StaphVar-DNA microarray analysis of accessory genome elements of community-acquired methicillin-resistant Staphylococcus aureus.
OBJECTIVE
Approximately 75% of the genome of Staphylococcus aureus (the 'core' genome) is highly conserved between strains, whereas the remaining 25% (the 'accessory' genome) is composed of mobile genetic elements (MGEs), containing virulence and resistance genes. We developed a composite microarray focused on resistance and virulence genes located on the accessory or core-variable genome to characterize a collection of Belgian community-acquired methicillin-resistant S. aureus (CA-MRSA) strains.


METHODS
Oligonucleotide probes targeting 403 genes encoding antimicrobial resistance (35%), virulence (28%) and adhesion (31%) factors were designed among eight S. aureus sequenced genomes. The StaphVar Array was validated by testing five of the strains used for the design and utilized to characterize 13 CA-MRSA strains representative of the multilocus sequence typing (MLST) sequence types circulating in Belgium.


RESULTS
Analysis of the gene content of the five reference strains by the StaphVar Array matched 90% to 97% of the theoretical results. Analysis of CA-MRSA strains showed that 54.4% of the genes tested were strain-dependent. Strains presented specific exotoxin, enterotoxin, cytolysin and adhesin gene profiles by MLST lineage. One exception to these 'lineage-specific' profiles was the variable presence of the arginine catabolic mobile element (characteristic of the USA300 clone) within ST8 strains.


CONCLUSIONS
The StaphVar Array enables the characterization of approximately 400 variable resistance and virulence determinants in S. aureus. CA-MRSA strains displayed extensive diversity in virulence and resistance profiles. The presence of the USA300 clone in Belgium was confirmed. Although mainly located on MGEs, associations of virulence genes were highly conserved within strains of the same MLST lineage.
El Garch
F
F
Université Libre de Bruxelles, Hôpital Erasme, Belgium.
Hallin
M
M
De Mendonça
R
R
Denis
O
O
Lefort
A
A
Struelens
M J
MJ
D018829:Adhesins, Bacterial / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000379:methods*; D001530:Belgium; D017714:Community-Acquired Infections / Q000382:microbiology*; D024881:Drug Resistance, Bacterial; D005798:Genes, Bacterial; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D046228:Microarray Analysis / Q000379:methods*; D011110:Polymorphism, Genetic; D013203:Staphylococcal Infections / Q000382:microbiology*; D037521:Virulence Factors / Q000235:genetics
D018829:Adhesins, Bacterial; D037521:Virulence Factors