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DETECÇAO SIMULTANEA DOS GENES MECA E ILES-2 EM STAPHYLOCOCCUS COAGULASE NEGATIVOS ISOLADOS DE HOSPITAIS BRASILEIROS ATRAVES DA TECNICA DE PCR-MULTIPLEX
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O estudo mostrou que o PCR-multiplex é necessário para determinar taxas mais precisas de resistência a oxacilina e a mupirocina em SCN, podendo ajudar no controle e prevenção de infecções estafilocócicas no Brasil.
regina9.jpg Autor:
Kátia Regina Netto Dos Santos,
Columnista Experto de SIIC

Institución:
Instituto de Microbiología Universidade Federal do Rio de Janeiro Rio de Janeiro, Brasil


Artículos publicados por Kátia Regina Netto Dos Santos,
Coautores
Rosana Barreto Rocha Ferreira*  Ana Paula Ferreira Nunes.*  Valeria Miguelote Kokis.*  Natascha Krepsky.*  Leila Souza Fonseca.**  Marcia Giambiagi de Marval.*** 
Mestre em MicrobiologiaInstituto de Microbiologia da Universidade Federal do Rio de Janeiro*
Professor Adjunto; Doutora em MicrobiologiaInstituto de Microbiologia da Universidade Federal do Rio de Janeiro**
Professora adjunto; Doutora em Microbiologia Instituto de Microbiologia da Universidade Federal do Rio de Janeiro***
Recepción del artículo
12 de Abril, 2004
Aprobación
8 de Junio, 2004
Primera edición
4 de Agosto, 2004

Resumen
A susceptibilidade a oxacilina (Oxa) e a mupirocina (Mup) foi analisada por diferentes métodos em amostras hospitalares de SCN. Entre 112 amostras, 69 (61.6%) foram OxaR pelo teste de difusão do disco (DD) e 72 (64.2%) cresceram no agar triagem com 6 μg/ml de oxacilina. S. epidermidis e S. haemolyticus apresentaram taxas elevadas de resistência a oxacilina, 75.4% e 96.1%, respectivamente. Vinte e quatro (21.4%) amostras foram MupR pelo DD. A detecção dos genes mecA e ileS-2 por PCR-multiplex mostrou que 72 (64.2%) dos SCN possuíam o gene mecA e 16 (14.3%) apresentavam o gene ileS-2. Quinze dessas amostras apresentaram os dois genes simultaneamente. As amostras contendo o gene ileS-2 apresentavam CMI > 1 024 μg/mL pelo teste-E, enquanto resistência a baixos níveis de mupirocina (CMIs de 12–16 μg/mL) foi observada entre as amostras sem o ileS-2. A resistência a altos e baixos níveis de mupirocina não pode ser diferenciada pelo DD. A análise das amostras de S. epidermidis MupR por PFGE mostrou um perfil genotípico prevalente (80.9%) nos hospitais do Rio de Janeiro. O estudo mostrou que o PCR-multiplex é necessário para determinar taxas mais precisas de resistência a Oxa e a Mup em SCN, podendo ajudar no controle e prevenção de infecções estafilocócicas no Brasil.

Palabras clave
Staphylococcus coagulase negativos, resistência a mupirocina, resistência a oxacilina, PCR-multiplex, Eletroforese em gel de campo pulsado (PFGE)


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Abstract
Susceptibility to oxacillin and mupirocin was analyzed by different testing methods in clinical CNS isolates. Among 112 CNS strains, 69 (61.6%) were OxaR by the disk diffusion test (DD) and 72 (64.2%) grew on the oxacillin agar screen plate. S. epidermidis and S. haemolyticus presented high rates of oxacillin resistance (75.4 % and 96.1 %, respectively). Twenty four (21.4%) strains were MupR by the DD. The detection of the mecA and ileS-2 genes, determined by multiplex-PCR, showed that 72 (64.2%) CNS strains possessed the mecA gene and 16 (14.3%) possessed the ileS-2. Fifteen of these strains presented the two resistance genes simultaneously. The isolates containing the ileS-2 presented a MIC > 1 024 μg/mL in the E-test, while low-level mupirocin resistance (MICs of 12–16 μg/mL) was observed in strains without ileS-2. The resistances to high and low levels of mupirocin could not be distinguished by DD. The analysis of the MupR S. epidermidis strains by PFGE showed a prevalent profile (80.9%) in hospitals in Rio de Janeiro. This report showed that the multiplex-PCR is necessary to determine accurate rates of resistance to oxacillin and mupirocin in CNS, and will can help in the staphylococcal infections prevention and control policies in Brazil.

Key words
Coagulase-negative staphylococci; mupirocin r


Clasificación en siicsalud
Artículos originales > Expertos de Iberoamérica >
página   www.siicsalud.com/des/expertocompleto.php/

Especialidades
Principal: Diagnóstico por LaboratorioInfectología
  Relacionadas: EpidemiologíaMedicina InternaSalud Pública

Enviar correspondencia a:
Netto dos Santos, Kátia Regina
Bibliografía del artículo
  1. Rupp ME & Archer GL. Coagulase-negative staphylococci: pathogens associated with medical progress. Clinical and Infectious Diseases 1994; 19: 231–245.
  2. Diekema DJ, Pfaller MA, Jones RN, et al. Trends in antimicrobial susceptibility of bacterial pathogens isolated from patients with bloodstream infections in the USA, Canada and Latin America. SENTRY Participants Group. Int. J. Antim. Agents.2000, 13: 257-271.
  3. Boyce JM. Preventing staphylococcal infection by eradicating nasal carriage of Staphylococcus aureus: proceeding with caution. Infectious Control and Hospital Epidemiology 1996; 17: 775–779.
  4. Cookson BD. The emergence of mupirocin resistance: a challenge to infection control and antibiotic prescribing practice. Journal of Antimicrobial Chemotherapy 1990; 41: 11–18.
  5. Santos KRN, Fonseca LS & Gontijo Filho PP. Emergence of high-level mupirocin resistance in methicillin-resistant Staphylococcus aureus isolated from Brazilian university hospitals. Infectious Control and Hospital Epidemiology 1996; 17: 24–27.
  6. Gilbart J, Perry CR & Slocombe B. High-level mupirocin resistance in Staphylococcus aureus: evidence of two distinct isoleucyltRNA synthetases. Antimicrobial Agents and Chemotherapy 1993; 37:32–38.
  7. Bastos MCF, Mondino PJJ, Azevedo MLB, et al. Molecular characterization and transfer among Staphylococcus strains of a plasmid conferring highlevel esistance to mupirocin. European Journal of Clinical and Infectious Disease 1999; 18: 393–398.
  8. Schmitz FJ, Lindenlauf E, Hofmann B, et al. The prevalence of low-and high-level mupirocin resistance in staphylococci from 19 European hospitals. Journal of Antimicrobial Chemotherapy 1998; 42: 489–495.
  9. Nunes ELC, Santos KRN, Mondino PJJ, et al. Detection of ileS-2 gene encoding mupirocin resistance in methicillin-resistant Staphylococcus aureus by multiplex PCR. Diagn Microbiology and Infectious Disease 1999; 34: 77–71.
  10. Kloos WE & Lambe DWJR. Staphylococcus and Micrococcus. In P. R. Murray, E. J. Baron, M. A. Pfaller, F. C. Tenover, & R. H. Yolken (Eds.), Manual of clinical microbiology, 6th edn. 1995; Washington, DC: American Society for Microbiology.
  11. National Committee For Clinical Laboratory Standards. Performance standards for antimicrobial disk susceptibility tests, 2000; 7th edn. Villanova, PA: NCCLS. Approved standard: M2–A7.
  12. National Committee For Clinical Laboratory Standards. Performance standards for antimicrobial susceptibility testing. 2001 Eleventh in 205–212 formational supplement. Villanova, PA: NCCLS. Approved standard: M100–S11.
  13. Fuchs PC, Jones RDN & Barry AL. Interpretive criteria for disk diffusion susceptibility testing of mupirocin, a topical antibiotic. Journal of Clinical Microbiology 1990; 28: 608–609.
  14. Jorgensen M, Givney R, Pegler M, et al. Typing multidrug-resistant Staphylococcus aureus: conflicting, epidemiological data produced by genotypic and phenotypic methods clarified by phylogenetic analysis. Journal of Clinical Microbiology 1993; 34: 398–403.
  15. Tenover FC, Arbeit RD, Goering RV, et al. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. Journal of Clinical Microbiology 1995; 33: 2233–2239.
  16. Del'Alamo L, Cereda RF, Tosin I, et al. Antimicrobial susceptibility of coagulase-negative staphylococci and characterization of isolates with reduced susceptibility to glycopeptides. Diagn Microbiology Infectious Disease 1999; 34: 185–191.
  17. Santos KRN, Fonseca LS, Bravo Neto GP, et al. Surgical site infection: rates, etiology and resistance patterns to antimicrobials among strains isolated at Rio de Janeiro University Hospital. Infection 1997; 25: 217–220.
  18. Frebourg NB, Nouet D, Leme´e L, et al. Comparison of ATB staph, rapid ATB staph, vitek, and E-test methods for detection of oxacillin heteroresistance in staphylococci possessing mecA. Journal of Clinical Microbiology 1998; 36: 52–57.
  19. Kloos WE & Bannerman TL. Update on clinical significance of coagulase-negative staphylococci. Clinical Microbiology Reviews 1994; 7: 117–140.
  20. Hudson IRB. The efficacy of intranasal mupirocin in the prevention of staphylococcal infections: a review of recent experience. Journal of Hospital Infection 1994; 27: 81–98.
  21. Leski TA, Gniadkowski M, Skoczynska A, et al. Outbreak of mupirocin-resistant staphylococci in a hospital in Warsaw, Poland, due to plasmid transmission and clonal spread of several strains. Journal of Clinical Microbiology 1999; 37: 2781–2788.
  22. Eltringham I. Mupirocin resistance and methicillin-resistant Staphylococcus aureus (MRSA). Journal of Hospital Infection 1997; 35: 1–8.
  23. Santos KRN, Teixeira LM, Leal GS, et al. DNA typing of methicillin-resistant Staphylococcus aureus: isolates and factors associated with nosocomial acquisition in two Brazilian university hospitals. Journal of Medical and Microbiology 1999; 48: 17–23.
  24. Nunes APF. M.Sc. thesis. Federal University of Rio de Janeiro, 2000; Rio de Janeiro, Brazil.

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Especialidad principal:
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Relacionadas:
 Epidemiología
 Medicina Interna
 Salud Pública
 
 
 
 
 
 
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