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. 1997 Dec;35(12):3054–3057. doi: 10.1128/jcm.35.12.3054-3057.1997

Identification of Legionella species by lipopolysaccharide antigen pattern.

D Jürgens 1, F J Fehrenbach 1
PMCID: PMC230121  PMID: 9399493

Abstract

Electrophoretic analysis of lipopolysaccharide (LPS) extracts from 430 previously serotyped Legionella isolates and 28 American Type Culture Collection (ATCC) non-Legionella pneumophila Legionella reference strains representing different Legionella species and serogroups has been performed. LPS was prepared from Legionella suspensions by sonication and proteinase K digestion. Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis, LPS bands were either stained with silver nitrate or transferred onto a nitrocellulose membrane and detected with rabbit antibodies raised against L. pneumophila serogroup 5, which was known to cross-react with L. pneumophila serogroups 1 to 14. Silver staining revealed that each of the 28 ATCC non-L. pneumophila Legionella strains possessed an individual and characteristic LPS banding pattern. The LPS profile was defined by the molecular weight of the visualized bands and/or the individual ladder-like LPS pattern. It was demonstrated by immunoblotting that non-L. pneumophila Legionella strains did not react with the serogroup 5 antiserum, thus allowing for the differentiation between L. pneumophila and non-L. pneumophila species.

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Selected References

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  1. Aucken H. M., Pitt T. L. Lipopolysaccharide profile typing as a technique for comparative typing of gram-negative bacteria. J Clin Microbiol. 1993 May;31(5):1286–1289. doi: 10.1128/jcm.31.5.1286-1289.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barthe C., Joly J. R., Ramsay D., Boissinot M., Benhamou N. Common epitope on the lipopolysaccharide of Legionella pneumophila recognized by a monoclonal antibody. J Clin Microbiol. 1988 May;26(5):1016–1023. doi: 10.1128/jcm.26.5.1016-1023.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collins M. T., Espersen F., Høiby N., Cho S. N., Friis-Møller A., Reif J. S. Cross-reactions between Legionella pneumophila (serogroup 1) and twenty-eight other bacterial species, including other members of the family Legionellaceae. Infect Immun. 1983 Mar;39(3):1441–1456. doi: 10.1128/iai.39.3.1441-1456.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Darveau R. P., Hancock R. E. Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains. J Bacteriol. 1983 Aug;155(2):831–838. doi: 10.1128/jb.155.2.831-838.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Edelstein P. H. Improved semiselective medium for isolation of Legionella pneumophila from contaminated clinical and environmental specimens. J Clin Microbiol. 1981 Sep;14(3):298–303. doi: 10.1128/jcm.14.3.298-303.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ehret W., Jacob K., Ruckdeschel G. Identification of clinical and environmental isolates of Legionella pneumophila by analysis of outer-membrane proteins, ubiquinones and fatty acids. Zentralbl Bakteriol Mikrobiol Hyg A. 1987 Aug;266(1-2):261–275. doi: 10.1016/s0176-6724(87)80040-8. [DOI] [PubMed] [Google Scholar]
  7. Ehret W., Ruckdeschel G. Species specific membrane proteins of Legionellaceae. Zentralbl Bakteriol Mikrobiol Hyg A. 1983 Jul;255(1):33–38. [PubMed] [Google Scholar]
  8. Gabay J. E., Horwitz M. A. Isolation and characterization of the cytoplasmic and outer membranes of the Legionnaires' disease bacterium (Legionella pneumophila). J Exp Med. 1985 Feb 1;161(2):409–422. doi: 10.1084/jem.161.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hindahl M. S., Iglewski B. H. Outer membrane proteins from Legionella pneumophila serogroups and other Legionella species. Infect Immun. 1986 Jan;51(1):94–101. doi: 10.1128/iai.51.1.94-101.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jürgens D., Fehrenbach F. J. Cross-reacting lipopolysaccharide antigens in Legionella pneumophila serogroups 1 to 14. Infect Immun. 1995 Jun;63(6):2180–2184. doi: 10.1128/iai.63.6.2180-2184.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kelly N. M., MacDonald M. H., Martin N., Nicas T., Hancock R. E. Comparison of the outer membrane protein and lipopolysaccharide profiles of mucoid and nonmucoid Pseudomonas aeruginosa. J Clin Microbiol. 1990 Sep;28(9):2017–2021. doi: 10.1128/jcm.28.9.2017-2021.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lambert M. A., Moss C. W. Cellular fatty acid compositions and isoprenoid quinone contents of 23 Legionella species. J Clin Microbiol. 1989 Mar;27(3):465–473. doi: 10.1128/jcm.27.3.465-473.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mills S. D., Kurjanczyk L. A., Penner J. L. Antigenicity of Helicobacter pylori lipopolysaccharides. J Clin Microbiol. 1992 Dec;30(12):3175–3180. doi: 10.1128/jcm.30.12.3175-3180.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Palva E. T., Mäkelä P. H. Lipopolysaccharide heterogeneity in Salmonella typhimurium analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Eur J Biochem. 1980;107(1):137–143. doi: 10.1111/j.1432-1033.1980.tb04634.x. [DOI] [PubMed] [Google Scholar]
  17. Petitjean F., Guillet J. G., Vray B., Strosberg A. D., Hoebeke J. Partial characterization of a Legionella pneumophila serogroup 1 immunodominant antigenic determinant recognized by a monoclonal antibody. Legionella specific antigenic determinant. Comp Immunol Microbiol Infect Dis. 1987;10(1):9–23. doi: 10.1016/0147-9571(87)90036-1. [DOI] [PubMed] [Google Scholar]
  18. Rimler R. B. Comparisons of Pasteurella multocida lipopolysaccharides by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to determine relationship between group B and E hemorrhagic septicemia strains and serologically related group A strains. J Clin Microbiol. 1990 Apr;28(4):654–659. doi: 10.1128/jcm.28.4.654-659.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tenover F. C., Edelstein P. H., Goldstein L. C., Sturge J. C., Plorde J. J. Comparison of cross-staining reactions by Pseudomonas spp. and fluorescein-labeled polyclonal and monoclonal antibodies directed against Legionella pneumophila. J Clin Microbiol. 1986 Mar;23(3):647–649. doi: 10.1128/jcm.23.3.647-649.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Thacker W. L., Plikaytis B. B., Wilkinson H. W. Identification of 22 Legionella species and 33 serogroups with the slide agglutination test. J Clin Microbiol. 1985 May;21(5):779–782. doi: 10.1128/jcm.21.5.779-782.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  22. Wilkinson H. W., Fikes B. J., Cruce D. D. Indirect immunofluorescence test for serodiagnosis of Legionnaires disease: evidence for serogroup diversity of Legionnaires disease bacterial antigens and for multiple specificity of human antibodies. J Clin Microbiol. 1979 Mar;9(3):379–383. doi: 10.1128/jcm.9.3.379-383.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wong K. H., Feeley J. C. Lipopolysaccharide of Legionella as adjuvant for intrinsic and extrinsic antigens. Proc Soc Exp Biol Med. 1984 Dec;177(3):475–481. doi: 10.3181/00379727-177-41975. [DOI] [PubMed] [Google Scholar]

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