1887

Abstract

There is growing evidence to suggest that the resident microflora of chronic venous leg ulcers impairs cellular wound-healing responses, thereby playing an important role in maintaining the non-healing phenotype of many of these wounds. The significance of individual species of bacteria will remain unclear until it is possible to characterize fully the microflora of such lesions. The limitations and biases of culture-based microbiology are being realized and the subsequent application of molecular methods is revealing greater diversity within mixed bacterial populations than that demonstrated by culture alone. To date, this approach has been limited to a small number of systems, including the oral microflora. Here, for the first time, the comprehensive characterization of the microflora present in the tissue of a chronic venous leg ulcer is described by the comparison of 16S rDNA sequences amplified directly from the wound tissue with sequences obtained from bacteria that were isolated by culture. The molecular approach demonstrated significantly greater bacterial diversity than that revealed by culture. Furthermore, sequences were retrieved that may possibly represent novel species of bacteria. It is only by the comprehensive analysis of the wound microflora by both molecular and cultural methods that it will be possible to further our understanding of the role of bacteria in this important condition.

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2003-04-01
2024-04-16
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References

  1. Becker M. R, Paster B. J, Leys E. J, Moeschberger M. L, Kenyon S. G, Galvin J. L, Boches S. K, Dewhirst F. E, Griffen A. L. 2002; Molecular analysis of bacterial species associated with childhood caries. J Clin Microbiol 40:1001–1009
    [Google Scholar]
  2. Bowden G. H, Hardie J. M. 1971; Anaerobic organisms from the mouth. In Isolation of Anaerobes pp 177–205 Edited by Shapton D. A., Board R. G. London: Academic Press;
    [Google Scholar]
  3. Bowler P. G. 1998; The anaerobic and aerobic microbiology of wounds: a review. Wounds 10:170–178
    [Google Scholar]
  4. Bowler P. G, Davies B. J. 1999; The microbiology of acute and chronic wounds. Wounds 11:72–78
    [Google Scholar]
  5. Carroll N. M, Jaeger E. E, Choudhury S, Dunlop A. A, Matheson M. M, Adamson P, Okhravi N, Lightman S. 2000; Detection of and discrimination between gram-positive and gram-negative bacteria in intraocular samples by using nested PCR. J Clin Microbiol 38:1753–1757
    [Google Scholar]
  6. Dymock D, Weightman A. J, Scully C, Wade W. G. 1996; Molecular analysis of microflora associated with dentoalveolar abscesses. J Clin Microbiol 34:537–542
    [Google Scholar]
  7. Falanga V. 1993; Chronic wounds: pathophysiologic and experimental considerations. J Invest Dermatol 100:721–725 [CrossRef]
    [Google Scholar]
  8. Halbert A. R, Stacey M. C, Rohr J. B, Jopp-McKay A. 1992; The effect of bacterial colonization on venous ulcer healing. Australas J Dermatol 33:75–80 [CrossRef]
    [Google Scholar]
  9. Hansson C, Hoborn J, Moller A, Swanbeck G. 1995; The microbial flora in venous leg ulcers without clinical signs of infection. Repeated culture using a validated standardised microbiological technique. Acta Derm Venereol 75:24–30
    [Google Scholar]
  10. Hill K. E, Marchesi J. R, Weightman A. J. 1999; Investigation of two evolutionarily unrelated halocarboxylic acid dehalogenase gene families. J Bacteriol 181:2535–2547
    [Google Scholar]
  11. Hugenholz P, Pace N. R. 1996; Identifying microbial diversity in the natural environment: a molecular phylogenetic approach. Trends Biotechnol 14:190–197 [CrossRef]
    [Google Scholar]
  12. Keay S, Zhang C.-O, Baldwin B. R, Alexander R. B, Warren J. W. 1998; Polymerase chain reaction amplification of bacterial 16S rRNA genes from cold-cup biopsy forceps. J Urol 160:2229–2231 [CrossRef]
    [Google Scholar]
  13. Klausegger A, Hell M, Berger A, Zinober K, Baier S, Jones N, Sperl W, Kofler B. 1999; Gram type-specific broad-range PCR amplification for rapid detection of 62 pathogenic bacteria. J Clin Microbiol 37:464–466
    [Google Scholar]
  14. Kroes I, Lepp P. W, Relman D. A. 1999; Bacterial diversity within the human subgingival crevice. Proc Natl Acad Sci U S A 96:14547–14552 [CrossRef]
    [Google Scholar]
  15. Lane D. J. 1991; 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics pp 115–175 Edited by Stackebrandt E., Goodfellow M. London: Wiley;
    [Google Scholar]
  16. Marchesi J. R, Sato T, Weightman A. J, Martin T. A, Fry J. C, Hiom S. J, Dymock D, Wade W. G. 1998; Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA. Appl Environ Microbiol 64:795–799
    [Google Scholar]
  17. Murdoch D. A, Mitchelmore I. J, Tabaqchali S. 1994; The clinical importance of gram-positive anaerobic cocci isolated at St Bartholomew's Hospital. London: in 1987 J Med Microbiol 41:36–44 [CrossRef]
    [Google Scholar]
  18. Paster B. J, Boches S. K, Galvin J. L, Ericson R. E, Lau C. N, Levanos V. A, Sahasrabudhe A, Dewhirst F. E. 2001; Bacterial diversity in human subgingival plaque. J Bacteriol 183:3770–3783 [CrossRef]
    [Google Scholar]
  19. Rolph H. J, Lennon A, Riggio M. P, Saunders W. P, MacKenzie D, Coldero L, Bagg J. 2001; Molecular identification of microorganisms from endodontic infections. J Clin Microbiol 39:3282–3289 [CrossRef]
    [Google Scholar]
  20. Sambrook J, Fritsch E. F, Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  21. Suzuki M. T, Giovannoni S. J. 1996; Bias caused by template annealing in the amplification of mixtures of 16S rRNA genes by PCR. Appl Environ Microbiol 62:625–630
    [Google Scholar]
  22. Wilson M. J, Weightman A. J, Wade W. G. 1997; Applications of molecular ecology in the characterisation of uncultured microorganisms associated with human disease. Rev Med Microbiol 8:91–101 [CrossRef]
    [Google Scholar]
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