This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Seifert, H. Articles by Dijkshoorn, L. Search for Related Content PubMed PubMed Citation Articles by Seifert, H. Articles by Dijkshoorn, L.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, September 2005, p. 4328-4335, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4328-4335.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Standardization and Interlaboratory Reproducibility Assessment of Pulsed-Field Gel Electrophoresis-Generated Fingerprints of Acinetobacter baumannii

Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany,¹ Dipartimento di Scienze Biomediche, University of Trieste, Trieste, Italy,² Leiden University Medical Center, Leiden,³ National Institute for Public Health and the Environment, Bilthoven, The Netherlands⁴

Received 21 February 2005/ Returned for modification 29 April 2005/ Accepted 1 June 2005

A standard procedure for pulsed-field gel electrophoresis (PFGE) of macrorestriction fragments of Acinetobacter baumannii was set up and validated for its interlaboratory reproducibility and its potential for use in the construction of an Internet-based database for international monitoring of epidemic strains. The PFGE fingerprints of strains were generated at three different laboratories with ApaI as the restriction enzyme and by a rigorously standardized procedure. The results were analyzed at the respective laboratories and also centrally at a national reference institute. In the first phase of the study, 20 A. baumannii strains, including 3 isolates each from three well-characterized hospital outbreaks and 11 sporadic strains, were distributed blindly to the participating laboratories. The local groupings of the isolates in each participating laboratory were identical and allowed the identification of the epidemiologically related isolates as belonging to three clusters and identified all unrelated strains as distinct. Central pattern analysis by using the band-based Dice coefficient and the unweighted pair group method with mathematical averaging as the clustering algorithm showed 95% matching of the outbreak strains processed at each local laboratory and 87% matching of the corresponding strains if they were processed at different laboratories. In the second phase of the study, 30 A. baumannii isolates representing 10 hospital outbreaks from different parts of Europe (3 isolates per outbreak) were blindly distributed to the three laboratories, so that each laboratory investigated 10 epidemiologically independent outbreak isolates. Central computer-assisted cluster analysis correctly identified the isolates according to their corresponding outbreak at an 87% clustering threshold. In conclusion, the standard procedure enabled us to generate PFGE fingerprints of epidemiologically related A. baumannii strains at different locations with sufficient interlaboratory reproducibility to set up an electronic database to monitor the geographic spread of epidemic strains.

This article has been cited by other articles:

• Peleg, A. Y., Seifert, H., Paterson, D. L. (2008). Acinetobacter baumannii: Emergence of a Successful Pathogen. Clin. Microbiol. Rev. 21: 538-582 [Abstract] [Full Text]  
• Loli, A., Tzouvelekis, L. S., Gianneli, D., Tzelepi, E., Miriagou, V. (2008). Outbreak of Acinetobacter baumannii with Chromosomally Encoded VIM-1 Undetectable by Imipenem-EDTA Synergy Tests. Antimicrob. Agents Chemother. 52: 1894-1896 [Full Text]  
• Shelburne, S. A. III, Singh, K. V., White, A. C. Jr., Byrne, L., Carmer, A., Austin, C., Graviss, E., Stager, C., Murray, B. E., Atmar, R. L. (2008). Sequential Outbreaks of Infections by Distinct Acinetobacter baumannii Strains in a Public Teaching Hospital in Houston, Texas. J. Clin. Microbiol. 46: 198-205 [Abstract] [Full Text]  
• Endimiani, A., Luzzaro, F., Migliavacca, R., Mantengoli, E., Hujer, A. M., Hujer, K. M., Pagani, L., Bonomo, R. A., Rossolini, G. M., Toniolo, A. (2007). Spread in an Italian Hospital of a Clonal Acinetobacter baumannii Strain Producing the TEM-92 Extended-Spectrum {beta}-Lactamase. Antimicrob. Agents Chemother. 51: 2211-2214 [Abstract] [Full Text]  
• Peleg, A. Y., Adams, J., Paterson, D. L. (2007). Tigecycline Efflux as a Mechanism for Nonsusceptibility in Acinetobacter baumannii. Antimicrob. Agents Chemother. 51: 2065-2069 [Abstract] [Full Text]  
• Lee, J. H., Choi, C. H., Kang, H. Y., Lee, J. Y., Kim, J., Lee, Y. C., Seol, S. Y., Cho, D. T., Kim, K. W., Song, D. Y., Lee, J. C. (2007). Differences in phenotypic and genotypic traits against antimicrobial agents between Acinetobacter baumannii and Acinetobacter genomic species 13TU. J Antimicrob Chemother 59: 633-639 [Abstract] [Full Text]  
• Valenzuela, J. K., Thomas, L., Partridge, S. R., van der Reijden, T., Dijkshoorn, L., Iredell, J. (2007). Horizontal Gene Transfer in a Polyclonal Outbreak of Carbapenem-Resistant Acinetobacter baumannii. J. Clin. Microbiol. 45: 453-460 [Abstract] [Full Text]  
• Li, J., Rayner, C. R., Nation, R. L., Owen, R. J., Spelman, D., Tan, K. E., Liolios, L. (2006). Heteroresistance to Colistin in Multidrug-Resistant Acinetobacter baumannii.. Antimicrob. Agents Chemother. 50: 2946-2950 [Abstract] [Full Text]  
• Canduela, M. J., Gallego, L., Sevillano, E., Valderrey, C., Calvo, F., Perez, J. (2006). Evolution of multidrug-resistant Acinetobacter baumannii isolates obtained from elderly patients with respiratory tract infections. J Antimicrob Chemother 57: 1220-1222 [Abstract] [Full Text]