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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Nov;33(11):2872–2875. doi: 10.1128/jcm.33.11.2872-2875.1995

Comparison of Legionella pneumophila isolates by arbitrarily primed PCR and pulsed-field gel electrophoresis: analysis from seven epidemic investigations.

J M Pruckler 1, L A Mermel 1, R F Benson 1, C Giorgio 1, P K Cassiday 1, R F Breiman 1, C G Whitney 1, B S Fields 1
PMCID: PMC228598  PMID: 8576337

Abstract

Arbitrarily primed PCR (AP-PCR) and pulsed-field gel electrophoresis (PFGE) subtyping were applied to clinical and environmental isolates from seven unrelated outbreaks of Legionnaires' disease. The patterns observed with each method matched patient isolates and the epidemiologically linked source of disease for each of the seven outbreaks. PFGE allowed more discrimination among various isolates, although AP-PCR usually gave comparable results. With both methods, certain patterns appeared to predominate in the comparison of the seven outbreaks. Of five clinical isolates not associated with the outbreaks, three gave profiles distinct from those observed in the outbreaks by both methods. This suggests that there are at least two predominant subtypes of Legionella pneumophila serogroup 1 associated with outbreaks. Investigations of outbreaks of legionellosis should employ either PFGE or AP-PCR in addition to monoclonal antibody analysis.

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Selected References

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  1. Breiman R. F., Cozen W., Fields B. S., Mastro T. D., Carr S. J., Spika J. S., Mascola L. Role of air sampling in investigation of an outbreak of legionnaires' disease associated with exposure to aerosols from an evaporative condenser. J Infect Dis. 1990 Jun;161(6):1257–1261. doi: 10.1093/infdis/161.6.1257. [DOI] [PubMed] [Google Scholar]
  2. Breiman R. F., Fields B. S., Sanden G. N., Volmer L., Meier A., Spika J. S. Association of shower use with Legionnaires' disease. Possible role of amoebae. JAMA. 1990 Jun 6;263(21):2924–2926. [PubMed] [Google Scholar]
  3. Georghiou P. R., Doggett A. M., Kielhofner M. A., Stout J. E., Watson D. A., Lupski J. R., Hamill R. J. Molecular fingerprinting of Legionella species by repetitive element PCR. J Clin Microbiol. 1994 Dec;32(12):2989–2994. doi: 10.1128/jcm.32.12.2989-2994.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gomez-Lus P., Fields B. S., Benson R. F., Martin W. T., O'Connor S. P., Black C. M. Comparison of arbitrarily primed polymerase chain reaction, ribotyping, and monoclonal antibody analysis for subtyping Legionella pneumophila serogroup 1. J Clin Microbiol. 1993 Jul;31(7):1940–1942. doi: 10.1128/jcm.31.7.1940-1942.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gordillo M. E., Singh K. V., Murray B. E. Comparison of ribotyping and pulsed-field gel electrophoresis for subspecies differentiation of strains of Enterococcus faecalis. J Clin Microbiol. 1993 Jun;31(6):1570–1574. doi: 10.1128/jcm.31.6.1570-1574.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Grattard F., Pozzetto B., Berthelot P., Rayet I., Ros A., Lauras B., Gaudin O. G. Arbitrarily primed PCR, ribotyping, and plasmid pattern analysis applied to investigation of a nosocomial outbreak due to Enterobacter cloacae in a neonatal intensive care unit. J Clin Microbiol. 1994 Mar;32(3):596–602. doi: 10.1128/jcm.32.3.596-602.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Grimont F., Lefèvre M., Ageron E., Grimont P. A. rRNA gene restriction patterns of Legionella species: a molecular identification system. Res Microbiol. 1989 Nov-Dec;140(9):615–626. doi: 10.1016/0923-2508(89)90193-9. [DOI] [PubMed] [Google Scholar]
  8. Joly J. R., McKinney R. M., Tobin J. O., Bibb W. F., Watkins I. D., Ramsay D. Development of a standardized subgrouping scheme for Legionella pneumophila serogroup 1 using monoclonal antibodies. J Clin Microbiol. 1986 Apr;23(4):768–771. doi: 10.1128/jcm.23.4.768-771.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Linhardt F., Ziebuhr W., Meyer P., Witte W., Hacker J. Pulsed-field gel electrophoresis of genomic restriction fragments as a tool for the epidemiological analysis of Staphylococcus aureus and coagulase-negative staphylococci. FEMS Microbiol Lett. 1992 Aug 15;74(2-3):181–185. doi: 10.1016/0378-1097(92)90426-o. [DOI] [PubMed] [Google Scholar]
  10. Lück P. C., Helbig J. H., Günter U., Assmann M., Blau R., Koch H., Klepp M. Epidemiologic investigation by macrorestriction analysis and by using monoclonal antibodies of nosocomial pneumonia caused by Legionella pneumophila serogroup 10. J Clin Microbiol. 1994 Nov;32(11):2692–2697. doi: 10.1128/jcm.32.11.2692-2697.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mamolen M., Breiman R. F., Barbaree J. M., Gunn R. A., Stone K. M., Spika J. S., Dennis D. T., Mao S. H., Vogt R. L. Use of multiple molecular subtyping techniques to investigate a Legionnaires' disease outbreak due to identical strains at two tourist lodges. J Clin Microbiol. 1993 Oct;31(10):2584–2588. doi: 10.1128/jcm.31.10.2584-2588.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Marston B. J., Lipman H. B., Breiman R. F. Surveillance for Legionnaires' disease. Risk factors for morbidity and mortality. Arch Intern Med. 1994 Nov 14;154(21):2417–2422. [PubMed] [Google Scholar]
  13. Nolte F. S., Conlin C. A., Roisin A. J., Redmond S. R. Plasmids as epidemiological markers in nosocomial Legionnaires' disease. J Infect Dis. 1984 Feb;149(2):251–256. doi: 10.1093/infdis/149.2.251. [DOI] [PubMed] [Google Scholar]
  14. Poddar S. K. A modified method of genomic DNA preparation in agarose inserts for pulse field gel electrophoresis. Electrophoresis. 1991 Sep;12(9):674–675. doi: 10.1002/elps.1150120914. [DOI] [PubMed] [Google Scholar]
  15. Poh C. L., Yap S. C., Yeo M. Pulsed-field gel electrophoresis for differentiation of hospital isolates of Klebsiella pneumoniae. J Hosp Infect. 1993 Jun;24(2):123–128. doi: 10.1016/0195-6701(93)90074-a. [DOI] [PubMed] [Google Scholar]
  16. Salama S. M., Tabor H., Richter M., Taylor D. E. Pulsed-field gel electrophoresis for epidemiologic studies of Campylobacter hyointestinalis isolates. J Clin Microbiol. 1992 Aug;30(8):1982–1984. doi: 10.1128/jcm.30.8.1982-1984.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sanden G. N., Cassiday P. K., Barbaree J. M. Rapid immunodot technique for identifying Bordetella pertussis. J Clin Microbiol. 1993 Jan;31(1):170–172. doi: 10.1128/jcm.31.1.170-172.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schoonmaker D., Heimberger T., Birkhead G. Comparison of ribotyping and restriction enzyme analysis using pulsed-field gel electrophoresis for distinguishing Legionella pneumophila isolates obtained during a nosocomial outbreak. J Clin Microbiol. 1992 Jun;30(6):1491–1498. doi: 10.1128/jcm.30.6.1491-1498.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Struelens M. J., Maes N., Rost F., Deplano A., Jacobs F., Liesnard C., Bornstein N., Grimont F., Lauwers S., McIntyre M. P. Genotypic and phenotypic methods for the investigation of a nosocomial Legionella pneumophila outbreak and efficacy of control measures. J Infect Dis. 1992 Jul;166(1):22–30. doi: 10.1093/infdis/166.1.22. [DOI] [PubMed] [Google Scholar]
  20. Williams J. G., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. doi: 10.1093/nar/18.22.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. van Belkum A., Struelens M., Quint W. Typing of Legionella pneumophila strains by polymerase chain reaction-mediated DNA fingerprinting. J Clin Microbiol. 1993 Aug;31(8):2198–2200. doi: 10.1128/jcm.31.8.2198-2200.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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