induction of cold agglutinins by Mycoplasma pneumoniae. Journal of Immunology 106, 598–604.CrossRefGoogle Scholar
Couch, R. B., Gate, T. R. & Chanock, R. M. (1964). Infection with artificially propagated Eaton Agent (Mycoplasma pneumoniae). Journal of the American Medical Association 187, 146–151.Google ScholarPubMed
Cruickshank, R., Duguid, J. P., Marmion, B. P. & Swain, R. H. A. (1975). vol. II, The Practice of Medical Microbiology. Churchill Livingstone.Google Scholar
Dowdle, W. I. & Robinson, R. Q. (1964). An indirect haemagglutination test for the diagnosis of Mycoplasma pneumoniae infection. Proceedings of the Society of Experimental Biology & Medicine 116, 947–950.CrossRefGoogle Scholar
Eaton, M. D., Meiklejohn, G. & Van, Herick W. (1944). Studies on the etiology of primary atypical pneumonia. I. A filterable agent transmissible to cotton rats, hamsters and chick embryos. Journal of Experimental Medicine 79, 649–668.CrossRefGoogle Scholar
Foy, J. H., Kenny, G. E., Cooney, M. K. & Allan, I. D. (1979). Long term epidemiology of infections with Mycoplasma pneumoniae. Journal of Infectious Diseases 139, 681–687.CrossRefGoogle ScholarPubMed
Gosting, L. H., Cabrian, K., Sturge, J. C. & Goldstein, L. C. (1984). Identification of a species-specific antigen in Legionella pneumophila by a monoclonal antibody. Journal of Clinical Microbiology 20, 1031–1035.Google Scholar
Grady, G. F. & Gilfillan, R. F. (1979). Relation of Mycoplasma pneumoniae seroreactivity, immunosuppression and chronic disease to Legionnaires' Disease. Annals of Internal Medicine, 90, 607–610.Google Scholar
Grayston, J. T., Foy, H. M. & Kenny, G. (1969) The epidemiology of Mycoplasma infections of the human respiratory tract In The Mycoplasmatales and the l-phase of bacteria (ed. Hayflick, ), pp. 651–682. Amsterdam: North-Holland.Google Scholar
Harris, R., Marmion, B. P., Castelli, K., Hahn, C., Varkanis, G., Kok, T., Lunn, B., & Martin, J. (1988). Laboratory diagnosis of Mycoplasma pneumonia infenction. 2. Comparison of methods for the direct detection of specific antigen or nucleic acid sequences in respiratory exudates. Epidemiology & Infenction 101, 685–694CrossRefGoogle ScholarPubMed
Helbig, J. H. & Witzleb, W. (1984). Enzyme-linked immunosorbent assay (ELISA) zum Antigennachweis von Mycoplasma pneumoniae. Zeitschrift für die Gesamte Hygiene und ihre Grenzgebiete 30, 106–107.Google ScholarPubMed
Hers, J. F. Ph.. (1963). Fluorescent antibody technique in respiratory viral diseases. American Review of Respiratory Diseases 88, 316–338.Google Scholar
Hers, J. F.Ph.. & Masurel, N. (1967). Infenction with Mycoplasma pneumoniae in civilians in the Netherlands. Annals of the New York Academcy of Science 143, 447–460.Google Scholar
Hers, J. F. Ph.., Masurel, N. & Gans, J. C. (1969). Acute respiratory disease associated with pulmonary involvement in military servicemen in the Netherlands. American Review of Respiratory Diseases 100, 499–506.Google Scholar
Lind, K., Lindhardt, B. O., Schutten, H. J., Blom, J. & Christiansen, C. (1984). Serological cross reactions between Mycoplasma genitalium and Mycoplasma pneumoniae. Journal of Clinical Microbiology 20, 1036–1042.CrossRefGoogle ScholarPubMed
Madsen, R. D., Weiner, L. B., McMillan, J. A., Saeed, F. A., North, J. A. & Coates, S. R. (1988). Direct detection of Mycoplasma pneumoniae antigen in clinical specimens by a monoclonal antibody immunoblot assay. American Journal of Clinical Pathology 89, 95–99.CrossRefGoogle ScholarPubMed
Marmion, B. P., Plackett, P. & Lemcke, R. (1967). Immunochemical analysis of Mycoplasma pneumoniae. 1. Methods of extraction and reaction of fractions from Mycoplasma pneumoniae and from Mycoplasma mycoides with homologous antisera and with antisera against Streptococcus MG. Australian Journal of Experimental Biology and Medical Science 45. 163–187.CrossRefGoogle ScholarPubMed
O'Sullivan, M. T. & Marks, V. (1981). Methods for the preparation of enzyme-antibody conjugates for use in enzyme immunoassay. Methods in Enzymology 73, 147–166.CrossRefGoogle ScholarPubMed
Plackett, P., Marmion, B. P., Shaw, E. J. & Lemcke, R. M. (1969). Immunochemical analysis of Mycoplasma pneumoniae. III. Separation and chemical identification of serologically active lipids. Australian Journal of Experimental Biology and Medical Science 47, 171–195.CrossRefGoogle Scholar
Taylor-Robinson, D., Sobeslavsky, O., Jensen, K. E, Senterfit, L. B. & Chanock, R. M. (1966). Serological response to Mycoplasma pneumoniae infection. T. Evaluation of immunofluorescence, complement fixation, indirect haemagglutination and tetrazolium reduction inhibition tests for the diagnosis of infection. American Journal of Epidemiology 83. 287–298.CrossRefGoogle Scholar
Tully, J. G., Rose, D. L., Whitcomb, R. F. & Wenzel, R. P. (1979). Enhanced isolation of Mycoplasma pneumoniae from throat washings with a newly modified culture medium. Journal of Infectious Diseases 139, 478–482.Google Scholar
Tully, J. G., Whitcomb, R. F., Clark, H. F. & Williamson, D. L. (1977). Pathogenic mycoplasmas: cultivation and vertebrate pathogenicity of a new spiroplasma. Science 195. 892–894.CrossRefGoogle ScholarPubMed
Weiner, L., McMillan, J., Poe, L., Gidding, S., Lamparella, V. & Patti, A. (1985). Mycoplasma pneumoniae (Mpn) etiology of pharyngitis. Use of enzyme-immunoassay (EIA) for rapid and direct detection of infection. Abstract 1177, Pediatric Research 19, 307 A.CrossRefGoogle Scholar
Wiernik, A., Jarstraud, C. & Tunevall, G. (1978). The value of immunoelectroosmophoresis (IEOP) for etiological diagnosis of acute respiratory tract infections due to pneumococci and Mycoplasma pneumoniae. Scandinavian Journal of Infectious Diseases 10, 173–176.CrossRefGoogle ScholarPubMed