Epidemiological and Genetic Overview of the Klebsiella pneumoniae Carbapenemases (KPCs) in K. pneumoniae Isolated from the Clinical Samples in Iran

Narjes Mohammadi Bandari, H. Keyvani, Mohsen Zargar, M. Talebi, M. Zolfaghari
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引用次数: 5

Abstract

Background: The prevalence of carbapenem-resistant Enterobacteriaceae, especially Klebsiella pneumoniae carbapenemase (KPC), has been recently reported worldwide. Therefore, there is an indispensable need for precise and rapid detection of these carbapenemases. Objectives: This study aimed to propose an accurate and rapid method for detecting K. pneumoniae carbapenemase genes from clinical samples, using reverse transcription-polymerase chain reaction (RT-PCR) and to evaluate the expression of these genes in the presence of β-lactam antibiotic by real-time PCR assay.Methods: One hundred and eighty-one K. pneumoniae strains were collected from patients presenting to Firoozgar Hospital of Tehran, Iran. The strains were tested using the disk diffusion method, the modified Hodge test (MHT), and E-test minimum inhibitory concentration (MIC). Next, reverse transcription-PCR method was applied for the identification of bla OXA-23 and bla OXA-48 genes. Finally, expression of genes was measured by real-time PCR assay in the presence and absence of β-lactam antibiotic.Results: Phenotypic testing also showed a high level of antibiotic resistance, while the genotypic methods indicated the presence and expression of carbapenemase genes.Conclusions: The findings suggest revisions in the current antibiotic therapy protocols, considering the high expression level of resistant carbapenemases to K. pneumoniae strains.
伊朗临床分离肺炎克雷伯菌碳青霉烯酶的流行病学和遗传学研究
背景:碳青霉烯耐药肠杆菌科,特别是肺炎克雷伯菌碳青霉烯酶(KPC)的流行,最近已在世界范围内报道。因此,对这些碳青霉烯酶进行精确、快速的检测是必不可少的。目的:本研究旨在建立一种准确、快速的临床样品中肺炎克雷伯菌碳青霉烯酶基因的逆转录聚合酶链反应(RT-PCR)检测方法,并通过实时荧光定量PCR检测这些基因在β-内酰胺类抗生素存在下的表达情况。方法:收集伊朗德黑兰Firoozgar医院就诊患者181株肺炎克雷伯菌。采用纸片扩散法、改良霍奇试验(MHT)和e -试验最低抑菌浓度(MIC)对菌株进行检测。接下来,应用逆转录- pcr方法对bla OXA-23和bla OXA-48基因进行鉴定。最后,采用实时荧光定量PCR法检测β-内酰胺抗生素存在和不存在情况下基因的表达。结果:表型检测也显示出高水平的抗生素耐药性,而基因分型方法表明碳青霉烯酶基因的存在和表达。结论:考虑到耐药碳青霉烯酶对肺炎克雷伯菌菌株的高表达水平,研究结果建议对目前的抗生素治疗方案进行修订。
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