Abstract
Typing of Legionella pneumophila remains important in the investigation of outbreaks of Legionnaires' disease and in the control of organisms contaminating hospital water. We found that the discriminatory power of a nonradioactive ribotyping method could be improved by combining results obtained with four restriction enzymes (HindIII, NciI, ClaI, and PstI). Fifty-eight clinical and environmental L. pneumophila strains including geographically unrelated as well as epidemiologically connected isolates were investigated. Epidemiologically related strains had the same ribotypes independent of the combinations of enzymes used. Some strains belonging to the same serogroup were assigned to different ribotypes, and some ribotypes contained members of different serogroups, indicating, as others have found, that serogroup and genotype are not always related. The discriminatory power of the method was estimated by calculating an index of discrimination (ID) for individual enzymes and combinations thereof. The combined result with all four enzymes was highly discriminatory (ID = 0.97), but results for three enzymes also yielded ID values acceptable for epidemiological purposes. In addition, the testing of 27 type strains and 6 clinical isolates representing Legionella species other than L. pneumophila indicated that ribotyping might be of value for species identification within this genus, as previously suggested.
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- Blanc D. S., Siegrist H. H., Sahli R., Francioli P. Ribotyping of Pseudomonas aeruginosa: discriminatory power and usefulness as a tool for epidemiological studies. J Clin Microbiol. 1993 Jan;31(1):71–77. doi: 10.1128/jcm.31.1.71-77.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brenner D. J., Steigerwalt A. G., Epple P., Bibb W. F., McKinney R. M., Starnes R. W., Colville J. M., Selander R. K., Edelstein P. H., Moss C. W. Legionella pneumophila serogroup Lansing 3 isolated from a patient with fatal pneumonia, and descriptions of L. pneumophila subsp. pneumophila subsp. nov., L. pneumophila subsp. fraseri subsp. nov., and L. pneumophila subsp. pascullei subsp. nov. J Clin Microbiol. 1988 Sep;26(9):1695–1703. doi: 10.1128/jcm.26.9.1695-1703.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Colding H., Bangsborg J., Fiehn N. E., Bennekov T., Bruun B. Ribotyping for differentiating Flavobacterium meningosepticum isolates from clinical and environmental sources. J Clin Microbiol. 1994 Feb;32(2):501–505. doi: 10.1128/jcm.32.2.501-505.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cox N., Johnston J., Szarka Z., Wright D. J., Archard L. C. Characterization of an rRNA gene-specific cDNA probe: applications in bacterial identification. J Gen Microbiol. 1990 Aug;136(8):1639–1643. doi: 10.1099/00221287-136-8-1639. [DOI] [PubMed] [Google Scholar]
- Dennis P. J., Brenner D. J., Thacker W. L., Wait R., Vesey G., Steigerwalt A. G., Benson R. F. Five new Legionella species isolated from water. Int J Syst Bacteriol. 1993 Apr;43(2):329–337. doi: 10.1099/00207713-43-2-329. [DOI] [PubMed] [Google Scholar]
- Edelstein P. H., Beer K. B., DeBoynton E. D. Influence of growth temperature on virulence of Legionella pneumophila. Infect Immun. 1987 Nov;55(11):2701–2705. doi: 10.1128/iai.55.11.2701-2705.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Gerner-Smidt P. Ribotyping of the Acinetobacter calcoaceticus-Acinetobacter baumannii complex. J Clin Microbiol. 1992 Oct;30(10):2680–2685. doi: 10.1128/jcm.30.10.2680-2685.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Grimont F., Grimont P. A. Ribosomal ribonucleic acid gene restriction patterns as potential taxonomic tools. Ann Inst Pasteur Microbiol. 1986 Sep-Oct;137B(2):165–175. doi: 10.1016/s0769-2609(86)80105-3. [DOI] [PubMed] [Google Scholar]
- 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]
- Harrison T. G., Saunders N. A., Haththotuwa A., Doshi N., Taylor A. G. Further evidence that genotypically closely related strains of Legionella pneumophila can express different serogroup specific antigens. J Med Microbiol. 1992 Sep;37(3):155–161. doi: 10.1099/00222615-37-3-155. [DOI] [PubMed] [Google Scholar]
- Harrison T. G., Saunders N. A., Haththotuwa A., Doshi N., Taylor A. G. Typing of Legionella pneumophila serogroups 2-14 strains by analysis of restriction fragment length polymorphisms. Lett Appl Microbiol. 1990 Oct;11(4):189–192. doi: 10.1111/j.1472-765x.1990.tb00157.x. [DOI] [PubMed] [Google Scholar]
- Harrison T. G., Saunders N. A., Haththotuwa A., Hallas G., Birtles R. J., Taylor A. G. Phenotypic variation amongst genotypically homogeneous Legionella pneumophila serogroup 1 isolates: implications for the investigation of outbreaks of Legionnaires' disease. Epidemiol Infect. 1990 Apr;104(2):171–180. doi: 10.1017/s0950268800059331. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hunter P. R., Gaston M. A. Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity. J Clin Microbiol. 1988 Nov;26(11):2465–2466. doi: 10.1128/jcm.26.11.2465-2466.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hunter P. R. Reproducibility and indices of discriminatory power of microbial typing methods. J Clin Microbiol. 1990 Sep;28(9):1903–1905. doi: 10.1128/jcm.28.9.1903-1905.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnston J. M., Latham R. H., Meier F. A., Green J. A., Boshard R., Mooney B. R., Edelstein P. H. Nosocomial outbreak of Legionnaires' disease: molecular epidemiology and disease control measures. Infect Control. 1987 Feb;8(2):53–58. doi: 10.1017/s0195941700067072. [DOI] [PubMed] [Google Scholar]
- 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]
- Lanser J., Adams M., Doyle R., Hewitt P., Sangster N. Genetic characterization of Legionella pneumophila serogroup 1 associated with respiratory disease in Australia. Appl Environ Microbiol. 1992 Feb;58(2):706–708. doi: 10.1128/aem.58.2.706-708.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lück P. C., Bender L., Ott M., Helbig J. H., Hacker J. Analysis of Legionella pneumophila serogroup 6 strains isolated from a hospital warm water supply over a three-year period by using genomic long-range mapping techniques and monoclonal antibodies. Appl Environ Microbiol. 1991 Nov;57(11):3226–3231. doi: 10.1128/aem.57.11.3226-3231.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marra A., Shuman H. A. Genetics of Legionella pneumophila virulence. Annu Rev Genet. 1992;26:51–69. doi: 10.1146/annurev.ge.26.120192.000411. [DOI] [PubMed] [Google Scholar]
- Mauchline W. S., Keevil C. W. Development of the BIOLOG substrate utilization system for identification of Legionella spp. Appl Environ Microbiol. 1991 Nov;57(11):3345–3349. doi: 10.1128/aem.57.11.3345-3349.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saunders N. A., Harrison T. G., Haththotuwa A., Kachwalla N., Taylor A. G. A method for typing strains of Legionella pneumophila serogroup 1 by analysis of restriction fragment length polymorphisms. J Med Microbiol. 1990 Jan;31(1):45–55. doi: 10.1099/00222615-31-1-45. [DOI] [PubMed] [Google Scholar]
- Saunders N. A., Harrison T. G., Haththotuwa A., Taylor A. G. A comparison of probes for restriction fragment length polymorphism (RFLP) typing of Legionella pneumophila serogroup 1 strains. J Med Microbiol. 1991 Sep;35(3):152–158. doi: 10.1099/00222615-35-3-152. [DOI] [PubMed] [Google Scholar]
- 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]
- Selander R. K., McKinney R. M., Whittam T. S., Bibb W. F., Brenner D. J., Nolte F. S., Pattison P. E. Genetic structure of populations of Legionella pneumophila. J Bacteriol. 1985 Sep;163(3):1021–1037. doi: 10.1128/jb.163.3.1021-1037.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stout J. E., Joly J., Para M., Plouffe J., Ciesielski C., Blaser M. J., Yu V. L. Comparison of molecular methods for subtyping patients and epidemiologically linked environmental isolates of Legionella pneumophila. J Infect Dis. 1988 Mar;157(3):486–495. doi: 10.1093/infdis/157.3.486. [DOI] [PubMed] [Google Scholar]
- 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]
- Verma U. K., Brenner D. J., Thacker W. L., Benson R. F., Vesey G., Kurtz J. B., Dennis P. J., Steigerwalt A. G., Robinson J. S., Moss C. W. Legionella shakespearei sp. nov., isolated from cooling tower water. Int J Syst Bacteriol. 1992 Jul;42(3):404–407. doi: 10.1099/00207713-42-3-404. [DOI] [PubMed] [Google Scholar]
- Vesey G., Dennis P. J., Lee J. V., West A. A. Further development of simple tests to differentiate the legionellas. J Appl Bacteriol. 1988 Oct;65(4):339–345. doi: 10.1111/j.1365-2672.1988.tb01900.x. [DOI] [PubMed] [Google Scholar]
- Vickers R. M., Stout J. E., Tompkins L. S., Troup N. J., Yu V. L. Cefamandole-susceptible strains of Legionella pneumophila serogroup 1: implications for diagnosis and utility as an epidemiological marker. J Clin Microbiol. 1992 Mar;30(3):537–539. doi: 10.1128/jcm.30.3.537-539.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilkinson I. J., Sangster N., Ratcliff R. M., Mugg P. A., Davos D. E., Lanser J. A. Problems associated with identification of Legionella species from the environment and isolation of six possible new species. Appl Environ Microbiol. 1990 Mar;56(3):796–802. doi: 10.1128/aem.56.3.796-802.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]