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Journal of Clinical Microbiology, October 2000, p. 3527-3533, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Multicenter Evaluation of Epidemiological Typing of Methicillin-Resistant Staphylococcus aureus Strains by Repetitive-Element PCR Analysis

Ariane Deplano,1 Annette Schuermans,2 Johan Van Eldere,2 Wolfgang Witte,3 Hèléne Meugnier,4 Jerome Etienne,4 Hajo Grundmann,5 Daniel Jonas,5 Gerda T. Noordhoek,6 Jolanda Dijkstra,6 Alex van Belkum,7 Willem van Leeuwen,7 Panayotis T. Tassios,8 Nicholas J. Legakis,8 Anneke van der Zee,9 Anneke Bergmans,9 Dominique S. Blanc,10 Fred C. Tenover,11 Barry C. Cookson,12 Gael O'Neil,12 Marc J. Struelens,1,* and The European Study Group on Epidemiological Markers of the ESCMID

Reference Laboratory for Staphylococci, Université Libre de Bruxelles, Hôpital Erasme, Department of Microbiology, 1070 Brussels, Belgium1; Rega Institute for Medical Research, Department of Microbiology & Immunology, B-3000 Leuven, Belgium2; Robert Koch Institute BGA, D-38855 Wernigerode, Germany3; Hôpital Edouard Herriot, Laboratoire Central de Microbiologie, 69437 Lyon cedex 03, France4; Klinikum der Albert-Ludwigsuniversitat, Institut fur Umweltmedizin und Krankenhaushygiene, D-79106 Freiburg, Germany5; Public Health Laboratory Friesland, 8900 JA Leeuwarden, The Netherlands6; University Hospital Rotterdam, Department of Bacteriology, 3015 GD Rotterdam, The Netherlands7; Department of Microbiology, Medical School, University of Athens, 115 27 Athens, Greece8; Laboratory of Molecular Microbiology, St. Elisabeth Hospital Tilburg, 5000 AS Tilburg, The Netherlands9; Centre Hospitalier Universitaire Vaudois, CH 1011 Lausanne, Switzerland10; Nosocomial Pathogens Laboratory Branch, Hospital Infections Program G-08, Centers for Disease Control and Prevention, Atlanta, Georgia 3033311; and Hospital Infection Laboratory, Central Public Health Laboratory, London NW95HT, United Kingdom12

Received 12 February 2000/Returned for modification 3 May 2000/Accepted 30 June 2000

Rapid and efficient epidemiologic typing systems would be useful to monitor transmission of methicillin-resistant Staphylococcus aureus (MRSA) at both local and interregional levels. To evaluate the intralaboratory performance and interlaboratory reproducibility of three recently developed repeat-element PCR (rep-PCR) methods for the typing of MRSA, 50 MRSA strains characterized by pulsed-field gel electrophoresis (PFGE) (SmaI) analysis and epidemiological data were blindly typed by inter-IS256, 16S-23S ribosomal DNA (rDNA), and MP3 PCR in 12 laboratories in eight countries using standard reagents and protocols. Performance of typing was defined by reproducibility (R), discriminatory power (D), and agreement with PFGE analysis. Interlaboratory reproducibility of pattern and type classification was assessed visually and using gel analysis software. Each typing method showed a different performance level in each center. In the center performing best with each method, inter-IS256 PCR typing achieved R = 100% and D = 100%; 16S-23S rDNA PCR, R = 100% and D = 82%; and MP3 PCR, R = 80% and D = 83%. Concordance between rep-PCR type and PFGE type ranged by center: 70 to 90% for inter-IS256 PCR, 44 to 57% for 16S-23S rDNA PCR, and 53 to 54% for MP3 PCR analysis. In conclusion, the performance of inter-IS256 PCR typing was similar to that of PFGE analysis in some but not all centers, whereas other rep-PCR protocols showed lower discrimination and intralaboratory reproducibility. None of these assays, however, was sufficiently reproducible for interlaboratory exchange of data.

* Corresponding author. Mailing address: Service de Microbiologie, Université Libre de Bruxelles, Hôpital Erasme, Route de Lennik, 808. 1070 Brussels, Belgium. Phone: 32.2.555 45 18. Fax: 32 2 555 64 59. E-mail: marc.struelens{at}ulb.ac.be.

Journal of Clinical Microbiology, October 2000, p. 3527-3533, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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