Department of Microbiology, Nicolaus Copernicus University, Collegium Medicum , Bydgoszcz, Poland.
Arch Med Sci. 2012 Dec 20;8(6):993-7. doi: 10.5114/aoms.2012.32404. Epub 2012 Dec 19.
Twenty-eight isolates of extended-spectrum β-lactamase (ESBL)-positive Klebsiella pneumoniae were studied.
The strains were cultured from different clinical specimens obtained from children hospitalised at the University Hospital in Bydgoszcz. Seventeen strains were isolated from colonization and eleven from clinical infection. Isolation and identification of bacteria were performed using routine methods at the clinical microbiology laboratory. Production of ESBL was assessed using the double disk synergy test. The susceptibility to imipenem and tigecycline was tested by the Etest. The susceptibility to gentamicin and ciprofloxacin was tested by the agar dilution method. The genomic DNA was extracted from the strains separated by pulsed-field gel electrophoresis (PFGE) after digesting with XbaI endonuclease.
Among analysed K. pneumoniae strains all were susceptible to imipenem, 21 (75.0%) were susceptible to tigecycline, 14 (50.0%) to gentamicin and 5 (17.9%) to ciprofloxacin. Molecular typing results revealed a great genetic diversity among K. pneumoniae isolates. All repeated PFGE patterns were detected in seven K. pneumoniae isolates. Among identical K. pneumoniae strains four susceptibility patterns were detected.
The results of the study suggest that establishing strains' similarity in epidemiological investigations should be based on results obtained by several methods, and that each phenotyping method should be complemented with genetic research.
研究了 28 株产超广谱β-内酰胺酶(ESBL)的肺炎克雷伯菌。
这些菌株是从比得哥什大学医院住院儿童的不同临床标本中培养出来的。17 株从定植中分离,11 株从临床感染中分离。细菌的分离和鉴定在临床微生物学实验室使用常规方法进行。使用双碟协同试验评估 ESBL 的产生。使用 Etest 测试亚胺培南和替加环素的敏感性。使用琼脂稀释法测试庆大霉素和环丙沙星的敏感性。通过 XbaI 内切酶消化,从脉冲场凝胶电泳(PFGE)分离的菌株中提取基因组 DNA。
在所分析的肺炎克雷伯菌菌株中,所有菌株均对亚胺培南敏感,21 株(75.0%)对替加环素敏感,14 株(50.0%)对庆大霉素敏感,5 株(17.9%)对环丙沙星敏感。分子分型结果显示肺炎克雷伯菌分离株具有很大的遗传多样性。在 7 株肺炎克雷伯菌中均检测到相同的 PFGE 模式。在相同的肺炎克雷伯菌菌株中,检测到 4 种药敏模式。
研究结果表明,在流行病学调查中建立菌株相似性应基于几种方法的结果,并且每种表型方法都应辅以遗传研究。