Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Apr;35(4):808–816. doi: 10.1128/jcm.35.4.808-816.1997

Identification and characterization of a novel DNA marker associated with epidemic Burkholderia cepacia strains recovered from patients with cystic fibrosis.

E Mahenthiralingam 1, D A Simpson 1, D P Speert 1
PMCID: PMC229681  PMID: 9157133

Abstract

Burkholderia cepacia is a problematic pathogen that may spread among patients with cystic fibrosis (CF). One highly infectious CF strain that causes epidemics in both the United Kingdom and eastern Canada has been shown to possess both the cable pilin subunit gene (cblA) and a unique combination of insertion sequences. However, no genetic markers linking this strain type with other types epidemic at various centers have been identified. Using a randomly amplified polymorphic DNA (RAPD) typing scheme, we identified an apparently conserved 1.4-kb fragment in the DNA fingerprint of epidemic B. cepacia strains. Conservation of the DNA marker among epidemic strains was demonstrated by Southern hybridization, and its prevalence was assessed in a collection of chromosomal DNAs extracted from 627 isolates representative of 132 RAPD-defined B. cepacia strain types. The marker was specifically associated with seven epidemic CF strains, was absent among nonepidemic strains infecting individual patients with CF, and rare among strains recovered from the natural environment. Only one of the seven epidemic CF strain types possessed DNA homologous to cblA. The RAPD marker was designated the "B. cepacia epidemic strain marker" (BCESM). Sequence analysis of chromosomal DNA corresponding to the 1.4-kb RAPD marker revealed the presence of a putative open reading frame (ORF) with significant homology to several negative transcriptional regulators; the ORF was designated the "epidemic strain marker regulator," or esmR. The BCESM DNA is the first genetic marker that has been identified to be specifically associated with and conserved among several epidemic B. cepacia strains which infect multiple patients with CF.

Full Text

The Full Text of this article is available as a PDF (922.6 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Blanco C., Ritzenthaler P., Kolb A. The regulatory region of the uxuAB operon in Escherichia coli K12. Mol Gen Genet. 1986 Jan;202(1):112–119. doi: 10.1007/BF00330526. [DOI] [PubMed] [Google Scholar]
  3. Burland V., Plunkett G., 3rd, Sofia H. J., Daniels D. L., Blattner F. R. Analysis of the Escherichia coli genome VI: DNA sequence of the region from 92.8 through 100 minutes. Nucleic Acids Res. 1995 Jun 25;23(12):2105–2119. doi: 10.1093/nar/23.12.2105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burns J. L., Wadsworth C. D., Barry J. J., Goodall C. P. Nucleotide sequence analysis of a gene from Burkholderia (Pseudomonas) cepacia encoding an outer membrane lipoprotein involved in multiple antibiotic resistance. Antimicrob Agents Chemother. 1996 Feb;40(2):307–313. doi: 10.1128/aac.40.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Butler S. L., Doherty C. J., Hughes J. E., Nelson J. W., Govan J. R. Burkholderia cepacia and cystic fibrosis: do natural environments present a potential hazard? J Clin Microbiol. 1995 Apr;33(4):1001–1004. doi: 10.1128/jcm.33.4.1001-1004.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cervin M. A., Simpson D. A., Smith A. L., Lory S. Differences in eucaryotic cell binding of Pseudomonas. Microb Pathog. 1994 Nov;17(5):291–299. doi: 10.1006/mpat.1994.1075. [DOI] [PubMed] [Google Scholar]
  7. Cheng H. P., Lessie T. G. Multiple replicons constituting the genome of Pseudomonas cepacia 17616. J Bacteriol. 1994 Jul;176(13):4034–4042. doi: 10.1128/jb.176.13.4034-4042.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dong J. M., Taylor J. S., Latour D. J., Iuchi S., Lin E. C. Three overlapping lct genes involved in L-lactate utilization by Escherichia coli. J Bacteriol. 1993 Oct;175(20):6671–6678. doi: 10.1128/jb.175.20.6671-6678.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gaffney T. D., Lessie T. G. Insertion-sequence-dependent rearrangements of Pseudomonas cepacia plasmid pTGL1. J Bacteriol. 1987 Jan;169(1):224–230. doi: 10.1128/jb.169.1.224-230.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldstein R., Sun L., Jiang R. Z., Sajjan U., Forstner J. F., Campanelli C. Structurally variant classes of pilus appendage fibers coexpressed from Burkholderia (Pseudomonas) cepacia. J Bacteriol. 1995 Feb;177(4):1039–1052. doi: 10.1128/jb.177.4.1039-1052.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Govan J. R., Brown P. H., Maddison J., Doherty C. J., Nelson J. W., Dodd M., Greening A. P., Webb A. K. Evidence for transmission of Pseudomonas cepacia by social contact in cystic fibrosis. Lancet. 1993 Jul 3;342(8862):15–19. doi: 10.1016/0140-6736(93)91881-l. [DOI] [PubMed] [Google Scholar]
  12. Haydon D. J., Guest J. R. A new family of bacterial regulatory proteins. FEMS Microbiol Lett. 1991 Apr 15;63(2-3):291–295. doi: 10.1016/0378-1097(91)90101-f. [DOI] [PubMed] [Google Scholar]
  13. Henikoff S., Henikoff J. G. Protein family classification based on searching a database of blocks. Genomics. 1994 Jan 1;19(1):97–107. doi: 10.1006/geno.1994.1018. [DOI] [PubMed] [Google Scholar]
  14. Hutchinson G. R., Parker S., Pryor J. A., Duncan-Skingle F., Hoffman P. N., Hodson M. E., Kaufmann M. E., Pitt T. L. Home-use nebulizers: a potential primary source of Burkholderia cepacia and other colistin-resistant, gram-negative bacteria in patients with cystic fibrosis. J Clin Microbiol. 1996 Mar;34(3):584–587. doi: 10.1128/jcm.34.3.584-587.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Høiby N. Isolation and treatment of cystic fibrosis patients with lung infections caused by Pseudomonas (Burkholderia) cepacia and multiresistant Pseudomonas aeruginosa. Neth J Med. 1995 Jun;46(6):280–287. doi: 10.1016/0300-2977(95)00020-n. [DOI] [PubMed] [Google Scholar]
  16. Johnson W. M., Tyler S. D., Rozee K. R. Linkage analysis of geographic and clinical clusters in Pseudomonas cepacia infections by multilocus enzyme electrophoresis and ribotyping. J Clin Microbiol. 1994 Apr;32(4):924–930. doi: 10.1128/jcm.32.4.924-930.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. LiPuma J. J., Dasen S. E., Nielson D. W., Stern R. C., Stull T. L. Person-to-person transmission of Pseudomonas cepacia between patients with cystic fibrosis. Lancet. 1990 Nov 3;336(8723):1094–1096. doi: 10.1016/0140-6736(90)92571-x. [DOI] [PubMed] [Google Scholar]
  18. LiPuma J. J., Mortensen J. E., Dasen S. E., Edlind T. D., Schidlow D. V., Burns J. L., Stull T. L. Ribotype analysis of Pseudomonas cepacia from cystic fibrosis treatment centers. J Pediatr. 1988 Nov;113(5):859–862. doi: 10.1016/s0022-3476(88)80018-0. [DOI] [PubMed] [Google Scholar]
  19. Mahenthiralingam E., Campbell M. E., Foster J., Lam J. S., Speert D. P. Random amplified polymorphic DNA typing of Pseudomonas aeruginosa isolates recovered from patients with cystic fibrosis. J Clin Microbiol. 1996 May;34(5):1129–1135. doi: 10.1128/jcm.34.5.1129-1135.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mahenthiralingam E., Campbell M. E., Henry D. A., Speert D. P. Epidemiology of Burkholderia cepacia infection in patients with cystic fibrosis: analysis by randomly amplified polymorphic DNA fingerprinting. J Clin Microbiol. 1996 Dec;34(12):2914–2920. doi: 10.1128/jcm.34.12.2914-2920.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nelson J. W., Butler S. L., Krieg D., Govan J. R. Virulence factors of Burkholderia cepacia. FEMS Immunol Med Microbiol. 1994 Feb;8(2):89–97. doi: 10.1111/j.1574-695X.1994.tb00430.x. [DOI] [PubMed] [Google Scholar]
  22. Rodley P. D., Römling U., Tümmler B. A physical genome map of the Burkholderia cepacia type strain. Mol Microbiol. 1995 Jul;17(1):57–67. doi: 10.1111/j.1365-2958.1995.mmi_17010057.x. [DOI] [PubMed] [Google Scholar]
  23. Sajjan S. U., Forstner J. F. Identification of the mucin-binding adhesin of Pseudomonas cepacia isolated from patients with cystic fibrosis. Infect Immun. 1992 Apr;60(4):1434–1440. doi: 10.1128/iai.60.4.1434-1440.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sajjan U. S., Forstner J. F. Role of a 22-kilodalton pilin protein in binding of Pseudomonas cepacia to buccal epithelial cells. Infect Immun. 1993 Aug;61(8):3157–3163. doi: 10.1128/iai.61.8.3157-3163.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sajjan U. S., Sun L., Goldstein R., Forstner J. F. Cable (cbl) type II pili of cystic fibrosis-associated Burkholderia (Pseudomonas) cepacia: nucleotide sequence of the cblA major subunit pilin gene and novel morphology of the assembled appendage fibers. J Bacteriol. 1995 Feb;177(4):1030–1038. doi: 10.1128/jb.177.4.1030-1038.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Speert D. P., Bond M., Woodman R. C., Curnutte J. T. Infection with Pseudomonas cepacia in chronic granulomatous disease: role of nonoxidative killing by neutrophils in host defense. J Infect Dis. 1994 Dec;170(6):1524–1531. doi: 10.1093/infdis/170.6.1524. [DOI] [PubMed] [Google Scholar]
  27. Stephens P. E., Darlison M. G., Lewis H. M., Guest J. R. The pyruvate dehydrogenase complex of Escherichia coli K12. Nucleotide sequence encoding the pyruvate dehydrogenase component. Eur J Biochem. 1983 Jun 1;133(1):155–162. doi: 10.1111/j.1432-1033.1983.tb07441.x. [DOI] [PubMed] [Google Scholar]
  28. Sun L., Jiang R. Z., Steinbach S., Holmes A., Campanelli C., Forstner J., Sajjan U., Tan Y., Riley M., Goldstein R. The emergence of a highly transmissible lineage of cbl+ Pseudomonas (Burkholderia) cepacia causing CF centre epidemics in North America and Britain. Nat Med. 1995 Jul;1(7):661–666. doi: 10.1038/nm0795-661. [DOI] [PubMed] [Google Scholar]
  29. Tablan O. C., Chorba T. L., Schidlow D. V., White J. W., Hardy K. A., Gilligan P. H., Morgan W. M., Carson L. A., Martone W. J., Jason J. M. Pseudomonas cepacia colonization in patients with cystic fibrosis: risk factors and clinical outcome. J Pediatr. 1985 Sep;107(3):382–387. doi: 10.1016/s0022-3476(85)80511-4. [DOI] [PubMed] [Google Scholar]
  30. Tyler S. D., Rozee K. R., Johnson W. M. Identification of IS1356, a new insertion sequence, and its association with IS402 in epidemic strains of Burkholderia cepacia infecting cystic fibrosis patients. J Clin Microbiol. 1996 Jul;34(7):1610–1616. doi: 10.1128/jcm.34.7.1610-1616.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Worley K. C., Wiese B. A., Smith R. F. BEAUTY: an enhanced BLAST-based search tool that integrates multiple biological information resources into sequence similarity search results. Genome Res. 1995 Sep;5(2):173–184. doi: 10.1101/gr.5.2.173. [DOI] [PubMed] [Google Scholar]

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

RESOURCES