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Journal of Clinical Microbiology, June 2007, p. 1830-1837, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.02402-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of Molecular Typing Methods in Characterizing a European Collection of Epidemic Methicillin-Resistant Staphylococcus aureus Strains: the HARMONY Collection{triangledown}

Barry D. Cookson,1 D. Ashley Robinson,2 Alastair B. Monk,3 Stephen Murchan,4 Ariane Deplano,5 Rafaël de Ryck,5 Marc J. Struelens,5 Christina Scheel,6 Vivian Fussing,6 Saara Salmenlinna,7 Jaana Vuopio-Varkila,7 Christina Cuny,8 Wolfgang Witte,8 Panayotis T. Tassios,9 Nikolas J. Legakis,9 Willem van Leeuwen,10 Alex van Belkum,10 Anna Vindel,11 Javier Garaizar,12 Sara Haeggman,13 Barbro Olsson-Liljequist,13 Ulrika Ransjo,14 Manica Muller-Premru,15 Waleria Hryniewicz,16 Angela Rossney,17 Brian O'Connell,17 Benjamin D. Short,18 Jonathan Thomas,18 Simon O'Hanlon,18 and Mark C. Enright18*

Laboratory of Hospital Infection, Centre for Infections, Health Protection Agency, London, United Kingdom,1 New York Medical School, New York, New York,2 Virginia Commonwealth University, Richmond, Virginia,3 Health Protection Surveillance Centre, Dublin, Ireland,4 Laboratoire de Microbiologie, Hopital Erasme, Bruxelles, Belgium,5 Statens Serum Institut, Copenhagen, Denmark,6 National Public Health Laboratory (KTL), Helsinki, Finland,7 Robert Koch-Institut, Wernigerode (Harz), Germany,8 Department of Microbiology, University of Athens, Athens, Greece,9 Erasmus MC Center, Rotterdam, The Netherlands,10 Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain,11 Dpt. Immunol., Microbiol. y Parasitol., F. Farmacia, UPV/EHU, Vitoria-Gasteiz, Spain,12 Swedish Institute for Infectious Disease Control, Solna, Sweden,13 Swedish Institute for Infectious Disease Control, Solna, Sweden,14 Institute of Microbiology, University of Ljubljana, Ljubljana, Slovenia,15 Sera and Vaccines Central Research Laboratory, Warsaw, Poland,16 National MRSA Reference Laboratory, St James's Hospital, James's St., Dublin 8, Ireland,17 Department of Infectious Disease Epidemiology, Imperial College, London, United Kingdom,18

Received 29 November 2006/ Returned for modification 8 January 2007/ Accepted 1 March 2007

We analyzed a representative sample of methicillin-resistant Staphylococcus aureus (MRSA) from 11 European countries (referred to as the HARMONY collection) using three molecular typing methods used within the HARMONY group to examine their usefulness for large, multicenter MRSA surveillance networks that use these different laboratory methodologies. MRSA isolates were collected based on their prevalence in each center and their genetic diversity, assessed by pulsed-field gel electrophoresis (PFGE). PFGE groupings (≤3 bands difference between patterns) were compared to those made by sequencing of the variable repeats in the protein A gene spa and clonal designations based on multilocus sequence typing (MLST), combined with PCR analysis of the staphylococcal chromosome cassette containing the mec genes involved in methicillin resistance (SCCmec). A high level of discrimination was achieved using each of the three methodologies, with discriminatory indices between 89.5% and 91.9% with overlapping 95% confidence intervals. There was also a high level of concordance of groupings made using each method. MLST/SCCmec typing distinguished 10 groups containing at least two isolates, and these correspond to the majority of nosocomial MRSA clones described in the literature. PFGE and spa typing resolved 34 and 31 subtypes, respectively, within these 10 MRSA clones, with each subtype differing only slightly from the most common pattern using each method. The HARMONY group has found that the methods used in this study differ in their availability and affordability to European centers involved in MRSA surveillance. Here, we demonstrate that the integration of such technologies is achievable, although common protocols (such as we have developed for PFGE) may also be important, as is the use of centralized Internet sites to facilitate data analysis. PFGE and spa-typing data from analysis of MRSA isolates from the many centers that have access to the relevant equipment can be compared to reference patterns/sequences, and clonal designations can be made. In the majority of cases, these will correspond to those made by the (more expensive) method of choice—MLST/SCCmec typing—and these alternative methods can therefore be used as frontline typing systems for multicenter surveillance of MRSA.

* Corresponding author. Mailing address: Department of Infectious Disease Epidemiology, Imperial College, London, United Kingdom. Phone: 44 207 5943417. Fax: 44 207 594 3693. E-mail: m.c.enright{at}imperial.ac.uk

{triangledown} Published ahead of print on 11 April 2007.

Journal of Clinical Microbiology, June 2007, p. 1830-1837, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.02402-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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