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Journal of Clinical Microbiology, June 2009, p. 1620-1627, Vol. 47, No. 6
0095-1137/09/$08.00+0 doi:10.1128/JCM.01579-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Methicillin-Resistant Staphylococcus aureus in Spain: Molecular Epidemiology and Utility of Different Typing Methods 
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Ana Vindel,1
Oscar Cuevas,2
Emilia Cercenado,2*
Carmen Marcos,1
Verónica Bautista,1
Carol Castellares,2
Pilar Trincado,1
Teresa Boquete,1
Maria Pérez-Vázquez,1
Mercedes Marín,2
Emilio Bouza,2 and the Spanish Group for the Study of Staphylococcus
Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain,1 Servicio de Microbiología, Hospital General Universitario Gregorio Marañón, Madrid, Spain2
Received 14 August 2008/ Returned for modification 12 November 2008/ Accepted 21 March 2009
In a point-prevalence study performed in 145 Spanish hospitals in 2006, we collected 463 isolates of Staphylococcus aureus in a single day. Of these, 135 (29.2%) were methicillin (meticillin)-resistant S. aureus (MRSA) isolates. Susceptibility testing was performed by a microdilution method, and mecA was detected by PCR. The isolates were analyzed by pulsed-field gel electrophoresis (PFGE) after SmaI digestion, staphylococcal chromosomal cassette mec (SCCmec) typing, agr typing, spa typing with BURP (based-upon-repeat-pattern) analysis, and multilocus sequence typing (MLST). The 135 MRSA isolates showed resistance to ciprofloxacin (93.3%), tobramycin (72.6%), gentamicin (20.0%), erythromycin (66.7%), and clindamycin (39.3%). Among the isolates resistant to erythromycin, 27.4% showed the M phenotype. All of the isolates were susceptible to glycopeptides. Twelve resistance patterns were found, of which four accounted for 65% of the isolates. PFGE revealed 36 different patterns, with 13 major clones (including 2 predominant clones with various antibiotypes that accounted for 52.5% of the MRSA isolates) and 23 sporadic profiles. Two genotypes were observed for the first time in Spain. SCCmec type IV accounted for 6.7% of the isolates (70.1% were type IVa, 23.9% were type IVc, 0.9% were type IVd, and 5.1% were type IVh), and SCCmec type I and SCCmec type II accounted for 7.4% and 5.2% of the isolates, respectively. One isolate was nontypeable. Only one of the isolates produced the Panton-Valentine leukocidin. The isolates presented agr type 2 (82.2%), type 1 (14.8%), and type 3 (3.0%). spa typing revealed 32 different types, the predominant ones being t067 (48.9%) and t002 (14.8%), as well as clonal complex 067 (78%) by BURP analysis. The MRSA clone of sequence type 125 and SCCmec type IV was the most prevalent throughout Spain. In our experience, PFGE, spa typing, SCCmec typing, and MLST presented good correlations for the majority of the MRSA strains; we suggest the use of spa typing and PFGE typing for epidemiological surveillance, since this combination is useful for both long-term and short-term studies.
* Corresponding author. Mailing address: Servicio de Microbiología, Hospital General Universitario Gregorio Marañón, C/ Dr. Esquerdo, 46, Madrid 28007, Spain. Phone: 34-91-586-8459. Fax: 34-91-504-4906. E-mail: ecercenado{at}terra.es
Published ahead of print on 1 April 2009.
Supplemental material for this article may be found at http://jcm.asm.org/.
Journal of Clinical Microbiology, June 2009, p. 1620-1627, Vol. 47, No. 6
0095-1137/09/$08.00+0 doi:10.1128/JCM.01579-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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