Department of Biology, Rasht Branch, Islamic Azad University, Rasht, Iran.
Int Microbiol. 2019 Dec;22(4):411-417. doi: 10.1007/s10123-019-00063-7. Epub 2019 Feb 18.
Glycopeptides, particularly the cell wall-acting antibiotic vancomycin, are the safest cure for methicillin-resistant Staphylococcus aureus. The aim of this study was to evaluate nonsusceptibility of clinical isolates of S. aureus to vancomycin and investigate mutations in vraSR, a cell wall synthesis regulator gene, in vancomycin-resistant strains. Susceptibility of 110 clinical strains of S. aureus to methicillin and vancomycin were determined using disc diffusion method and determination of minimum inhibitory concentration, respectively. Presence of mecA and vanA genes was determined by PCR. Determination of spa types and mutations of the vraSR gene in vancomycin nonsusceptible isolates were assessed by PCR-sequencing analyses. In total, 47 isolates (42.73%) were recognized as MRSA, three (2.73%) strains were resistant to vancomycin, and eight (7.27%) strains were vancomycin intermediates. The MIC of vancomycin was 4-64 μg/ml in these isolates. All vancomycin nonsusceptible S. aureus strains were mecA positive and one isolate was positive for the vanA gene. Spa type t030 was found as the most common type. In vraSR sequence analysis, all 11 vancomycin nonsusceptible isolates had the D59E mutation in the vraR and E45G in vraS genes. R117H, R121S, and R121I are the other identified missense mutations in the vraR gene. The identification of a high percentage of MRSA and presence of VRSA and VISA isolates is a serious warning about the treatment of future MRSA infections and reveals the need for new and effective therapeutic agents.
糖肽类,特别是作用于细胞壁的抗生素万古霉素,是治疗耐甲氧西林金黄色葡萄球菌(MRSA)的最安全方法。本研究旨在评估临床金黄色葡萄球菌分离株对万古霉素的非敏感性,并研究万古霉素耐药株中细胞壁合成调节基因 vraSR 的突变情况。采用纸片扩散法和最小抑菌浓度测定法分别测定 110 株临床金黄色葡萄球菌对甲氧西林和万古霉素的敏感性。采用 PCR 法检测 mecA 和 vanA 基因的存在。通过 PCR-测序分析评估万古霉素非敏感性分离株的 spa 型和 vraSR 基因突变情况。共有 47 株(42.73%)被鉴定为 MRSA,3 株(2.73%)菌株对万古霉素耐药,8 株(7.27%)菌株为万古霉素中介。这些分离株的万古霉素 MIC 为 4-64μg/ml。所有万古霉素非敏感性金黄色葡萄球菌菌株均为 mecA 阳性,1 株分离株对 vanA 基因呈阳性。t030 型 spa 被发现为最常见的类型。在 vraSR 序列分析中,所有 11 株万古霉素非敏感性分离株的 vraR 基因均存在 D59E 突变,vraS 基因存在 E45G 突变。vraR 基因中还发现了其他 3 种错义突变,即 R117H、R121S 和 R121I。鉴定出高比例的 MRSA 以及 VRSA 和 VISA 分离株是对未来 MRSA 感染治疗的严重警告,并表明需要新的有效治疗药物。