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Molecular Markers for Differentiation of Multiresistant Klebsiella Pneumoniae Isolates in a Pediatric Hospital

Published online by Cambridge University Press:  02 January 2015

Sylvie Lhopital
Affiliation:
Service of Microbrblogy, Intensive-Care Surgery, and Gastroenterology, Hôpital Robert Debré, Paris, France
Stephane Bonacorsi
Affiliation:
Service of Microbrblogy, Intensive-Care Surgery, and Gastroenterology, Hôpital Robert Debré, Paris, France
Daniele Meis
Affiliation:
Service of Microbrblogy, Intensive-Care Surgery, and Gastroenterology, Hôpital Robert Debré, Paris, France
Naima Brahimi
Affiliation:
Service of Microbrblogy, Intensive-Care Surgery, and Gastroenterology, Hôpital Robert Debré, Paris, France
Stephanie Mathy
Affiliation:
Service of Microbrblogy, Intensive-Care Surgery, and Gastroenterology, Hôpital Robert Debré, Paris, France
Jean Navarro
Affiliation:
Service of Microbrblogy, Intensive-Care Surgery, and Gastroenterology, Hôpital Robert Debré, Paris, France
Yves Aigrain
Affiliation:
Service of Microbrblogy, Intensive-Care Surgery, and Gastroenterology, Hôpital Robert Debré, Paris, France

Abstract

Objective:

To study the spread of extended spectrum ß-lactamase-producing, but aminoglycoside-susceptible, Klebsiella pneumoniae strains in our hospital over an 8-month period, by using two genotypic markers.

Design:

Ribotyping (using two endonucleases) and randomly amplified polymorphic DNA analysis (RAPD; using two different 10-mer primers) were applied to the epidemiological typing of clinical K pneumoniae isolates from stools, ileal fluid, or urine of hospitalized children.

Setting and Patients:

The surgical intensive care ward (S1: 9 patients, 17 isolates), surgical unit (S2: 2 patients, 2 isolates), and gastroenterology ward (GE: 1 patient, 1 isolate) of the Robert Debre Hospital of Paris, France.

Results:

Ribotyping of the 20 clinical isolates, the type strain of the species, and two epidemiologically unrelated isolates with EcoRI and HindlII revealed 6 and 5 different patterns, respectively. Six ribotypes were identified by using these two enzymes. RAPD generated 6 distinct patterns, in complete agreement with ribotyping. Our geno-typic results showed that 11 patients from wards Sl, S2, and GE harbored genotypically related strains, suggesting nosocomial transmission and cross-colonization between and within the three wards.

Conclusions:

Ribotyping and RAPD appear to be reliable methods for distinguishing K pneumoniae strains. The spread of one strain of K pneumoniae in different units of our hospital was demonstrated by both methods. However, RAPD has the advantage of simplicity and rapidity conferred by polymerase chain reaction.

Type
Original Articles
Copyright
Copyright © The Society for Healthcare Epidemiology of America 1997

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References

1. Jarlier, V, Nicolas, MH, Fournier, G, Philippon A Extended broad-spectrum ß-lactamases conferring transferable resistance to newer ß-lactam agents in Enterobacteriacae: hospital prevalence and susceptibility patterns. Reviews of Infectious Diseases 1988;10:867878.Google Scholar
2. Mulgrave, L. Extended broad-spectrum ß-lactamases in Australia. Med J Aust 1990;152:444445.Google Scholar
3. Rice, L, Willey, S, Papanicolaou, G, et al. Outbreak of ceftazidime resistance caused by extended-spectrum ß-lactamases at a Massachusetts chronic-care facility. Antimicrob Agents Chemother 1990;34:21932199.Google Scholar
4. Legrand, P, Former, G, Bure, A, et al. Detection of extended broad-spectrum ß-lactamase in Enterobacteriaceae in four French hospitals. Eur J Clin Microbiol Infect Dis 1989;8:527529.CrossRefGoogle Scholar
5. Philippon, A, Labia, R, Jacoby, G. Extended-Spectrum ß-lactamases. Antiomicrob Agents Chemother 1989;33:11311136.Google Scholar
6. Arlet, G, Sanson-Le Pors, M, Rouveau, M, et al. Outbreak of nosocomial infections due to Klebsiella pneumoniae producing SHV-4 ß-lactamase. Eur J Clin Microbial Infect Dis 1990;9:797803.Google Scholar
7. National Committee for Clinical Laboratory Standards. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically. Approved standard M7-A2 (M100-S4). Villanova, PA: NCCLS; 1993.Google Scholar
8. Bingen, E, Denamur, E, Lambert-Zechovsky, N, et al. DNA restriction fragment-length polymorphism differentiates crossed from independent infections in nosocomial Xanthomonas maltophilia bacteremia. J Clin Microbiol 1991;29:13481350.Google Scholar
9. Williams, J, Kubelik, A, Livak, K, Rafalski, J, Tingey, S. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acid Res 1990;18:65316535.Google Scholar
10. Bingen, E, Boissinot, C, Desjardins, P, et al. Arbitrarily primed polymerase chain reaction provides rapid differentiation of Proteus mirabilis isolates from a pediatric hospital. J Clin Microbiol 1993;31:10551059.CrossRefGoogle ScholarPubMed
11. Bingen, E, Mariani-Kurkdjian, P, Lambert-Zechovsky, N, et al. Ribotyping provides efficient differentiation of nosocomial Serratia marcese ens isolates in a pediatric hospital. J Clin Microbial 1992;30:20882091.Google Scholar
12. Kliebe, C, Nies, B, Meyer, J, Tolxdorff-Neutzling, , Wiedemann, B. Evolution of plasmid-coded resistance to broad-spectrum cephalosporins. Antimicrobiol Agents Chemother 1985;28:302307.CrossRefGoogle ScholarPubMed
13. Fernandez-Rodriguez, A, Canton, R, Perez-Diaz, JC, Martinez-Beltran, J, Picazo, JJ, Baquero, F. Aminoglycoside-modifying enzymes in clinicai isolates harboring extended-spectrum ß-lactamases. Antimicrob Agents Chemother 1992;11:25362538.Google Scholar
14. Lambert-Zechovsky, N, Bergen, E, Denamur, E, et al. Molecular analysis provides evidence for the endogenous origin of bacteremia and meningitis due to Enterobacter cloacaae in an infant. Clin Inect Dis 1992;15:3032.Google Scholar
15. Orskov, I, Orskov, F. Serotyping of Klebsiella . Methods in Microbiology 1984;14:143164.Google Scholar
16. Rubin, S. Klebsiella markers system. Infect Control 1985;6:5963.Google Scholar
17. Bingen, E. Applications of molecular methods to epidemiologic investigations of nosocomial infections in a pediatric hospital. Infect Control Hosp Epidemiol 1994;15:488493.Google Scholar
18. Mayer, L. Use of plasmiti profiles in epidemiologic surveillance of disease outbreaks and in tracing the transmission of antibiotic resistance. Clin Microbiol Rev 1988;1:228243.Google Scholar
19. Bingen, E, Desjardins, P, Arlet, G, et al. Molecular epidemiology of plasmid spread among extended broad-spectrum ß-lactamase-producing Klebsiella pneumoniae isolates in a pediatric hospital. J Clin Microbial 1993;31:179184.Google Scholar
20. Cookson, B, Johnson, A, Azadian, B, et al. International inter- and intrahospital patient spread of a multiple antibiotic-resistant strain of Klebsiella pneumoniae . J Infect Dis 1995;171:511513.Google Scholar
21. Tenover, F, Arbeit, R, Goering, V, et al. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol 1995;33:22332239.Google Scholar
22. Grimont, F, Grimont, P. Ribosomal ribonucleic acid gene restriction patterns as potential taxonomic tools. Annales de l'Institut Pasteur –; Microbiologie (Paris) 1986;137B:165175.Google Scholar
23. Stull, T, Lipuma, J, Edlind, T. A broad spectrum probe for molecular epidemiology of bacteria: ribosomal RNA J Infect Dis 1988;157:280286.CrossRefGoogle ScholarPubMed
24. Bingen, E, Denamur, E, Elion, J. Use of ribotyping in epidemiological surveillance of nosocomial outbreaks. Clin Microbiol 1994;7:311327.Google Scholar
25. Welsh, J, McClelland, M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 1990:18:72137218.Google Scholar
26. Eisen, D, Russell, E, Tymms, M, Roper, E, Grayson, M, Turnidge, J. Random amplified polymorphic DNA and plasmid analyses used in investigation of an outbreak of multiresistant Klemetta pneumoniae . J Clin Microbiol 1995;33:713717.Google Scholar
27. Kerr, J, Moore, J, Curran, M, et al. Investigation of a nosocomial outbreak of Pseudomonas aeruginosa pneumonia in an intensive care unit by random amplification of polymorphic DNA assay. J Hosp Infect 1995;30:125131.Google Scholar
28. Cave, H, Bingen, E, Elion, J, Denamur, E. Differentiation of Escherichia coli strains using randomly amplified polymorphic DNA analysis. Res Microbiol 1994;145:141150.Google Scholar
29. Brun-Buisson, C, Legrand, P, Philippon, A, Montravers, F, Ansquer, M, Duval, J. Transferable enzymatic resistance to third-generation cephalosporins during nosocomial outbreak of multiresistant Klebsiella pneumoniae . Lancet 1987;ii:302306.Google Scholar