Área Bioquímica y Biología Molecular, Universidad de La Rioja, Madre de Dios 51, 26006 Logroño, Spain.
Int J Antimicrob Agents. 2012 Oct;40(4):306-12. doi: 10.1016/j.ijantimicag.2012.06.009. Epub 2012 Aug 14.
Nine staphylococcal strains of human and animal origin with a lincomycin-resistant/erythromycin-susceptible phenotype and carrying vga genes were characterised to determine the genetic elements involved in the dissemination of these uncommon resistance genes. These strains were typed by multilocus sequence typing (MLST), staphylococcal cassette chromosome mec (SCCmec) and/or spa typing. Antimicrobial susceptibility was studied by disk diffusion and agar dilution methods. Presence of the genes lnu(A), lnu(B), vga(A), vga(A)v, vga(B), vga(C), vga(E), lsa(B) and cfr was studied by PCR. Transformation experiments were carried out in all strains, and the plasmid or chromosomal gene location was determined by Southern blot analysis. Genetic environments of the vga genes were analysed by PCR mapping or inverse PCR and sequencing. Five meticillin-resistant Staphylococcus aureus (MRSA) ST398 strains and three Staphylococcus epidermidis strains harboured the gene vga(A), and one MRSA-ST8 strain contained the gene vga(A)v. One MRSA-ST398 strain, which also contained the gene lnu(A), showed the highest minimum inhibitory concentration (MIC) to lincomycin. The vga(A)v-positive strain presented lower MIC values than the vga(A)-positive strains. Presence of the pVGA plasmid was confirmed in two MRSA-ST398 strains. Four novel vga(A)-carrying plasmids were detected: pUR2355 (in two MRSA and one meticillin-susceptible S. epidermidis); pUR4128 (one MRSA); pUR3036 [one meticillin-resistant S. epidermidis (MRSE)]; and pUR3937 (one MRSE). The plasmid pUR4128 was very similar to pUR2355. Plasmids pUR3036 and pUR3937 were related and were very similar to plasmid pSE-12228-06. The gene vga(A)v was located in a transposon analogous to Tn5406. Therefore, four novel vga(A)-carrying plasmids and a variant of Tn5406 were identified in this study.
九株具有林可霉素耐药/红霉素敏感表型的人源和动物源葡萄球菌分离株,均携带 vga 基因,这些分离株的特征在于确定这些不常见耐药基因传播所涉及的遗传元件。这些菌株通过多位点序列分型(MLST)、葡萄球菌盒式染色体 mec(SCCmec)和/或 spa 分型进行分型。通过纸片扩散法和琼脂稀释法研究了抗生素敏感性。通过 PCR 研究了 lnu(A)、lnu(B)、vga(A)、vga(A)v、vga(B)、vga(C)、vga(E)、lsa(B)和 cfr 基因的存在情况。在所有菌株中进行了转化实验,并通过 Southern blot 分析确定了质粒或染色体基因位置。通过 PCR 图谱分析或反向 PCR 和测序分析了 vga 基因的遗传环境。五株耐甲氧西林金黄色葡萄球菌(MRSA)ST398 株和三株表皮葡萄球菌株携带 vga(A)基因,一株 MRSA-ST8 株携带 vga(A)v 基因。一株同时携带 lnu(A)基因的 MRSA-ST398 株对林可霉素的最小抑菌浓度(MIC)最高。vga(A)v 阳性株的 MIC 值低于 vga(A)阳性株。在两株 MRSA-ST398 株中证实了 pVGA 质粒的存在。检测到四种新型 vga(A)携带质粒:pUR2355(两株 MRSA 和一株耐甲氧西林表皮葡萄球菌);pUR4128(一株 MRSA);pUR3036[一株耐甲氧西林表皮葡萄球菌(MRSE)];和 pUR3937(一株 MRSE)。pUR4128 质粒与 pUR2355 非常相似。pUR3036 和 pUR3937 质粒相关,与质粒 pSE-12228-06 非常相似。基因 vga(A)v 位于类似于 Tn5406 的转座子中。因此,本研究鉴定了四种新型 vga(A)携带质粒和 Tn5406 的变体。