Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. 1996 May; 34(5): 1129–1135.
PMCID: PMC228968

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


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.

Full Text

The Full Text of this article is available as a PDF (501K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Akopyanz N, Bukanov NO, Westblom TU, Kresovich S, Berg DE. DNA diversity among clinical isolates of Helicobacter pylori detected by PCR-based RAPD fingerprinting. Nucleic Acids Res. 1992 Oct 11;20(19):5137–5142. [PMC free article] [PubMed]
  • Fegan M, Francis P, Hayward AC, Fuerst JA. Heterogeneity, persistence, and distribution of Pseudomonas aeruginosa genotypes in cystic fibrosis patients. J Clin Microbiol. 1991 Oct;29(10):2151–2157. [PMC free article] [PubMed]
  • Grundmann H, Schneider C, Hartung D, Daschner FD, Pitt TL. Discriminatory power of three DNA-based typing techniques for Pseudomonas aeruginosa. J Clin Microbiol. 1995 Mar;33(3):528–534. [PMC free article] [PubMed]
  • Hancock RE, Mutharia LM, Chan L, Darveau RP, Speert DP, Pier GB. 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. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Hoogkamp-Korstanje JA, Meis JF, Kissing J, van der Laag J, Melchers WJ. 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. [PMC free article] [PubMed]
  • Kersulyte D, Struelens MJ, Deplano A, Berg DE. 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. [PMC free article] [PubMed]
  • Lightfoot J, Lam JS. 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. [PubMed]
  • Liu SL, Hessel A, Sanderson KE. 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. [PMC free article] [PubMed]
  • Mahenthiralingam E, Campbell ME, Speert DP. Nonmotility and phagocytic resistance of Pseudomonas aeruginosa isolates from chronically colonized patients with cystic fibrosis. Infect Immun. 1994 Feb;62(2):596–605. [PMC free article] [PubMed]
  • Mahenthiralingam E, Speert DP. 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. [PMC free article] [PubMed]
  • Ogle JW, Janda JM, Woods DE, Vasil ML. Characterization and use of a DNA probe as an epidemiological marker for Pseudomonas aeruginosa. J Infect Dis. 1987 Jan;155(1):119–126. [PubMed]
  • Pier GB, Meluleni G, Neuger E. A murine model of chronic mucosal colonization by Pseudomonas aeruginosa. Infect Immun. 1992 Nov;60(11):4768–4776. [PMC free article] [PubMed]
  • Ramphal R, Koo L, Ishimoto KS, Totten PA, Lara JC, Lory S. Adhesion of Pseudomonas aeruginosa pilin-deficient mutants to mucin. Infect Immun. 1991 Apr;59(4):1307–1311. [PMC free article] [PubMed]
  • 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. [PubMed]
  • 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. [PubMed]
  • Sokol PA, Luan MZ, Storey DG, 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. [PMC free article] [PubMed]
  • Speert DP, Campbell ME. Hospital epidemiology of Pseudomonas aeruginosa from patients with cystic fibrosis. J Hosp Infect. 1987 Jan;9(1):11–21. [PubMed]
  • Speert DP, Campbell ME, Farmer SW, Volpel K, Joffe AM, Paranchych W. Use of a pilin gene probe to study molecular epidemiology of Pseudomonas aeruginosa. J Clin Microbiol. 1989 Nov;27(11):2589–2593. [PMC free article] [PubMed]
  • Struelens MJ, 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. [PMC free article] [PubMed]
  • Tang H, Kays M, Prince A. Role of Pseudomonas aeruginosa pili in acute pulmonary infection. Infect Immun. 1995 Apr;63(4):1278–1285. [PMC free article] [PubMed]
  • Totten PA, Lara JC, 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. [PMC free article] [PubMed]
  • Welsh J, McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 1990 Dec 25;18(24):7213–7218. [PMC free article] [PubMed]
  • Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. [PMC free article] [PubMed]
  • Wolz C, Kiosz G, Ogle JW, Vasil ML, 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. [PMC free article] [PubMed]
  • Woods DE, Sokol PA, Bryan LE, Storey DG, Mattingly SJ, Vogel HJ, Ceri H. In vivo regulation of virulence in Pseudomonas aeruginosa associated with genetic rearrangement. J Infect Dis. 1991 Jan;163(1):143–149. [PubMed]

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


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...