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Characterization of Plasmid-Mediated AmpC and Carbapenemases among Iranain Nosocomial Isolates of Klebsiella pneumoniae Using Phenotyping and Genotyping Methods

机译:利用表型和基因分型方法表征肺炎克雷伯菌的伊朗分离株中质粒介导的AmpC和碳青霉烯酶。

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摘要

textabstractObjectives: Plasmid-mediated AmpC β-lactamases (PMABLs) and carbapenemases are emerging groups of antimicrobial-resistance determinants. The aims of the study were to evaluate the occurrence of PMABLs and carbapenemases in clinical isolates of Klebsiella pneumoniae and compare the test performance of various phenotypic methods for detection of these enzymes in Iran. Methods: A total of 100 K. pneumoniae isolates were collected from clinical specimens obtained in Valiasr Hospital. AmpC production in all isolates was determined using the AmpC disk test, the cephamycin Hodge test, the AmpC Etest, and the boronic acid combined-disk test. In addition, carbapenemase production was determined using the modified Hodge test, the EDTA disk synergy test, and the boronic acid combined-disk test. The performances of various phenotypic methods were evaluated by the comparison of their results with polymerase chain reaction (PCR) method as the gold standard. Results: Of the 100 isolates, 19 (19%) were demonstrated to harbor the PMABL-resistance gene by the multiplex PCR method. The PCR result indicated the presence of carbapenemase genes in 12 isolates. The performance of various phenotypic tests carried out for detection of carbapenemase-producing isolates varied widely, ranging in sensitivity from 30% to 100% and in specificity from 90.8% to 100%. Conclusion: This is the first report of MOX-type AmpC β-lactamase and blaGES in K. pneumoniae in Iran. A comparison of the phenotypic methods showed that a combination of cefoxitin plus boronic acid is optimal for detecting plasmid-mediated AmpC enzymes in K. pneumoniae, whereas the implementation of molecular methods is often complex, requires specially trained personnel, and is associated with higher costs.
机译:目的:质粒介导的AmpCβ-内酰胺酶(PMABLs)和碳青霉烯酶是新兴的抗药性决定因素。该研究的目的是评估在肺炎克雷伯菌的临床分离株中PMABLs和碳青霉烯酶的存在,并比较在伊朗检测这些酶的各种表型方法的测试性能。方法:从Valiasr医院获得的临床标本中共收集到100株肺炎克雷伯菌。使用AmpC圆盘测试,头孢菌素Hodge测试,AmpC Etest和硼酸联合圆盘测试确定所有分离株中AmpC的产量。此外,使用改良的Hodge试验,EDTA盘协同试验和硼酸组合盘试验确定了碳青霉烯酶的生产。以聚合酶链反应(PCR)法为金标准,通过比较它们的结果来评估各种表型方法的性能。结果:通过多重PCR方法,在100株分离株中,有19株(占19%)具有PMABL抗性基因。 PCR结果表明在12个分离物中存在碳青霉烯酶基因。用于检测产生碳青霉烯酶的分离物的各种表型测试的性能差异很大,灵敏度范围为30%至100%,特异性范围为90.8%至100%。结论:这是伊朗肺炎克雷伯菌中MOX型AmpCβ-内酰胺酶和blaGES的首次报道。表型方法的比较表明,头孢西丁和硼酸的组合是检测肺炎克雷伯菌中质粒介导的AmpC酶的最佳方法,而分子方法的实施通常很复杂,需要经过专门培训的人员,并且成本较高。

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