The objective of this study was to determine clinical-epidemiological characteristics of the patients and the genetic characteristics of carbapenemase KPC-3-producing Klebsiella pneumoniae isolates belonging to sequence type ST258. The eligible study population was all patients with isolates detected between October 2015 and March 2017. Clinical-epidemiological and microbiological data were gathered on risk factors associated with infection by this clone. Antimicrobial susceptibility was determined using MicroScan system and diffusion in agar. Genes encoding carbapenemases were detected using PCR and Sanger sequencing. The sequence type was assigned by MLST, and the genetic relationship among clinical isolates was determined by pulsed field electrophoresis and by analysis of the genetic environment. The study included 23 individuals with isolates of KPC-3/ST258; the mean age was 77 year, and mean stay pre-isolation was 32 days; 81% received empirical antimicrobial treatment. Isolates were only susceptible to gentamicin (CIM ≤ 2 mg/L), tigecycline (CIM ≤ 1 mg/L), and colistin (CIM ≤ 2 mg/L). The isolates belonged to ST258, with five pulse types or subgroups. All isolates showed amplification of KPC, which was identified as KPC-3 variant. Gene blaKPC-3 was flanked by insertion sequences Kpn6 and Kpn7 within Tn4401 transposon isoform a. We report, for the first time in Spain, an 18-month outbreak by KPC-3-producing ST258 K. pneumoniae. Its acquisition was associated with a history of antimicrobial therapy, with three treatment options, and with high mortality. The detection of different pulse types is attributable to different introductions of the clone in our setting, supporting the need for multi-resistant isolate surveillance studies.

中文翻译：

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通过危险因素和分子特征追踪格拉纳达（西班牙安达卢西亚）三级医院中产生KPC-3的ST-258肺炎克雷伯菌的暴发。

这项研究的目的是确定患者的临床流行病学特征以及属于序列类型ST258的产碳青霉烯酶KPC-3的肺炎克雷伯菌的遗传特征。符合条件的研究人群是2015年10月至2017年3月之间检测到的所有分离株患者。收集了该克隆与感染相关的危险因素的临床流行病学和微生物学数据。使用MicroScan系统和在琼脂中的扩散测定了抗生素敏感性。使用PCR和Sanger测序检测编码碳青霉烯酶的基因。通过MLST分配序列类型，并通过脉冲场电泳和遗传环境分析确定临床分离株之间的遗传关系。该研究包括23名分离出KPC-3 / ST258的个体。平均年龄为77岁，平均隔离前停留时间为32天；81％接受了经验性抗菌治疗。分离株仅对庆大霉素（CIM≤2 mg / L），替加环素（CIM≤1 mg / L）和粘菌素（CIM≤2 mg / L）敏感。分离物属于ST258，具有五个脉冲类型或子组。所有分离株均显示出KPC的扩增，已鉴定为KPC-3变体。基因blaKPC-3的侧翼是Tn4401转座子同种型a内的插入序列Kpn6和Kpn7。我们在西班牙首次报告了生产KPC-3的ST258肺炎克雷伯菌爆发18个月。它的获得与抗微生物治疗的历史，三种治疗选择以及高死亡率有关。