1887

Abstract

Members of the complex are important bacterial pathogens in cystic fibrosis (CF) patients. The complex currently consists of nine genetic subgroups (genomovars) of different epidemiological relevance and possibly of different pathogenic potential in humans. In this study, a new approach was developed for the rapid identification of genomovar I, (genomovar II), (lineage III-A and III-B), (genomovar IV) and (genomovar V), which cause the large majority of infections in CF patients. The method was based on the detection of differences in the gene sequence by using rapid-cycle PCR and genomovar-specific fluorescence resonance energy transfer (FRET) probes. The genomovar status of all 39 complex strains tested (genomovars I–V) was identified by melting-curve analysis. Each FRET probe produced a specific fluorescence signal only with the respective genomovar, and not with other complex strains and spp. The identification system was easy to handle and revealed complex genomovar I–V status from culture isolates within about 1 h.

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2006-06-01
2019-11-19
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References

  1. Agodi, A., Mahenthiralingam, E., Barchitta, M., Giannino, V., Sciacca, A. & Stefani, S. ( 2001; ). Burkholderia cepacia complex infection in Italian patients with cystic fibrosis: prevalence, epidemiology, and genomovar status. J Clin Microbiol 39, 2891–2896.[CrossRef]
    [Google Scholar]
  2. Agodi, A., Barchitta, M., Giannino, V. & 8 other authors ( 2002; ). Burkholderia cepacia complex in cystic fibrosis and non-cystic fibrosis patients: identification of a cluster of epidemic lineages. J Hosp Infect 50, 188–195.[CrossRef]
    [Google Scholar]
  3. Aris, R. M., Routh, J. C., LiPuma, J. J., Heath, D. G. & Gilligan, P. H. ( 2001; ). Lung transplantation for cystic fibrosis patients with Burkholderia cepacia complex. Survival linked to genomovar type. Am J Respir Crit Care Med 164, 2102–2106.[CrossRef]
    [Google Scholar]
  4. Balandreau, J., Viallard, V., Cournoyer, B., Coenye, T., Laevens, S. & Vandamme, P. ( 2001; ). Burkholderia cepacia genomovar III is a common plant-associated bacterium. Appl Environ Microbiol 67, 982–985.[CrossRef]
    [Google Scholar]
  5. Bauernfeind, A., Schneider, I., Jungwirth, R. & Roller, C. ( 1999; ). Discrimination of Burkholderia multivorans and Burkholderia vietnamiensis from Burkholderia cepacia genomovars I, III, and IV by PCR. J Clin Microbiol 37, 1335–1339.
    [Google Scholar]
  6. Biddick, R., Spilker, T., Martin, A. & LiPuma, J. J. ( 2003; ). Evidence of transmission of Burkholderia cepacia, Burkholderia multivorans and Burkholderia dolosa among persons with cystic fibrosis. FEMS Microbiol Lett 228, 57–62.[CrossRef]
    [Google Scholar]
  7. Chaparro, C., Maurer, J., Gutierrez, C. & 8 other authors ( 2001; ). Infection with Burkholderia cepacia in cystic fibrosis: outcome following lung transplantation. Am J Respir Crit Care Med 163, 43–48.[CrossRef]
    [Google Scholar]
  8. Cockerill, F. R. & Smith, T. F. ( 2002; ). Rapid-cycle real-time PCR: a revolution for clinical microbiology. ASM News 68, 77–83.
    [Google Scholar]
  9. Coenye, T., Schouls, L. M., Govan, J. R., Kersters, K. & Vandamme, P. ( 1999; ). Identification of Burkholderia species and genomovars from cystic fibrosis patients by AFLP fingerprinting. Int J Syst Bacteriol 49, 1657–1666.[CrossRef]
    [Google Scholar]
  10. Coenye, T., LiPuma, J. J., Henry, D., Hoste, B., Vandemeulebroecke, K., Gillis, M., Speert, D. P. & Vandamme, P. ( 2001a; ). Burkholderia cepacia genomovar VI, a new member of the Burkholderia cepacia complex isolated from cystic fibrosis patients. Int J Syst Evol Microbiol 51, 271–279.
    [Google Scholar]
  11. Coenye, T., Mahenthiralingam, E., Henry, D., LiPuma, J. J., Laevens, S., Gillis, M., Speert, D. P. & Vandamme, P. ( 2001b; ). Burkholderia ambifaria sp. nov., a novel member of the Burkholderia cepacia complex including biocontrol and cystic fibrosis-related isolates. Int J Syst Evol Microbiol 51, 1481–1490.
    [Google Scholar]
  12. Coenye, T., Vandamme, P., Govan, J. R. & LiPuma, J. J. ( 2001c; ). Taxonomy and identification of the Burkholderia cepacia complex. J Clin Microbiol 39, 3427–3436.[CrossRef]
    [Google Scholar]
  13. Drevinek, P., Hrbackova, H., Cinek, O., Bartosova, J., Nyc, O., Nemec, A. & Pohunek, P. ( 2002; ). Direct PCR detection of Burkholderia cepacia complex and identification of its genomovars by using sputum as source of DNA. J Clin Microbiol 40, 3485–3488.[CrossRef]
    [Google Scholar]
  14. Gillis, M., Van, T. V., Bardin, R. & 7 other authors ( 1995; ). Polyphasic taxonomy in the genus Burkholderia leading to an emended description of the genus and proposition of Burkholderia vietnamiensis sp. nov. for N2-fixing isolates from rice in Vietnam. Int J Syst Bacteriol 45, 274–289.[CrossRef]
    [Google Scholar]
  15. Govan, J. R., Brown, P. H., Maddison, J., Doherty, C. J., Nelson, J. W., Dodd, M., Greening, A. P. & Webb, A. K. ( 1993; ). Evidence for transmission of Pseudomonas cepacia by social contact in cystic fibrosis. Lancet 342, 15–19.[CrossRef]
    [Google Scholar]
  16. Henry, D. A., Campbell, M. E., LiPuma, J. J. & Speert, D. P. ( 1997; ). Identification of Burkholderia cepacia isolates from patients with cystic fibrosis and use of a simple new selective medium. J Clin Microbiol 35, 614–619.
    [Google Scholar]
  17. Henry, D. A., Mahenthiralingam, E., Vandamme, P., Coenye, T. & Speert, D. P. ( 2001; ). Phenotypic methods for determining genomovar status of the Burkholderia cepacia complex. J Clin Microbiol 39, 1073–1078.[CrossRef]
    [Google Scholar]
  18. Isles, A., Maclusky, I., Corey, M., Gold, R., Prober, C., Fleming, P. & Levison, H. ( 1984; ). Pseudomonas cepacia infection in cystic fibrosis: an emerging problem. J Pediatr 104, 206–210.[CrossRef]
    [Google Scholar]
  19. LiPuma, J. J., Dulaney, B. J., McMenamin, J. D., Whitby, P. W., Stull, T. L., Coenye, T. & Vandamme, P. ( 1999; ). Development of rRNA-based PCR assays for identification of Burkholderia cepacia complex isolates recovered from cystic fibrosis patients. J Clin Microbiol 37, 3167–3170.
    [Google Scholar]
  20. LiPuma, J. J., Spilker, T., Gill, L. H., Campbell, P. W., III, Liu, L. & Mahenthiralingam, E. ( 2001; ). Disproportionate distribution of Burkholderia cepacia complex species and transmissibility markers in cystic fibrosis. Am J Respir Crit Care Med 164, 92–96.[CrossRef]
    [Google Scholar]
  21. LiPuma, J. J., Spilker, T., Coenye, T. & Gonzalez, C. F. ( 2002; ). An epidemic Burkholderia cepacia complex strain identified in soil. Lancet 359, 2002–2003.[CrossRef]
    [Google Scholar]
  22. Mahenthiralingam, E., Bischof, J., Byrne, S. K., Radomski, C., Davies, J. E., Av-Gay, Y. & Vandamme, P. ( 2000; ). DNA-based diagnostic approaches for identification of Burkholderia cepacia complex, Burkholderia vietnamiensis, Burkholderia multivorans, Burkholderia stabilis, and Burkholderia cepacia genomovars I and III. J Clin Microbiol 38, 3165–3173.
    [Google Scholar]
  23. McDowell, A., Mahenthiralingam, E., Moore, J. E. & 8 other authors ( 2001; ). PCR-based detection and identification of Burkholderia cepacia complex pathogens in sputum from cystic fibrosis patients. J Clin Microbiol 39, 4247–4255.[CrossRef]
    [Google Scholar]
  24. Moore, J. E., Xu, J., Millar, B. C., Crowe, M. & Elborn, J. S. ( 2002; ). Improved molecular detection of Burkholderia cepacia genomovar III and Burkholderia multivorans directly from sputum of patients with cystic fibrosis. J Microbiol Methods 49, 183–191.[CrossRef]
    [Google Scholar]
  25. Muhdi, K., Edenborough, F. P., Gumery, L., O'Hickey, S., Smith, E. G., Smith, D. L. & Stableforth, D. E. ( 1996; ). Outcome for patients colonised with Burkholderia cepacia in a Birmingham adult cystic fibrosis clinic and the end of an epidemic. Thorax 51, 374–377.[CrossRef]
    [Google Scholar]
  26. Segonds, C., Heulin, T., Marty, N. & Chabanon, G. ( 1999; ). Differentiation of Burkholderia species by PCR-restriction fragment length polymorphism analysis of the 16S rRNA gene and application to cystic fibrosis isolates. J Clin Microbiol 37, 2201–2208.
    [Google Scholar]
  27. Snell, G. I., de Hoyos, A., Krajden, M., Winton, T. & Maurer, J. R. ( 1993; ). Pseudomonas cepacia in lung transplant recipients with cystic fibrosis. Chest 103, 466–471.[CrossRef]
    [Google Scholar]
  28. Speert, D. P., Henry, D., Vandamme, P., Corey, M. & Mahenthiralingam, E. ( 2002; ). Epidemiology of Burkholderia cepacia complex in patients with cystic fibrosis, Canada. Emerg Infect Dis 8, 181–187.[CrossRef]
    [Google Scholar]
  29. Thomassen, M. J., Demko, C. A., Klinger, J. D. & Stern, R. C. ( 1985; ). Pseudomonas cepacia colonization among patients with cystic fibrosis. A new opportunist. Am Rev Respir Dis 131, 791–796.
    [Google Scholar]
  30. Vandamme, P., Holmes, B., Vancanneyt, M. & 8 other authors ( 1997; ). Occurrence of multiple genomovars of Burkholderia cepacia in cystic fibrosis patients and proposal of Burkholderia multivorans sp. nov. Int J Syst Bacteriol 47, 1188–1200.[CrossRef]
    [Google Scholar]
  31. Vandamme, P., Mahenthiralingam, E., Holmes, B., Coenye, T., Hoste, B., De Vos, P., Henry, D. & Speert, D. P. ( 2000; ). Identification and population structure of Burkholderia stabilis sp. nov. (formerly Burkholderia cepacia genomovar IV). J Clin Microbiol 38, 1042–1047.
    [Google Scholar]
  32. Vandamme, P., Holmes, B., Coenye, T., Goris, J., Mahenthiralingam, E., LiPuma, J. J. & Govan, J. R. ( 2003; ). Burkholderia cenocepacia sp. nov. – a new twist to an old story. Res Microbiol 154, 91–96.[CrossRef]
    [Google Scholar]
  33. Vermis, K., Coenye, T., Mahenthiralingam, E., Nelis, H. J. & Vandamme, P. ( 2002a; ). Evaluation of species-specific recA-based PCR tests for genomovar level identification within the Burkholderia cepacia complex. J Med Microbiol 51, 937–940.
    [Google Scholar]
  34. Vermis, K., Vandekerckhove, C., Nelis, H. J. & Vandamme, P. A. ( 2002b; ). Evaluation of restriction fragment length polymorphism analysis of 16S rDNA as a tool for genomovar characterisation within the Burkholderia cepacia complex. FEMS Microbiol Lett 214, 1–5.[CrossRef]
    [Google Scholar]
  35. Vermis, K., Coenye, T., LiPuma, J. J., Mahenthiralingam, E., Nelis, H. J. & Vandamme, P. ( 2004; ). Proposal to accommodate Burkholderia cepacia genomovar VI as Burkholderia dolosa sp. nov. Int J Syst Evol Microbiol 54, 689–691.[CrossRef]
    [Google Scholar]
  36. Whitby, P. W., Carter, K. B., Hatter, K. L., LiPuma, J. J. & Stull, T. L. ( 2000; ). Identification of members of the Burkholderia cepacia complex by species-specific PCR. J Clin Microbiol 38, 2962–2965.
    [Google Scholar]
  37. Woods, C. W., Bressler, A. M., LiPuma, J. J., Alexander, B. D., Clements, D. A., Weber, D. J., Moore, C. M., Reller, L. B. & Kaye, K. S. ( 2004; ). Virulence associated with outbreak-related strains of Burkholderia cepacia complex among a cohort of patients with bacteremia. Clin Infect Dis 38, 1243–1250.[CrossRef]
    [Google Scholar]
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