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Instituto Nacional de Salud Pública

2004, Number 6

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salud publica mex 2004; 46 (6)

Characterization of SHV-5 β-lactamase-producing Klebsiella pneumoniae in an Intensive Care Unit

Andrade V, Silva J

Language: Spanish
References: 46
Page: 524-528
PDF size: 159.13 Kb.


ABSTRACT

Objective. To perform the molecular characterization of Klebsiella pneumoniae isolates from pediatric patients and health care workers at the intensive care unit of a tertiary care hospital in Mexico City. Material and Methods. Fifteen Klebsiella pneumoniae isolates collected during an outbreak in June 1996 were analyzed; eight were from patients and seven from health care workers of Mexico’s Children’s Hospital. Characterization of isolates was carried out by pulsed field gel electrophoresis (PFGE), random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) and serotyping, β-Lactamase isoelectric focusing (IEF), and nucleotide sequencing of PCR products. Results. Serotype 61 was predominant and correlated with genomic fingerprints of RAPD and PFGE in 11 of 15 isolates. One SHV-5- producer predominant clone with a high case-fatality rate was identified. Conclusions. Molecular biology techniques are useful tools to characterize the K. pneumoniae clone isolated from patients and health care workers, suggesting potential cross-transmission. These data call for strengthening control programs to prevent dissemination of nosocomial infections in the studied hospital.


REFERENCES

  1. González-Vértiz A, Alcántar-Curiel D, Cuauhtli M, Daza C, Gayosso C, Solache G et al. Multiresistant extended-spectrum β-lactamaseproducing Klebsiella pneumoniae causing an outbreak of nosocomial bloodstream infection. Infect Control Hosp Epidemiol 2001; 22:723-725.

  2. Vernon MO, Trick WE, Welbel SF, Peterson BJ, Weinstein RA. Adherence with hand hygiene: Does number of sinks matter? Infect Control Hosp Epidemiol 2003; 24:224-225.

  3. Jacoby G, Busch K. Aminoacid sequences for TEM, SHV and OXA extended spectrum and inhibitor resistant β-lactamasa. Lahey Clinic. Disponible en: http://www.lahey.org/Studies/?D=http://www.lahey.org/studies/webt.stm&C=404, last modified - 09/03/04.

  4. Martínez G, Alpuche C, Anaya C, Alcántara D, Gayosso C, Daza C et al. Outbreak of nosocomial sepsis and pneumonia in a Newborn

  5. Intensive Care Unit by multiresistant extended-spectrum β-Lactamaseproducing

  6. Klebsiella pneumoniae: High impact on mortality. Infect Control Hosp Epidemiol 2001; 22:725-728.

  7. Silva J, Gatica R, Aguilar C, Becerra Z, Garza U, Velázquez M et al. Outbreak of infection with extended-spectrum-β-lactamase-producing Klebsiella pneumoniae in a Mexican hospital. J Clin Microbiol 2001; 39:3193–3196.

  8. Miranda G, Castro N, Leaños B, Valenzuela A, Garza-Ramos U, Rojas T et al. Clonal and horizontal dissemination of Klebsiella pneumoniae expressing SHV-5 extended-spectrum β-Lactamase in a Mexican pediatric hospital. J Clin Microbiol 2004; 42:30-35.

  9. National Committee for Clinical Laboratory Standards. Methods for dilution antimicrobial susceptibility test for bacteria that grow aerobically. Approved standard M100-S12. Wayne (PA): National Committee for Clinical Laboratory Standards; 2002; 22:42-46.

  10. Kieser T. Factors affecting the isolation of CCC DNA from Streptomyces lividans and Escherichia coli. Plasmid 1984; 12:10-36.

  11. Miller J. Experiments in molecular genetics. Cold Spring Harbour (NY): Cold Spring Harbour Laboratory; 1992: 82-85.

  12. Edwards PR, Fife MA. Capsule types of Klebsiella. 1952; 91:92-104.

  13. Eisen D, Russell EG, Tymms M, Roper EJ, Grayson ML, Turnidge J. Random amplified polymorphic DNA and plasmid analysis used in investigation of an outbreak of multiresistant Klebsiella pneumoniae. J Clin Microbiol 1995; 33:713-717.

  14. Matthew G, Hedges R, Smith J. Types of β-lactamases determined by plasmid in gram-negative bacteria. J Bacteriol 1979; 138:657-662.

  15. Silva J, Aguilar C. ß-Lactamase bioassay: A simplified method to determine extended-spectrum ß-lactamases (ESBL) in enterobacteria. Arch Med Res 1997; 28:285-287.

  16. Kaufmann ME. Pulsed-field gel electrophoresis. Methods Mol Med 1998; 15:17-31.

  17. Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH et al. Interpreting chromosomal DNA restriction patterns produced by pulsed- field gel electrophoresis: Criteria for bacterial strain typing. J Clin Microbiol 1995; 33:2233-2239.

  18. Barthelemy M, Peduzzi J, Labia R. Complete amino acid sequence of p453-plasmid-mediated PIT-2 β-lactamase (SHV-1). J Biochem 1988; 251:73-79.

  19. Mondino SS, Castro AC, Mondino PJ, Carvalho Mda G, Silva KM, Teixeira LM. Phenotypic and genotypic characterization of clinical and intestinal enterococci isolated from inpatients and outpatient: Two Brazilian hospitals. Microb Drug Resist 2003; 9:167-174.

  20. Girou E, Azar J, Wolkenstein P, Cizeau F, Brun-Buisson C, Roujeau JC. Comparison of systematic versus selective screening for methicillinresistant Staphylococcus aureus carriage in a high-risk dermatology ward. Infect Control Hosp Epidemiol 2000; 21:583-587.

  21. Martínez JI, Baquero F. Interactions among strategies associated with bacterial infection: Pathogenicity, epidemicity, and antibiotic resistance. Clin Microbiol Rev 2002; 15:647-679.

  22. Webber M, Piddock L. The importance of efflux pumps in bacterial antibiotic resistance. J Antimicrob Chemother 2003; 51:9-11.

  23. Hernández RI, Gaitán MJ, Gaitán GE. Extrinsic contamination of intravenous infusates administered to hospitalized children in Mexico. J Pediatr Infect Dis 2000; 19: 888-890.

  24. Macías AE, Bruckner DA, Hindler JA. Parenteral infusions as culture media from a viewpoint of nosocomial bacteremia. Rev Invest Clin 2000; 52: 39-43.

  25. González-Vértiz A, Alcántar-Curiel D, Cuauhtli M, Daza C, Gayosso C, Solache G et al. Multiresistant extended-spectrum β-lactamaseproducing Klebsiella pneumoniae causing an outbreak of nosocomial bloodstream infection. Infect Control Hosp Epidemiol 2001; 22:723-725.

  26. Vernon MO, Trick WE, Welbel SF, Peterson BJ, Weinstein RA. Adherence with hand hygiene: Does number of sinks matter? Infect Control Hosp Epidemiol 2003; 24:224-225.

  27. Jacoby G, Busch K. Aminoacid sequences for TEM, SHV and OXA extended spectrum and inhibitor resistant β-lactamasa. Lahey Clinic. Disponible en: http://www.lahey.org/Studies/?D=http://www.lahey.org/studies/webt.stm&C=404, last modified-09/03/04.

  28. Martínez G, Alpuche C, Anaya C, Alcántara D, Gayosso C, Daza C et al. Outbreak of nosocomial sepsis and pneumonia in a Newborn Intensive Care Unit by multiresistant extended-spectrum β-Lactamaseproducing Klebsiella pneumoniae: High impact on mortality. Infect Control Hosp Epidemiol 2001; 22:725-728.

  29. Silva J, Gatica R, Aguilar C, Becerra Z, Garza U, Velázquez M et al. Outbreak of infection with extended-spectrum-β-lactamase-producing Klebsiella pneumoniae in a Mexican hospital. J Clin Microbiol 2001; 39:3193–3196.

  30. Miranda G, Castro N, Leaños B, Valenzuela A, Garza-Ramos U, Rojas T et al. Clonal and horizontal dissemination of Klebsiella pneumoniae expressing SHV-5 extended-spectrum β-Lactamase in a Mexican pediatric hospital. J Clin Microbiol 2004; 42:30-35.

  31. National Committee for Clinical Laboratory Standards. Methods for dilution antimicrobial susceptibility test for bacteria that grow aerobically. Approved standard M100-S12. Wayne (PA): National Committee for Clinical Laboratory Standards; 2002; 22:42-46.

  32. Kieser T. Factors affecting the isolation of CCC DNA from Streptomyces lividans and Escherichia coli. Plasmid 1984; 12:10-36.

  33. Miller J. Experiments in molecular genetics. Cold Spring Harbour (NY): Cold Spring Harbour Laboratory; 1992: 82-85.

  34. Edwards PR, Fife MA. Capsule types of Klebsiella. 1952; 91:92-104.

  35. Eisen D, Russell EG, Tymms M, Roper EJ, Grayson ML, Turnidge J. Random amplified polymorphic DNA and plasmid analysis used in investigation of an outbreak of multiresistant Klebsiella pneumoniae. J Clin Microbiol 1995; 33:713-717.

  36. Matthew G, Hedges R, Smith J. Types of β-lactamases determined by plasmid in gram-negative bacteria. J Bacteriol 1979; 138:657-662.

  37. Silva J, Aguilar C. ß-Lactamase bioassay: A simplified method to determine extended-spectrum ß-lactamases (ESBL) in enterobacteria. Arch Med Res 1997; 28:285-287.

  38. Kaufmann ME. Pulsed-field gel electrophoresis. Methods Mol Med 1998; 15:17-31.

  39. Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH et al. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: Criteria for bacterial strain typing. J Clin Microbiol 1995; 33:2233-2239.

  40. Barthelemy M, Peduzzi J, Labia R. Complete amino acid sequence of p453-plasmid-mediated PIT-2 β-lactamase (SHV-1). J Biochem 1988; 251:73-79.

  41. Mondino SS, Castro AC, Mondino PJ, Carvalho Mda G, Silva KM, Teixeira LM. Phenotypic and genotypic characterization of clinical and intestinal enterococci isolated from inpatients and outpatient: Two Brazilian hospitals. Microb Drug Resist 2003; 9:167-174.

  42. Girou E, Azar J, Wolkenstein P, Cizeau F, Brun-Buisson C, Roujeau JC. Comparison of systematic versus selective screening for methicillinresistant Staphylococcus aureus carriage in a high-risk dermatology ward. Infect Control Hosp Epidemiol 2000; 21:583-587.

  43. Martínez JI, Baquero F. Interactions among strategies associated with bacterial infection: Pathogenicity, epidemicity, and antibiotic resistance. Clin Microbiol Rev 2002; 15:647-679.

  44. Webber M, Piddock L. The importance of efflux pumps in bacterial antibiotic resistance. J Antimicrob Chemother 2003; 51:9-11.

  45. Hernández RI, Gaitán MJ, Gaitán GE. Extrinsic contamination of intravenous infusates administered to hospitalized children in Mexico. J Pediatr Infect Dis 2000; 19: 888-890.

  46. Macías AE, Bruckner DA, Hindler JA. Parenteral infusions as culture media from a viewpoint of nosocomial bacteremia. Rev Invest Clin 2000; 52: 39-43.




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