Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1992 Sep;30(9):2408–2414. doi: 10.1128/jcm.30.9.2408-2414.1992

Restriction endonuclease analysis and ribotyping differentiate genital and nongenital strains of Bacteroides ureolyticus.

N Akhtar 1, A Eley 1
PMCID: PMC265514  PMID: 1401007

Abstract

Thirty-three clinical isolates from male nongonococcal urethritis and 28 isolates from soft tissue infections and ulcers were identified as Bacteroides ureolyticus by conventional bacteriological tests and were compared with five reference strains of the species. Whole-cell proteins from these clinical isolates and the reference strains were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The majority of the strains from the two sources could be divided into five different groups, named phenons I to V; phenons I to IV have been described previously by others, while phenon V has been described recently by us. Digestion of chromosomal DNA from 16 of the clinical isolates (including strains representative of each of the five SDS-PAGE phenons) and the five reference strains was attempted with restriction endonucleases EcoRI, PstI, SmaI, and HindIII. After electrophoresis in agarose gels, good digestion was observed with HindIII only, and 12 different banding patterns (restriction endonuclease analysis [REA] profiles) were obtained for the 19 strains digested; one nongonococcal urethritis isolate and one reference strain did not show any digestion. From the agarose gels, HindIII-digested fragments of DNA were transferred to nylon membranes by use of vacuum blotting and subjected to hybridization with 32P-labelled 16S-23S rRNA from Escherichia coli. The resultant pattern of bands (ribotypes), which depends on the restriction fragment length polymorphisms in the rRNA genes, was used as a measure of genomic variation within the species. In total, 13 different ribotypes were obtained for the 19 strains. For some strains, good correlation was achieved among the SDS-PAGE phenons, REA profiles, and ribotypes. However, for others, REA analysis and ribotyping were able to discriminate between strains which shared the same SDS-PAGE phenon. Interestingly, these two techniques of DNA characterization were able to differentiate between isolates from the genital tract and those associated with soft tissue infections and ulcers.

Full text

PDF
2408

Images in this article

2410Image
on p.2410
2410Image
on p.2410

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akhtar N., Eley A., Kinghorn G. R. Simple transport medium for Bacteroides ureolyticus. Eur J Clin Microbiol Infect Dis. 1989 Apr;8(4):321–322. doi: 10.1007/BF01963466. [DOI] [PubMed] [Google Scholar]
  2. Allardet-Servent A., Bouziges N., Carles-Nurit M. J., Bourg G., Gouby A., Ramuz M. Use of low-frequency-cleavage restriction endonucleases for DNA analysis in epidemiological investigations of nosocomial bacterial infections. J Clin Microbiol. 1989 Sep;27(9):2057–2061. doi: 10.1128/jcm.27.9.2057-2061.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett K. W., Eley A., Woolley P. D., Duerden B. I. Isolation of Bacteroides ureolyticus from the genital tract of men with and without non-gonococcal urethritis. Eur J Clin Microbiol Infect Dis. 1990 Nov;9(11):825–826. doi: 10.1007/BF01967383. [DOI] [PubMed] [Google Scholar]
  4. Bennett K. W., Eley A., Woolley P. D. Isolation of Bacteroides ureolyticus from the female genital tract. Eur J Clin Microbiol Infect Dis. 1991 Jul;10(7):593–594. doi: 10.1007/BF01967281. [DOI] [PubMed] [Google Scholar]
  5. Chen C. K., Sunday G. J., Zambon J. J., Wilson M. E. Restriction endonuclease analysis of Eikenella corrodens. J Clin Microbiol. 1990 Jun;28(6):1265–1270. doi: 10.1128/jcm.28.6.1265-1270.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Collins D. M., Gabric D. M., de Lisle G. W. Identification of two groups of Mycobacterium paratuberculosis strains by restriction endonuclease analysis and DNA hybridization. J Clin Microbiol. 1990 Jul;28(7):1591–1596. doi: 10.1128/jcm.28.7.1591-1596.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Duerden B. I., Eley A., Goodwin L., Magee J. T., Hindmarch J. M., Bennett K. W. A comparison of Bacteroides ureolyticus isolates from different clinical sources. J Med Microbiol. 1989 May;29(1):63–73. doi: 10.1099/00222615-29-1-63. [DOI] [PubMed] [Google Scholar]
  8. Duerden B., Bennet K. W., Faulkner J. Isolation of Bacteroides ureolyticus (B corrodens) from clinical infections. J Clin Pathol. 1982 Mar;35(3):309–312. doi: 10.1136/jcp.35.3.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eley A., Akhtar N., Bennett K. W. In-vitro activity of ten antibiotics against clinical isolates of Bacteroides ureolyticus. J Antimicrob Chemother. 1990 Apr;25(4):703–704. doi: 10.1093/jac/25.4.703. [DOI] [PubMed] [Google Scholar]
  10. Eley A., Clarry T., Bennett K. W. Selective and differential medium for isolation of Bacteroides ureolyticus from clinical specimens. Eur J Clin Microbiol Infect Dis. 1989 Jan;8(1):83–85. doi: 10.1007/BF01964126. [DOI] [PubMed] [Google Scholar]
  11. Fontaine E. A., Borriello S. P., Taylor-Robinson D., Davies H. A. Characteristics of a gram-negative anaerobe isolated from men with non-gonococcal urethritis. J Med Microbiol. 1984 Apr;17(2):129–140. doi: 10.1099/00222615-17-2-129. [DOI] [PubMed] [Google Scholar]
  12. Fontaine E. A., Bryant T. N., Taylor-Robinson D., Borriello S. P., Davies H. A. A numerical taxonomic study of anaerobic gram-negative bacilli classified as Bacteroides ureolyticus isolated from patients with non-gonococcal urethritis. J Gen Microbiol. 1986 Nov;132(11):3137–3146. doi: 10.1099/00221287-132-11-3137. [DOI] [PubMed] [Google Scholar]
  13. Fontaine E. A., Clark J. B., Abeck D., Taylor-Robinson D. The effect of a toxin from Bacteroides ureolyticus on the mucosal epithelium of human and bovine oviducts. Br J Exp Pathol. 1988 Oct;69(5):631–638. [PMC free article] [PubMed] [Google Scholar]
  14. Fontaine E. A., Taylor-Robinson D., Hanna N. F., Coufalik E. D. Anaerobes in men with urethritis. Br J Vener Dis. 1982 Oct;58(5):321–326. doi: 10.1136/sti.58.5.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Hawkins D. A., Fontaine E. A., Thomas B. J., Boustouller Y. L., Taylor-Robinson D. The enigma of non-gonococcal urethritis: role for Bacteroides ureolyticus. Genitourin Med. 1988 Feb;64(1):10–13. doi: 10.1136/sti.64.1.10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Irino K., Grimont F., Casin I., Grimont P. A. rRNA gene restriction patterns of Haemophilus influenzae biogroup aegyptius strains associated with Brazilian purpuric fever. J Clin Microbiol. 1988 Aug;26(8):1535–1538. doi: 10.1128/jcm.26.8.1535-1538.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kersters K., De Ley J. Identification and grouping of bacteria by numerical analysis of their electrophoretic protein patterns. J Gen Microbiol. 1975 Apr;87(2):333–342. doi: 10.1099/00221287-87-2-333. [DOI] [PubMed] [Google Scholar]
  19. Kirby B. D., George W. L., Sutter V. L., Citron D. M., Finegold S. M. Gram-negative anaerobic bacilli: their role in infection and patterns of susceptibility to antimicrobial agents. I. Little-known Bacteroides species. Rev Infect Dis. 1980 Nov-Dec;2(6):914–951. doi: 10.1093/clinids/2.6.914. [DOI] [PubMed] [Google Scholar]
  20. Kuijper E. J., van Alphen L., Leenders E., Zanen H. C. Typing of Aeromonas strains by DNA restriction endonuclease analysis and polyacrylamide gel electrophoresis of cell envelopes. J Clin Microbiol. 1989 Jun;27(6):1280–1285. doi: 10.1128/jcm.27.6.1280-1285.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mogollon J. D., Pijoan C., Murtaugh M. P., Kaplan E. L., Collins J. E., Cleary P. P. Characterization of prototype and clinically defined strains of Streptococcus suis by genomic fingerprinting. J Clin Microbiol. 1990 Nov;28(11):2462–2466. doi: 10.1128/jcm.28.11.2462-2466.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Owen R. J., Borman P. A rapid biochemical method for purifying high molecular weight bacterial chromosomal DNA for restriction enzyme analysis. Nucleic Acids Res. 1987 Apr 24;15(8):3631–3631. doi: 10.1093/nar/15.8.3631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Owen R. J. Chromosomal DNA fingerprinting--a new method of species and strain identification applicable to microbial pathogens. J Med Microbiol. 1989 Oct;30(2):89–99. doi: 10.1099/00222615-30-2-89. [DOI] [PubMed] [Google Scholar]
  24. Stull T. L., LiPuma J. J., Edlind T. D. A broad-spectrum probe for molecular epidemiology of bacteria: ribosomal RNA. J Infect Dis. 1988 Feb;157(2):280–286. doi: 10.1093/infdis/157.2.280. [DOI] [PubMed] [Google Scholar]
  25. Taylor A. J., Costas M., Owen R. J. Numerical analysis of electrophoretic protein patterns of Bacteroides ureolyticus clinical isolates. J Clin Microbiol. 1987 Apr;25(4):660–666. doi: 10.1128/jcm.25.4.660-666.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Taylor A. J., Dawson C. A., Owen R. J. The identification of Bacteroides ureolyticus from patients with non-gonococcal urethritis by conventional biochemical tests and by DNA and protein analyses. J Med Microbiol. 1986 Mar;21(2):109–116. doi: 10.1099/00222615-21-2-109. [DOI] [PubMed] [Google Scholar]
  27. Woese C. R. Bacterial evolution. Microbiol Rev. 1987 Jun;51(2):221–271. doi: 10.1128/mr.51.2.221-271.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES