Analysis on distribution and genomic diversity of high-level antiseptic resistance genes qacA and qacB in human clinical isolates of Staphylococcus aureus.
High-level antiseptic resistance of Staphylococcus aureus is mediated by multidrug efflux pumps encoded by qacA and qacB genes. We investigated distribution and genomic diversity of these antiseptic resistance genes in a total of 522 clinical strains of S. aureus isolated recently in a Japanese hospital. The qacA/B gene was detected in 32.6% of methicillin-resistant S. aureus (MRSA) and 7.5% of methicillin-susceptible S. aureus (MSSA), whereas the low-level resistance gene smr, which was examined simultaneously, was detected at lower frequencies in both MRSA (3.3%) and MSSA (5.9%). Epidemiologic typing of S. aureus isolates suggested that higher prevalence of qacA/B in MRSA may be due to spread of a single predominant MRSA strain carrying qacA/B in the hospital. Restriction fragment length polymorphism (RFLP) analysis indicated higher prevalence of the qacB-type gene (59.3%) than the qacA-type gene (40.7%) among the qacA/B genes detected. Nucleotide sequencing analysis revealed the presence of two genetic variants in qacA (V1 and V2) and four variants in qacB (V1-V4) that differ from the qacA prototype in pSK1 by 1-5 nucleotides and 7-9 nucleotides, respectively. Although most strains with qacA-V1, qacA-V2, qacB-V3, and qacB-V4 showed high-level resistance to ethidium bromide (EB)(MIC > 100 microg/ml), all of the S. aureus isolates carrying qacB-V1 and qacB-V2 showed lower MICs of EB and some monovalent cationic antiseptic substances. By analysis of the genomic organization of the qacA/B downstream region, divergent forms of this region rearranged with an insertion of IS256 or IS257 were found primarily for qacB. The downstream region of qacA-V1 was suggested to be an evolutionary origin for other divergent forms. These findings indicated that both qacA and qacB are prevalent in recent clinical isolates, especially in MRSA, and these genes consist of variable genetic variants that may be responsible for different resistance levels against antiseptic substances.
Alam
Mohammed Mahbub
MM
Department of Hygiene, Sapporo Medical University School of Medicine, Sapporo 060-8556, Japan.
Kobayashi
Nobumichi
N
Uehara
Nobuyuki
N
Watanabe
Naoki
N
D000891:Anti-Infective Agents, Local / Q000378:metabolism / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics*; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics*; D016106:Methicillin Resistance / Q000235:genetics; D008969:Molecular Sequence Data; D012150:Polymorphism, Restriction Fragment Length; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013203:Staphylococcal Infections / Q000453:epidemiology / Q000382:microbiology; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism
D000891:Anti-Infective Agents, Local; D001426:Bacterial Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial; D026901:Membrane Transport Proteins; C099272:QacB protein, Staphylococcus aureus; C068498:qacA protein, Staphylococcus aureus