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Antimicrobial Agents and Chemotherapy, January 2005, p. 366-379, Vol. 49, No. 1
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.1.366-379.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Analysis of the Genetic Variability of Virulence-Related Loci in Epidemic Clones of Methicillin-Resistant Staphylococcus aureus

A. R. Gomes,1 S. Vinga,2 M. Zavolan,3 and H. de Lencastre1,4*

Laboratory of Molecular Genetics,1 Biomathematics Group, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal,2 Laboratory of Computational Genomics,3 Laboratory of Microbiology, The Rockefeller University, New York, New York4

Received 26 November 2003/ Returned for modification 6 February 2004/ Accepted 11 September 2004

Methicillin-resistant Staphylococcus aureus (MRSA) isolates have previously been classified into major epidemic clonal types by pulsed-field gel electrophoresis in combination with multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec typing. We aimed to investigate whether genetic variability in potentially polymorphic domains of virulence-related factors could provide another level of differentiation in a diverse collection of epidemic MRSA clones. The target regions of strains representative of epidemic clones and genetically related methicillin-susceptible S. aureus isolates from the 1960s that were sequenced included the R domains of clfA and clfB; the D, W, and M regions of fnbA and fnbB; and three regions in the agr operon. Sequence variation ranged from very conserved regions, such as those for RNAIII and the agr interpromoter region, to the highly polymorphic R regions of the clf genes. The sequences of the clf R domains could be grouped into six major sequence types on the basis of the sequences in their 3' regions. Six sequence types were also observed for the fnb sequences at the amino acid level. From an evolutionary point of view, it was interesting that a small DNA stretch at the 3' clf R-domain sequence and the fnb sequences agreed with the results of MLST for this set of strains. In particular, clfB R-domain sequences, which had a high discriminatory capacity and with which the types distinguished were congruent with those obtained by other molecular typing methods, have potential for use for the typing of S. aureus. Clone- and strain-specific sequence motifs in the clf and fnb genes may represent useful additions to a typing methodology with a DNA array.

* Corresponding author. Mailing address: The Rockefeller University, 1230 York Ave., New York, NY 10021. Phone: (212) 327-8278. Fax: (212) 327-8688. E-mail: lencash{at}rockefeller.edu.

Antimicrobial Agents and Chemotherapy, January 2005, p. 366-379, Vol. 49, No. 1
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.1.366-379.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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