title: Analysis of the genetic variability of virulence-related loci in epidemic clones of methicillin-resistant Staphylococcus aureus.
abstract: 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.
authors.affiliation: Laboratory of Molecular Genetics, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: A R
authors.initials: AR
authors.lastname: Gomes
authors.forename: S
authors.initials: S
authors.lastname: Vinga
authors.forename: M
authors.initials: M
authors.lastname: Zavolan
authors.forename: H
authors.initials: H
authors.lastname: de Lencastre
mesh_terms: D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D004196:Disease Outbreaks; D016521:Electrophoresis, Gel, Pulsed-Field; D014644:Genetic Variation; D006801:Humans; D016106:Methicillin Resistance / Q000235:genetics*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013203:Staphylococcal Infections / Q000453:epidemiology* / Q000382:microbiology; D013211:Staphylococcus aureus / Q000145:classification / Q000187:drug effects / Q000235:genetics / Q000472:pathogenicity*; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics; D014774:Virulence
chemical_list: D018829:Adhesins, Bacterial; C095698:Agr protein, Staphylococcus aureus; D001426:Bacterial Proteins; D015534:Trans-Activators; C078635:fibronectin-binding proteins, bacterial