Laboratoire de Biochimie et de Microbiologie Appliquée, Département de Biochimie, Université Badji Mokhtar-Annaba, Annaba, Algérie.
Aix Marseille Univ, IRD, APHM, MEPHI, Marseille, France.
Microb Drug Resist. 2021 May;27(5):652-659. doi: 10.1089/mdr.2020.0080. Epub 2020 Sep 1.
The aim of this study was to characterize the molecular mechanisms of carbapenem resistance in isolated from the urology department of Annaba hospital, Algeria. Between January 2015 and September 2017, 14 carbapenem-resistant strains were isolated during routine surveillance work at Ibn Roched hospital of Annaba, Algeria, from the urology department. Theses strains were recovered, and carbapenem resistance mechanisms were investigated. The strains were identified by using matrix-assisted laser desorption and ionization time-of-flight mass spectrometry. Antibiotic susceptibility was assessed by using the Kirby-Bauer method, whereas minimum inhibitory concentration of imipenem/ertapenem and colistin was determined by Etest and broth microdilution methods, respectively. Carbapenem resistance determinants were studied by using PCR and sequencing methods and analyzed by BLAST against the Antibiotic Resistance Gene-ANNOTation (ARG-ANNOT) database. Clonal relationship of strains was performed by using multilocus sequence typing (MLST). Transferability of carbapenem resistance genes was assessed by conjugation and transformation experiments. Fourteen carbapenem-resistant isolates were found to be resistant to the eight β-lactam antibiotics tested (except to imipenem for two isolates). Carbapenemase production was positive for all isolates. Molecular characterization revealed that and genes were detected in 3 (21.4%) and 11 isolates (78.6%), respectively. Other β-lactamases genes were identified, including , , and . MLST revealed that the 14 isolates belonged to 2 different sequence types (STs), including ST101 (11 OXA-48-producing ) and ST258 (3 KPC-2-producing ). PCR amplifications for and carbapenemases genes performed on extracted plasmids, showed positive results, suggesting that both carbapenemase genes were probably borne by plasmids. We report here the first identification of KPC-2-producing ST258 in Algerian hospitals and an outbreak of OXA-48-producing isolates ST101 in the urology department of Ibn Roched hospital located in Annaba, Algeria.
本研究旨在探讨从阿尔及利亚安纳巴医院泌尿科分离的 中碳青霉烯类耐药的分子机制。2015 年 1 月至 2017 年 9 月，在阿尔及利亚安纳巴 Ibn Roched 医院的常规监测工作中，从泌尿科分离出 14 株碳青霉烯类耐药 株。回收这些菌株，研究碳青霉烯类耐药机制。采用基质辅助激光解吸电离飞行时间质谱法鉴定菌株。采用 Kirby-Bauer 法评估抗生素敏感性，采用 Etest 和肉汤微量稀释法分别测定亚胺培南/厄他培南和黏菌素的最小抑菌浓度。采用 PCR 和测序法研究碳青霉烯类耐药决定因子，并与抗生素耐药基因注释（ARG-ANNOT）数据库进行 BLAST 分析。采用多位点序列分型（MLST）分析菌株的克隆关系。通过接合和转化实验评估碳青霉烯类耐药基因的可转移性。发现 14 株碳青霉烯类耐药 株对 8 种β-内酰胺类抗生素（两种菌株对亚胺培南除外）均耐药。所有分离株均产碳青霉烯酶。分子特征分析显示，3 株（21.4%）和 11 株（78.6%）分离株分别检出 基因和 基因。还鉴定出其他β-内酰胺酶基因，包括 、 、和 。MLST 显示，14 株分离株属于 2 种不同的序列类型（STs），包括 11 株产 OXA-48 的 ST101 和 3 株产 KPC-2 的 ST258。对提取质粒进行 基因和 基因的 PCR 扩增，结果阳性，提示两种碳青霉烯酶基因可能均由质粒携带。我们首次在阿尔及利亚医院发现 KPC-2 产 株 ST258，并在位于阿尔及利亚安纳巴的 Ibn Roched 医院泌尿科发现 OXA-48 产 株 ST101 的爆发。
