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. 1996 May;34(5):1129–1135. doi: 10.1128/jcm.34.5.1129-1135.1996

Random amplified polymorphic DNA typing of Pseudomonas aeruginosa isolates recovered from patients with cystic fibrosis.

E Mahenthiralingam 1, M E Campbell 1, J Foster 1, J S Lam 1, D P Speert 1
PMCID: PMC228968  PMID: 8727889

Abstract

Pseudomonas aeruginosa isolates recovered from chronically colonized patients with cystic fibrosis (CF) are phenotypically different from those collected from other patients or from the environment. To assess whether alterations in motility, mucoidy, and serum susceptibility represented an adaptation to chronic infection or replacement by a new strain, sequential P. aeruginosa isolates of known phenotype collected from 20 CF patients were typed by random amplified polymorphic DNA (RAPD) analysis. A total of 35 RAPD strain types were found among 385 isolates from 20 patients, and only two patients had P. aeruginosa strains of the same RAPD fingerprint. Eight strain pairs representative of the first eight RAPD types were also analyzed by SpeI macrorestriction followed by pulsed-field gel electrophoresis (PFGE); the strain types found by both fingerprinting techniques correlated exactly. In 11 of 20 patients, the RAPD types of serial P. aeruginosa isolates remained stable despite alterations in isolate motility, colonial morphology, and lipopolysaccharide phenotype. However, in isolates collected from one CF patient, a single band change in RAPD fingerprint and CeuI PFGE profile correlated with the appearance of an RpoN mutant phenotype, suggesting that the altered phenotype may have been due to a stable genomic rearrangement. Secretion of mucoid exopolysaccharide, loss of expression of RpoN-dependent surface factors, and acquisition of a serum-susceptible phenotype in P. aeruginosa appear to evolve during chronic colonization in CF patients from specific adaptation to infection rather than from acquisition of new bacterial strains.

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Selected References

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  1. Akopyanz N., Bukanov N. O., Westblom T. U., Kresovich S., Berg D. E. DNA diversity among clinical isolates of Helicobacter pylori detected by PCR-based RAPD fingerprinting. Nucleic Acids Res. 1992 Oct 11;20(19):5137–5142. doi: 10.1093/nar/20.19.5137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Fegan M., Francis P., Hayward A. C., Fuerst J. A. Heterogeneity, persistence, and distribution of Pseudomonas aeruginosa genotypes in cystic fibrosis patients. J Clin Microbiol. 1991 Oct;29(10):2151–2157. doi: 10.1128/jcm.29.10.2151-2157.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Grundmann H., Schneider C., Hartung D., Daschner F. D., Pitt T. L. Discriminatory power of three DNA-based typing techniques for Pseudomonas aeruginosa. J Clin Microbiol. 1995 Mar;33(3):528–534. doi: 10.1128/jcm.33.3.528-534.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hancock R. E., Mutharia L. M., Chan L., Darveau R. P., Speert D. P., Pier G. B. Pseudomonas aeruginosa isolates from patients with cystic fibrosis: a class of serum-sensitive, nontypable strains deficient in lipopolysaccharide O side chains. Infect Immun. 1983 Oct;42(1):170–177. doi: 10.1128/iai.42.1.170-177.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hoogkamp-Korstanje J. A., Meis J. F., Kissing J., van der Laag J., Melchers W. J. Risk of cross-colonization and infection by Pseudomonas aeruginosa in a holiday camp for cystic fibrosis patients. J Clin Microbiol. 1995 Mar;33(3):572–575. doi: 10.1128/jcm.33.3.572-575.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Høiby N., Rosendal K. Epidemiology of Pseudomonas aeruginosa infection in patients treated at a cystic fibrosis centre. Acta Pathol Microbiol Scand B. 1980 Jun;88(3):125–131. doi: 10.1111/j.1699-0463.1980.tb02617.x. [DOI] [PubMed] [Google Scholar]
  7. Kersulyte D., Struelens M. J., Deplano A., Berg D. E. Comparison of arbitrarily primed PCR and macrorestriction (pulsed-field gel electrophoresis) typing of Pseudomonas aeruginosa strains from cystic fibrosis patients. J Clin Microbiol. 1995 Aug;33(8):2216–2219. doi: 10.1128/jcm.33.8.2216-2219.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lightfoot J., Lam J. S. Chromosomal mapping, expression and synthesis of lipopolysaccharide in Pseudomonas aeruginosa: a role for guanosine diphospho (GDP)-D-mannose. Mol Microbiol. 1993 May;8(4):771–782. doi: 10.1111/j.1365-2958.1993.tb01620.x. [DOI] [PubMed] [Google Scholar]
  9. Liu S. L., Hessel A., Sanderson K. E. Genomic mapping with I-Ceu I, an intron-encoded endonuclease specific for genes for ribosomal RNA, in Salmonella spp., Escherichia coli, and other bacteria. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6874–6878. doi: 10.1073/pnas.90.14.6874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mahenthiralingam E., Campbell M. E., Speert D. P. Nonmotility and phagocytic resistance of Pseudomonas aeruginosa isolates from chronically colonized patients with cystic fibrosis. Infect Immun. 1994 Feb;62(2):596–605. doi: 10.1128/iai.62.2.596-605.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mahenthiralingam E., Speert D. P. Nonopsonic phagocytosis of Pseudomonas aeruginosa by macrophages and polymorphonuclear leukocytes requires the presence of the bacterial flagellum. Infect Immun. 1995 Nov;63(11):4519–4523. doi: 10.1128/iai.63.11.4519-4523.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ogle J. W., Janda J. M., Woods D. E., Vasil M. L. Characterization and use of a DNA probe as an epidemiological marker for Pseudomonas aeruginosa. J Infect Dis. 1987 Jan;155(1):119–126. doi: 10.1093/infdis/155.1.119. [DOI] [PubMed] [Google Scholar]
  13. Pier G. B., Meluleni G., Neuger E. A murine model of chronic mucosal colonization by Pseudomonas aeruginosa. Infect Immun. 1992 Nov;60(11):4768–4776. doi: 10.1128/iai.60.11.4768-4776.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ramphal R., Koo L., Ishimoto K. S., Totten P. A., Lara J. C., Lory S. Adhesion of Pseudomonas aeruginosa pilin-deficient mutants to mucin. Infect Immun. 1991 Apr;59(4):1307–1311. doi: 10.1128/iai.59.4.1307-1311.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Römling U., Fiedler B., Bosshammer J., Grothues D., Greipel J., von der Hardt H., Tümmler B. Epidemiology of chronic Pseudomonas aeruginosa infections in cystic fibrosis. J Infect Dis. 1994 Dec;170(6):1616–1621. doi: 10.1093/infdis/170.6.1616. [DOI] [PubMed] [Google Scholar]
  16. Saiman L., Ishimoto K., Lory S., Prince A. The effect of piliation and exoproduct expression on the adherence of Pseudomonas aeruginosa to respiratory epithelial monolayers. J Infect Dis. 1990 Mar;161(3):541–548. doi: 10.1093/infdis/161.3.541. [DOI] [PubMed] [Google Scholar]
  17. Sokol P. A., Luan M. Z., Storey D. G., Thirukkumaran P. Genetic rearrangement associated with in vivo mucoid conversion of Pseudomonas aeruginosa PAO is due to insertion elements. J Bacteriol. 1994 Feb;176(3):553–562. doi: 10.1128/jb.176.3.553-562.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Speert D. P., Campbell M. E., Farmer S. W., Volpel K., Joffe A. M., Paranchych W. Use of a pilin gene probe to study molecular epidemiology of Pseudomonas aeruginosa. J Clin Microbiol. 1989 Nov;27(11):2589–2593. doi: 10.1128/jcm.27.11.2589-2593.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Speert D. P., Campbell M. E. Hospital epidemiology of Pseudomonas aeruginosa from patients with cystic fibrosis. J Hosp Infect. 1987 Jan;9(1):11–21. doi: 10.1016/0195-6701(87)90089-2. [DOI] [PubMed] [Google Scholar]
  20. Struelens M. J., Schwam V., Deplano A., Baran D. Genome macrorestriction analysis of diversity and variability of Pseudomonas aeruginosa strains infecting cystic fibrosis patients. J Clin Microbiol. 1993 Sep;31(9):2320–2326. doi: 10.1128/jcm.31.9.2320-2326.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tang H., Kays M., Prince A. Role of Pseudomonas aeruginosa pili in acute pulmonary infection. Infect Immun. 1995 Apr;63(4):1278–1285. doi: 10.1128/iai.63.4.1278-1285.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Totten P. A., Lara J. C., Lory S. The rpoN gene product of Pseudomonas aeruginosa is required for expression of diverse genes, including the flagellin gene. J Bacteriol. 1990 Jan;172(1):389–396. doi: 10.1128/jb.172.1.389-396.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Welsh J., McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 1990 Dec 25;18(24):7213–7218. doi: 10.1093/nar/18.24.7213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Williams J. G., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. doi: 10.1093/nar/18.22.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wolz C., Kiosz G., Ogle J. W., Vasil M. L., Schaad U., Botzenhart K., Döring G. Pseudomonas aeruginosa cross-colonization and persistence in patients with cystic fibrosis. Use of a DNA probe. Epidemiol Infect. 1989 Apr;102(2):205–214. doi: 10.1017/s0950268800029873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Woods D. E., Sokol P. A., Bryan L. E., Storey D. G., Mattingly S. J., Vogel H. J., Ceri H. In vivo regulation of virulence in Pseudomonas aeruginosa associated with genetic rearrangement. J Infect Dis. 1991 Jan;163(1):143–149. doi: 10.1093/infdis/163.1.143. [DOI] [PubMed] [Google Scholar]

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