2004, Number 2
<< Back Next >>
salud publica mex 2004; 46 (2)
RAPD-PCR characterization of Pseudomonas aeruginosa strains obtained fr om cystic fibr osis patients
Ortiz-Herrera M, Gerónimo-Gallegos A, Cuevas-Schacht F, Pérez-Fernández L, Coria-Jiménez R
Language: Spanish
References: 28
Page: 149-157
PDF size: 71.22 Kb.
ABSTRACT
Objective. To characterize
P aeruginosa strains isolated from bronchoalveolar lavage fluid of cystic fibrosis (CF) patients over a 3 year period.
Material and Methods. A prospective follow-up study was carried out in a population of cystic fibrosis patients. The random amplified polymorphic DNA (RAPD) technique was used to amplify DNA of
P aeruginosa strains isolated from bronchoalveolar lavage fluid samples of five CF patients from the Servicio de Neumología y Cirugía del Tórax del Instituto Nacional de Pediatría (Mexico City Chest Clinic of the National Pediatrics Institute) in Mexico City, between June 1996 and June 2002. Amplification patterns were established for each isolate to accurately identify all strains and to carry out an epidemiological analysis of
P aeruginosa among the selected CF patients.
Results. Eighteen different DNA amplification patterns were defined and used to identify each
P aeruginosa strain isolated from the different bronchoalveolar lavage samples. No correlation was observed between the different
P aeruginosa strain genotypes and mucoid or nonmucoid phenotypes, as strains with different phenotypes showed similar amplification patterns. Several strains with different amplification patterns were identified in samples obtained from the same patient, suggesting coinfection with more than one
P aeruginosa strain. Two siblings with CF shared similar genotypes, suggesting the occurrence of cross-contamination. Similar genotypes of
P aeruginosa strains were isolated throughout the study period.
Conclusion. Genotypic characterization of
P aeruginosa strains in CF patients allows more accurate epidemiological analyses of this important host-agent relationship.
REFERENCES
Kiska DL, Gilligan PH. Pseudomonas. En: Murray PR, Baron EJ, Pfaller MA, Tenover FC, Yolken RH, ed. Manual of clinical microbiology. 7th edition. Washington, DC: ASM Press; 1999:517-525.
Walker TS. Microbiology. Philadelphia (PA): OWB Saunders; 1998: 173-181.
Zwadyk P. Pseudomonas. En: Joklil WK, Willet HP, Amos DB, Wilfert CM Ed. Zinsser Microbiology. 20th edition. California (CA): Appleton & Lange; 1992:576-583.
Salyers AA, Whitt DD. Pseudomonas aeruginosa and related species, a lesson of versatility. En: Salyers AA, Whitt DD, ed. Bacterial pathogenesis, a molecular approach. 2nd edition. Washington, DC: ASM Press; 2002:247-262.
Muñoz-Elías EJ, McKinney JD. Bacterial persistence: Strategies for survival. En: Kaufmann SHE, Sher A, Ahmed R, ed. Immunology of infectious diseases. Washington, DC: ASM Press; 2002:331-355.
Snell JK, Bahner DR, Warren RL. Protease phenotypes of Pseudomonas aeruginosa isolated from patients with cystic fibrosis. J Clin Microbiol 1983;17:55-59.
Wolz C, Kiosz G, Ogle JW, Vasil ML, Schaad U, Botzenhart K et al. Pseudomonas aeruginosa cross-colonization and persistence in patients with cystic fibrosis. Use of a DNA probe. Epidemiol Infect 1989;102:205-214.
Horrevortz AM, Borst J, Puyk RJT, de Ridder R, Dzoljic-Danilovic G, Degener JE et al. Ecology of Pseudomonas aeruginosa in patients with cystic fibrosis. J Med Microbiol 1990:119-124.
Richardson CJL, Burke V, Bailey M, Fisher A. Pyocin typing of Pseudomonas aeruginosa isolates from children with cystic fibrosis. Pathology 1991;23:229-232.
Zierdt CH, Williams RL. Serotyping of Pseudomonas aeruginosa isolates from patients with cystic fibrosis of the pancreas. J Clin Microbiol 1975;1:521-526.
Kelly NM, Tempany E, Falkiner Fr, Fitzgerald MX, O’Boyle C, Keane CT. Does Pseudomonas cross-infection occur between cystic fibrosis patients? Lancet 1982;ii:688-690.
Ogle JW, Janda M, Wood DE, Vasil ML. Characterization and use of a DNA probe as an epidemiological marker for Pseudomonas aeruginosa. J Infect Dis 1987;155:119-126.
Grothues D, Koopman U, von der Hart H, Tümmler B. Genome fingerprinting of Pseudomonas aeruginosa indicates colonization of cystic fibrosis siblings with closely related strains. J Clin Microbiol 1988;26:1973-1977.
Struelen 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;31:2320-2326.
Römling U, Fiedler B, Bobhammer J, Grothues D, Geipel J, von der Hart H et al. Epidemiology of chronic Pseudomonas aeruginosa infections in cystic fibrosis. J Infect Dis 1994;170:1616-1621.
Zembrzuska-Sadkowska E, Sneum M, Ojeniyi B, Heiden L, HDiby N. Epidemiology of Pseudomonas aeruginosa infection and the role of contamination of the environment in the Danish cystic fibrosis center. J Hosp Infect 1995;29:1-7.
The International Pseudomonas aeruginosa Typing Study Group. A multicenter comparison of methods for typing strains of Pseudomonas aeruginosa predominantly from patients with cystic fibrosis. J Infect Dis 1994;169:134-142.
Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 1990;18:6531-6535.
Welsh J, McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acid Res 1990;18:7213-7218.
Bingen EH, Weber M, Derelle J, Brahimi N, Lambert-Zechovsky NY Vidailhet M et al. Arbitrarily primed polymerase chain reaction as a rapid method to differentiate crossed from independent Pseudomonas cepacia infections in cystic fibrosis patients. J Clin Microbiol 1993;31:2589-2593.
Bukanov N, Nathan Ravi V, Miller D, Srivastava K, Berg DE. Pseudomonas aeruginosa corneal ulcer isolates distinguished using the arbitrarily primed PCR DNA fingerprinting method. Curr Eye Res 1994;13:783-790.
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;33:2216-2219.
Mahenthiralingam E, Campbell ME, Foster J, Lam JS, Speert DP. Random amplified polymorphic DNA typing of Pseudomonas aeruginosa isolates recovered from patients with cystic fibrosis. J Clin Microbiol 1996;34:1129-1135.
Renders NHM, Sijmons MAF, van Belkum A, Overbeek SE, Mouton JW, Verbrugh HA. Exchange of Pseudomonas aeruginosa strains among cystic fibrosis siblings. Res Microbiol 1997;148:447-454.
Hernández J, Ferrús MA, Hernández M, Owen RJ. Arbitrary primed fingerprinting and serotyping of clinical Pseudomonas aeruginosa strains. FEMS Immunol Med Microbiol 1997;17:37-47.
Adams C, Morris-Quin M, McConnell F, West J, Lucey B, Short C et al. Epidemiology and clinical impact of Pseudomonas aeruginosa infection in cystic fibrosis using AP-PCR fingerprinting. J Infect 1998;37:151-158.
Cuevas Schacht F, Banegas-Matamoros J, Sosa de Martínez C, Coria-Jiménez VR, Pérez-Fernández L, Gerónimo-Gallegos A et al. Identificación de Pseudomonas aeruginosa en pacientes pediátricos con fibrosis quística. Cultivo de expectoración vs lavado broncoalveolar. Acta Pediatr Mex 2001;22:419-423.
Schonian G, Grase Y, Meusel O, Bucholz P, Presber W, Mitchell G. Methods in DNA amplification. Nueva York: Plenum Press; 1994.