Characterization of two newly identified genes, vgaD and vatH, [corrected] conferring resistance to streptogramin A in Enterococcus faecium.

We characterized two new streptogramin A resistance genes from quinupristin-dalfopristin-resistant Enterococcus faecium JS79, which was selected from 79 E. faecium isolates lacking known genes encoding streptogramin A acetyltransferase. A 5,650-bp fragment of HindIII-digested plasmid DNA from E. faecium JS79 was cloned and sequenced. The fragment contained two open reading frames carrying resistance genes related to streptogramin A, namely, genes for an acetyltransferase and an ATP efflux pump. The first open reading frame comprised 648 bp encoding 216 amino acids with a predicted left-handed parallel ß-helix domain structure; this new gene was designated vatH. [corrected] The second open reading frame consisted of 1,575 bp encoding 525 amino acids with two predicted ATPase binding cassette transporters comprised of Walker A, Walker B, and LSSG motifs; this gene was designated vgaD. vgaD is located 65 bp upstream from vatH, [corrected] was detected together with vatH [corrected] in 12 of 179 quinupristin-dalfopristin-resistant E. faecium isolates, and was located on the same plasmid. Also, the 5.6-kb HindIII-digested fragment which was observed in JS79 was detected in nine vgaD- and vatH-containing [corrected] E. faecium isolates by Southern hybridization. Therefore, it was expected that these two genes were strongly correlated with each other and that they may be composed of a transposon. Importantly, vgaD is the first identified ABC transporter conferring resistance to streptogramin A in E. faecium. Pulsed-field gel electrophoresis patterns and sequence types of vgaD- and vatH-containing [corrected] E. faecium isolates differed for isolates from humans and nonhumans.

Restriction fragment length polymorphism (RFLP) patterns and sequence analysis of high-level mupirocin-resistant meticillin-resistant staphylococci.

High-level mupirocin resistance results from the acquisition of a mupirocin resistance (Mupr) plasmid carrying the mupA gene. In this study, we investigated the heterogeneous location of the mupA gene as well as sequence variations in mupA restriction fragment length polymorphism (RFLP) types of high-level mupirocin-resistant (MuH) staphylococci isolated from tertiary hospitals and long-term care facilities in South Korea. RFLP patterns of the mupA gene were investigated in 14 MuH staphylococci isolates, and sequence variations of the cassette-like construction composed of the transfer gene complex (trs), an insertion sequence (IS257-like) and the mupA gene of the Mupr plasmid were also studied. Among the 14 isolates, four different EcoRI/HindIII banding patterns were observed, which were determined to be caused by sequence deletion between the mupA gene and trsLM of the trs gene complex. Four different sequence types were also identified for the trsLM-IS257-like-mupA cassette. The IS257-like sequence of all MuH staphylococci showed two base pair substitutions and one base deletion compared with the sequence of IS257. The heterogeneous location of the mupA gene was caused by sequence deletion adjacent to the IS257-like sequence of the trsLM-IS257-like-mupA cassette construction, and the IS257-like sequence was found in all MuH staphylococci.

Investigation of beta-lactamase-producing multidrug-resistant Pseudomonas aeruginosa isolated from non-tertiary care hospitals in Korea.

A total of 2,280 nonduplicate clinical isolates of Pseudomonas aeruginosa, obtained nationwide from Korean non-tertiary care hospitals from 2002 to 2005, were identified and their susceptibilities to aminoglycosides, antipseudomonal penicillins, carbapenems, cephalosporins, monobactams, and quinolones were studied, together with their production of beta- lactamases. Using disk diffusion and minimum inhibitory concentration tests, it was found that 2.9% of isolates were multidrug-resistant (MDR) P. aeruginosa. An EDTA-disk synergy test, PCR amplification with specifically designed primers, and direct sequencing of the PCR products showed that the blaOXA-10, blaVIM-2, blaOXA-2, blaOXA-17, blaPER-1, blaSHV-12, and blaIMP-1 genes were carried by 34.3%, 26.9%, 3.0%, 3.0%, 1.5%, 1.5%, and 1.5% of 67 MDR P. aeruginosa isolates, respectively. The prevalence of MDR P. aeruginosa was three-fold higher, compared with that from the United States. More than two types of beta-lactamase genes were carried by 10.4% of isolates. The most prevalent beta-lactamase genes were blaVIM-2 and blaOXA-10. This study is the first description of MDR P. aeruginosa from non-tertiary care hospitals in Korea and the coexistence of the blaOXA-10 gene with blaVIM-2, blaIMP-1, or blaPER-1 in these clinical isolates.

Clonal dissemination and mupA gene polymorphism of mupirocin-resistant Staphylococcus aureus isolates from long-term-care facilities in South Korea.

We identified 25 high-level mupirocin-resistant (MuH) and 21 low-level mupirocin-resistant (MuL) Staphylococcus aureus isolates from eight long-term-care facilities (LTCFs). The pulsed-field gel electrophoresis patterns of 19 MuH and 19 MuL isolates from two facilities were identical for 18 and 15 isolates, respectively. The most predominant mupA restriction fragment length polymorphism type was found in 21 MuH isolates. We conclude that clonal transmission of MuH and MuL S. aureus strains occurred in these LTCFs. This is the first report of clonal transfer of mupirocin resistance in LTCFs.