Genetic Profiling of Pseudomonas aeruginosa Isolates from Iranian Patients with Cystic Fibrosis Using RAPD-PCR and PFGE | ||
| Iranian Journal of Basic Medical Sciences | ||
| مقاله 1، دوره 12، شماره 3، مهر 2009، صفحه 126-132 اصل مقاله (295 K) | ||
| نوع مقاله: Original Article | ||
| شناسه دیجیتال (DOI): 10.22038/ijbms.2009.5164 | ||
| نویسندگان | ||
| Fereshteh Eftekhar* 1؛ Nazanin Hosseinkhan1؛ Ahmad Asgharzadeh2؛ Ahmad Tabatabaii3 | ||
| 1Department of Microbiology, Faculty of Biological Sciences, Shahid Beheshti University, GC, Tehran, Iran | ||
| 2Soil and Water Research Institute, Tehran, Iran | ||
| 3Medical School, Shahid Beheshti University, Tehran, Iran | ||
| چکیده | ||
| Objective(s) Pseudomonas aeruginosa is the most important cause of chronic lung infections and death in patients with cystic fibrosis. Determining the distribution of specific strains within patient populations is important in order to examine the epidemiology of the disease and the possibility of cross infection among patients. Materials and Methods Forty six Iranian patients with cystic fibrosis were studied for colonization with P. aeruginosa. Colony phenotype was recorded and antibiotic susceptibility to '' antibiotics was determined using the disc diffusion method. Genetic fingerprinting was carried out by RAPD-PCR and by PFGE. Results Forty five P. aeruginosa isolates were recovered from 31 patients including sequential cultures from 9 subjects. The rate of colonization increased with age. All isolates were susceptible to tobramycin and ciprofloxacin, 97.8% were sensitive to amikacin and piperacillin, 93.3% to gentamycin, 91.1% to ticarcillin, 86.7% to colistin, 80% to carbenicillin, 48.9% to cefotaxime, 26.7% to imipenem and 11.1% to ceftazidime. Genetic fingerprinting showed similar distribution profiles for RAPD-PCR and PFGE and the majority of the isolates had unique fingerprints. Conclusion No relationship was observed between the obtained genotypes and antibiotic susceptibility profiles and common predominant virulent clones were not found among the isolates. | ||
| کلیدواژهها | ||
| Cystic fibrosis؛ Genetic fingerprinting؛ PFGE؛ Psuedomonas aeruginosa؛ RAPD-PCR | ||
| مراجع | ||
|
1.Govan JRW, Deretic V. Microbial pathogenesis in cystic fibrosis: Mucoid Pseudomonas aeruginosa and Burkholderia cepacia .Microbiol Rev 1996; 60:539-574.
2.Hoiby N, Doring G, Schiotz PO. Microbiology of lung infections in cystic fibrosis. Acta Paediatr Scand Suppl 1986; 301: 3-54.
3.Koch C, Hoiby N. Pathogenesis of cystic fibrosis. Lancet 1993; 341:1065-1069.
4.Burns MW, May JR. Bacterial precipitins in serum of patients with cystic fibrosis. Lancet I 1968; 270-272.
5.Diaz F, Mosovich LL, Netter E. Serogroups of Pseudomonas aeruginosa and the immune response of patients with cystic fibrosis. J Infect Dis 1970; 121:269-274.
6.Speert DP, Farmer SW, Campbell ME, Musser JM, Selander RK, Kuo S. Conversion of Pseudomonas aeruginosa to mucoid phenotype characteristic of strains from patients with cystic fibrosis. J Clin Microbiol 1990; 28:188-194.
7.Adams C, Morris-Quinn M, McConnell F, West J, Lucey B, Shortt C, et al. Epidemiology and clinical impact of Pseudomonas aeruginosa infection in cystic fibrosis using AP-PCR fingerprinting. J Infect 1998; 37:151-158.
8.Brokopp CD, Farmer JJ. Typing methods for Pseudomonas aeruginosa. In: Dogget EG, editor. Pseudomonas aeruginosa: Clinical manifestations of infection and current therapy. NY: Academic Press Inc; 1979. p.89-133.
9.Hancock RE, Mutharia LM, Darveau RP, Darveau RP, Speert DP, Pier GB. Pseudomonas aeruginosa isolates from patients with cystic fibrosis: A class of serum sensitive non-typable strains deficient in lipopolysaccharide side chains. Infect Immun 1983; 2:170-177.
10.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; 33:528-534.
11.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; 155:119- 126.
12.Ojeniyi B, Petersen US, Hoiby N. Comparison of genome fingerprinting with conventional typing methods used on Pseudomonas aeruginosa isolates from cystic fibrosis patients. APMIS 1993; 101:168-175.
13.Welsh J, McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 1990; 18:7213-7218.
14.Mahenthiralingam E, Campbell ME, Foster J, Lam JS, Speert DP. Random amplified polymorphic typing of Pseudomonas aeruginosa isolates recovered from patients with cystic fbrosis. J Clin Microbiol 1996; 34:1129-1135.
15.Kersulyte D, Struelens MJ, Delpano A, Berg DE. Comparison of arbitrarily primed PCR and macro restriction (pulse-field gel electrophoresis) typing methods of Pseudomonas aeruginosa strains from cystic fbrosis patients. J Clin Microbiol 1995; 33:2216-2219.
16.National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial disk susceptibility tests. 6th ed. Approved standard M2-A6 Villlanoca, PA, USA; 1997.
17.Richardson AE, Viccars LA, Watson JM, Gibson AH. Differentiation of Rhizobium strains using the polymerase chain reaction with random and directed primer. Soil Biol Biochem 1995; 27:515-524.
18.Lightfoot J, Lam JS. Chromosomal mapping, expression and synthesis of lipopolysaccaride in Pseudomonas aeruginosa: a role for guanosine diphospho (GDP)-D- mannose. Mol Microbiol 1993; 8:771-782.
19.Romling U, Grothues D, Bautsch W, Tummler B. A physical genome map of Pseudomonas aeruginosa PAO. EMBO J 1989; 8:4081-4089.
20.Eftekhar F, Rostamizadeh F, Khodadad A, Henry D, Speert DP. Isolation and characterization of Pseudomonas aeruginosa from Iranian patients with cystic fibrosis using RAPD-PCR. Iran J Biotech 2003; 1:95-100.
21.Ballestero S, Fernandez-Rodriguez A, Villaverde R, Escobar H, Perez-Diaz JC, Baquero F. Carbapenem resistance in Pseudomonas aeruginosa from cystic fibrosis patients. J Antimicrob Chemother 1996; 38:39-45.
22.Ernst-Jurgen W, Witte W. Instability of in vitro resistance to imipenem in Pseudomonas aeruginosa. J Basic Microbiol 1990; 30:617-622.
23.Kim IS, Lee NY, Ki CS, Oh WS, Peck KR, Song JH. Increasing prevalence of imipenem-resistant Pseudomonas aeruginosa and molecular typing of metallo-p-lactamase producers in a Korean hospital. Microb Drug Resist 2005; 11:355-359.
24.Vosahlikova S, Drevinek P, Cinek O, Pohunek P, Maixnerova M, Urbaskova P, et al. High genotypic diversity of Pseudomonas aeruginosa strains isolated from patients with cystic fibrosis in the Czech Republic. Res Microbiol 2007; 158:324-329.
25.Schmid J, Ling LJ, Leung JLS, Zhang N, Kolbe J, Wesley AW, et al. Pseudomonas aeruginosa transmission is infrequent in New Zealand cystic fibrosis clinics. Eur Respir J 2008; 32:1583-7590.
26.Scott FW, Pitt TL. Identification and characterization of transmissible Pseudomonas aeruginosa strains in cystic fibrosis patients in England and Wales. J Med Microbiol 2004; 53:609-515.
27.Hogardt M, Hoboth C, Schmoldt S, Henke C, Bader L, Heesemann J. Stage-specific adaptation of hypermutable Pseudomonas aeruginosa isolates during chronic pulmonary infection in patients with cystic fibrosis. J Infect Dis 2007; 195:70-80.
28.Keena DT, Doherty CD, Foweraker J, Macaskill L, Barcus VA, Govan JR. Hypermutability in environmental Pseudomonas aeruginosa and in populations causing pulmonary infection in individuals with cystic fibrosis. Microbiology 2007; 153:1852-1859.
29.Tingpej P, Smith L, Rose B, Zhu H, Conibear T, Al Nassafi K, et al. Phenotypic characterization of clonal and nonclonal Pseudomonas aeruginosa strains isolated from lungs of adults with cystic fibrosis. J Clin Microbiol 2007; 45:1697-1704. | ||
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