Laboratorio de Ciencias Omicas, Facultad de Ciencias de La Salud, Universidad Espíritu Santo, Samborondón, Ecuador.
Hospital de Niños Dr. Roberto Gilbert E. Junta de Beneficencia de Guayaquil, Guayaquil, Ecuador.
BMC Genomics. 2024 Oct 4;25(1):928. doi: 10.1186/s12864-024-10835-9.
Klebsiella pneumoniae is the major cause of nosocomial infections worldwide and is related to a worsening increase in Multidrug-Resistant Bacteria (MDR) and virulence genes that seriously affect immunosuppressed patients, long-stay intensive care patients, elderly individuals, and children. Whole-Genome Sequencing (WGS) has resulted in a useful strategy for characterizing the genomic components of clinically important bacteria, such as K. pneumoniae, enabling them to monitor genetic changes and understand transmission, highlighting the risk of dissemination of resistance and virulence associated genes in hospitals. In this study, we report on WGS 14 clinical isolates of K. pneumoniae from a pediatric hospital biobank of Guayaquil, Ecuador.
The main findings revealed pronounced genetic heterogeneity among the isolates. Multilocus sequencing type ST45 was the predominant lineage among non-KPC isolates, whereas ST629 was found more frequently among KPC isolates. Phylogenetic analysis suggested local transmission dynamics. Comparative genomic analysis revealed a core set of 3511 conserved genes and an open pangenome in neonatal isolates. The diversity of MLSTs and capsular types, and the high genetic diversity among these isolates indicate high intraspecific variability. In terms of virulence factors, we identified genes associated with adherence, biofilm formation, immune evasion, secretion systems, multidrug efflux pump transporters, and a notably high number of genes related to iron uptake. A large number of these genes were detected in the ST45 isolate, whereas iron uptake yersiniabactin genes were found exclusively in the non-KPC isolates. We observed high resistance to commonly used antibiotics and determined that these isolates exhibited multidrug resistance including β-lactams, aminoglycosides, fluoroquinolones, quinolones, trimetropins, fosfomycin and macrolides; additionally, resistance-associated point mutations and cross-resistance genes were identified in all the isolates. We also report the first K. pneumoniae KPC-3 gene producers in Ecuador.
Our WGS results for clinical isolates highlight the importance of MDR in neonatal K. pneumoniae infections and their genetic diversity. WGS will be an imperative strategy for the surveillance of K. pneumoniae in Ecuador, and will contribute to identifying effective treatment strategies for K. pneumoniae infections in critical units in patients at stratified risk.
肺炎克雷伯菌是全球导致医院感染的主要原因,与多药耐药菌(MDR)和毒力基因的日益增加有关,这些基因严重影响免疫抑制患者、长期入住重症监护病房的患者、老年人和儿童。全基因组测序(WGS)已成为一种有用的策略,可用于描述肺炎克雷伯菌等临床重要细菌的基因组成分,使我们能够监测遗传变化并了解传播情况,突出了医院中耐药性和毒力相关基因传播的风险。在这项研究中,我们报告了来自厄瓜多尔瓜亚基尔一家儿科医院生物库的 14 株肺炎克雷伯菌的 WGS 结果。
主要发现揭示了这些分离株之间明显的遗传异质性。非 KPC 分离株中主要的多位点序列型为 ST45,而 KPC 分离株中更常见的是 ST629。系统发育分析表明存在局部传播动态。比较基因组分析显示新生儿分离株中有一个核心的 3511 个保守基因和一个开放的泛基因组。这些分离株的 MLST 型和荚膜型多样性以及遗传多样性高表明种内变异大。在毒力因子方面,我们鉴定了与粘附、生物膜形成、免疫逃避、分泌系统、多药外排泵转运体相关的基因,以及与铁摄取相关的大量基因。这些基因中的许多在 ST45 分离株中被检测到,而铁摄取耶尔森氏菌素基因仅在非 KPC 分离株中被发现。我们观察到对常用抗生素的高度耐药性,并确定这些分离株表现出包括β-内酰胺类、氨基糖苷类、氟喹诺酮类、喹诺酮类、三甲基嘧啶、磷霉素和大环内酯类在内的多种药物耐药性;此外,所有分离株均发现了耐药相关点突变和交叉耐药基因。我们还报告了厄瓜多尔首例肺炎克雷伯菌 KPC-3 基因产生菌。
我们对临床分离株的 WGS 结果突出了新生儿肺炎克雷伯菌感染中 MDR 的重要性及其遗传多样性。WGS 将成为厄瓜多尔监测肺炎克雷伯菌的必要策略,并有助于确定对高危患者重症监护病房中肺炎克雷伯菌感染的有效治疗策略。