Shahkolahi Shaghayegh, Shakibnia Pegah, Shahbazi Shahla, Sabzi Samira, Badmasti Farzad, Asadi Karam Mohammad Reza, Habibi Mehri
1Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran.
2Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran.
Acta Microbiol Immunol Hung. 2022 Sep 19;69(4):303-313. doi: 10.1556/030.2022.01808. Print 2022 Dec 6.
In the present study a total of 200 Klebsiella pneumoniae isolates were collected from patients with urinary tract infections (UTIs) in Tehran, Iran. Antibiotic resistance was determined by disk diffusion and broth dilution methods. Detection of extended-spectrum β-lactamases (ESBLs) and AmpCs was performed using phenotypic tests. Polymerase chain reaction (PCR) was applied to detect the ESBL, AmpC, and integron genes. Analysis of AmpC and cassette arrays of integron genes was performed using DNA sequencing. Plasmids were analyzed by PCR-based replicon typing and conjugation. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were applied to explore the genomic relatedness among the isolates. The highest levels of resistance were observed against ampicillin (100%), followed by piperacillin (57.5%), ceftazidime (46%), trimethoprim/sulfamethoxazole (44%), ciprofloxacin (32.5%), and imipenem (19%). Approximately, 66.5% of isolates harbored at least one of the beta-lactamase genes (blaTEM, blaSHV, blaCTX-M, and blaOXA-1). In addition, 22.5% of isolates carried at least one of the AmpC genes including blaDHA and blaCIT. Integron class I was the most prevalent integron among resistant isolates. According to the results of replicon typing, IncFII, IncL/M, and IncA/C were the most frequent replicons, respectively. All selected isolates were able to transfer blaCTX-M, also two isolates transferred the blaDHA-1 gene to Escherichia coli K12 through conjugation. Finally, 21 isolates were categorized into 4 pulsotypes and 11 unique clusters in PFGE. MLST identified ST147 and ST11 sequence types but ST147 was the most prevalent in the current study.
在本研究中,共从伊朗德黑兰患有尿路感染(UTIs)的患者中收集了200株肺炎克雷伯菌分离株。通过纸片扩散法和肉汤稀释法测定抗生素耐药性。使用表型试验检测超广谱β-内酰胺酶(ESBLs)和AmpC酶。应用聚合酶链反应(PCR)检测ESBL、AmpC和整合子基因。使用DNA测序对AmpC和整合子基因的盒式阵列进行分析。通过基于PCR的复制子分型和接合试验分析质粒。应用脉冲场凝胶电泳(PFGE)和多位点序列分型(MLST)来探索分离株之间的基因组相关性。观察到对氨苄西林的耐药水平最高(100%),其次是哌拉西林(57.5%)、头孢他啶(46%)、甲氧苄啶/磺胺甲恶唑(44%)、环丙沙星(32.5%)和亚胺培南(19%)。大约66.5%的分离株携带至少一种β-内酰胺酶基因(blaTEM、blaSHV、blaCTX-M和blaOXA-1)。此外,22.5%的分离株携带至少一种AmpC基因包括blaDHA和blaCIT。I类整合子是耐药分离株中最常见的整合子。根据复制子分型结果,IncFII、IncL/M和IncA/C分别是最常见的复制子。所有选定的分离株都能够转移blaCTX-M,也有两株分离株通过接合将blaDHA-1基因转移到大肠杆菌K12中。最后,21株分离株在PFGE中被分为4个脉冲型和11个独特的簇。MLST鉴定出ST147和ST11序列型,但ST147在本研究中最为普遍。