Department of Microbiology, Faculty of Medicine, and University Hospital in Pilsen, Charles University, Pilsen, Czech Republic.
Biomedical Center, Faculty of Medicine, Charles University, Pilsen, Czech Republic.
mSphere. 2020 Dec 9;5(6):e01008-20. doi: 10.1128/mSphere.01008-20.
The aim of this study was to report the characterization of the first -positive isolated from Czech hospitals. In 2019, one and four isolates were recovered from Czech hospitals. The production of carbapenemases was examined by a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) imipenem hydrolysis assay. Additionally, bacteria were screened for the presence of carbapenemase-encoding genes and plasmid-mediated colistin resistance genes by PCR. To define the genetic units carrying genes, the genomic DNAs of -carrying clinical isolates were sequenced on the PacBio Sequel I platform. Results showed that all isolates carried - and -like genes. Analysis of whole-genome sequencing (WGS) data revealed that all isolates carried -like alleles. Furthermore, the three sequence type 106 (ST106) isolates harbored the gene, while the ST764 and ST95 included Analysis of plasmid sequences showed that, in all isolates, was carried on IncHI2 plasmids. Additionally, at least one multidrug resistance (MDR) region was identified in each -carrying IncHI2 plasmid. The gene was found in the MDR regions of p48880_MCR_VIM and p51929_MCR_VIM. In the three remaining isolates, was localized on plasmids (∼55 kb) exhibiting -like sequences 99% identical to the respective gene of pKPC-CAV1193. In conclusion, to the best of our knowledge, these 5 isolates were the first -positive bacteria of clinical origin identified in the Czech Republic. Additionally, the carriage of the on pKPC-CAV1193-like plasmids is described for the first time. Thus, our findings underline the ongoing evolution of mobile elements implicated in the dissemination of clinically important resistance determinants. Infections caused by carbapenemase-producing bacteria have led to the revival of polymyxins as the "last-resort" antibiotic. Since 2016, several reports describing the presence of plasmid-mediated colistin resistance genes, , in different host species and geographic areas were published. Here, we report the first detection of carrying -9-like alleles isolated from Czech hospitals in 2019. Furthermore, the three ST106 isolates harbored , while the ST764 and ST95 isolates included Analysis of WGS data showed that, in all isolates, was carried on IncHI2 plasmids. was found in the MDR regions of IncHI2 plasmids, while was localized on pKPC-CAV1193-like plasmids, described here for the first time. These findings underline the ongoing evolution of mobile elements implicated in dissemination of clinically important resistance determinants. Thus, WGS characterization of MDR bacteria is crucial to unravel the mechanisms involved in dissemination of resistance mechanisms.
本研究旨在报告从捷克医院分离的第一株阳性菌的特征。2019 年，从捷克医院分离出一株肠杆菌科细菌和四株肺炎克雷伯菌。通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）法检测碳青霉烯酶的产生。此外，通过 PCR 筛选细菌中是否存在碳青霉烯酶编码基因和质粒介导的多粘菌素耐药基因。为了确定携带 基因的遗传单位，对携带 基因的临床分离株的基因组 DNA 进行了 PacBio Sequel I 平台测序。结果表明，所有分离株均携带 基因和 基因。全基因组测序（WGS）数据分析显示，所有分离株均携带 基因。此外，3 株 ST106 分离株携带 基因，而 ST764 和 ST95 株则携带 基因。质粒序列分析显示，所有分离株的 基因均位于 IncHI2 质粒上。此外，在每个携带 基因的 IncHI2 质粒中都鉴定到至少一个多药耐药（MDR）区域。基因位于 p48880_MCR_VIM 和 p51929_MCR_VIM 的 MDR 区域中。在其余 3 株分离株中， 基因定位于质粒上（约 55kb），与 pKPC-CAV1193 基因的序列相似性为 99%。总之，据我们所知，这些 5 株分离株是捷克共和国首次发现的临床来源的阳性菌。此外，首次描述了 基因位于 pKPC-CAV1193 样质粒上。因此，我们的研究结果强调了参与传播临床重要耐药决定因素的移动元件的持续进化。由产碳青霉烯酶细菌引起的感染导致多粘菌素作为“最后手段”抗生素的重新使用。自 2016 年以来，描述不同宿主物种和地理区域携带质粒介导的多粘菌素耐药基因 基因的报告陆续发表。在此，我们报告了 2019 年从捷克医院分离的携带 -9 样等位基因的 基因的首次检测。此外，3 株 ST106 分离株携带 基因，而 ST764 和 ST95 分离株则携带 基因。WGS 数据分析显示，所有分离株的 基因均位于 IncHI2 质粒上。基因位于 IncHI2 质粒的 MDR 区域，而 基因定位于 pKPC-CAV1193 样质粒上，这是首次报道。这些发现强调了参与传播临床重要耐药决定因素的移动元件的持续进化。因此，对多药耐药细菌进行 WGS 特征分析对于阐明耐药机制传播所涉及的机制至关重要。
