Antibiotyping, RAPD- and ERIC-PCR fingerprinting of Klebsiella pneumoniae clinical isolates at a tertiary reference hospital in Denpasar, Bali, Indonesia.

IF 1.3 Q4 MICROBIOLOGY Iranian Journal of Microbiology Pub Date : 2024-06-01 DOI:10.18502/ijm.v16i3.15761
Ni Nengah Dwi Fatmawati, Felicia Aviana, Ronny Maharianto, Gede Ngurah Rsi Suwardana, Ni Made Adi Tarini, I Nengah Sujaya
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Abstract

Background and objectives: Klebsiella pneumoniae is a healthcare-associated infections agent and could be an extended spectrum β-lactamase (ESBL) producer. Understanding the transmission of this bacterium in a hospital setting needs accurate typing methods. An antibiogram is used to detect the resistance pattern of the isolates. Random Amplified Polymorphic DNA (RAPD) and Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR are rapid, technically simple, and easy-to-interpret DNA typing methods. This study aimed to evaluate the use of antibiotyping, RAPD-, and ERIC-PCR to investigate the heterogeneity of K. pneumoniae isolated from clinical specimens.

Materials and methods: The antibiograms of 46 K. pneumoniae clinical isolates were determined by Vitek® 2 Compact. All isolates underwent RAPD-PCR using AP4 primer and ERIC-PCR using ERIC-2 primer. The dendrogram was generated using the GelJ software and analyzed to determine its similarity. The analysis of antibiogram and the molecular typing diversity index was calculated using the formula of the Simpson's diversity index.

Results: About 71.7% of the isolates were ESBL-producers, and more than 80% of isolates were susceptible to amikacin, ertapenem, and meropenem. The antibiotyping produced 32 diverse types with DI = 0.964. In addition, the RAPD-PCR produced 47 different types with DI = 1, while ERIC-PCR was 46 (DI=0.999).

Conclusion: Antibiotyping, RAPD- and ERIC-PCR showed powerful discrimination power among the isolates, supported the diversity of K. pneumoniae isolates in current study. These combination could be promising tools for clonal relationship determination, including in tracking the transmission of the outbreak's agent in hospital setting.

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印度尼西亚巴厘岛登巴萨一家三级参考医院对肺炎克雷伯氏菌临床分离物进行抗生素鉴定、RAPD 和 ERIC-PCR 指纹图谱分析。
背景和目的:肺炎克雷伯氏菌是一种医疗相关性感染病原体,可能会产生广谱β-内酰胺酶(ESBL)。了解这种细菌在医院环境中的传播情况需要准确的分型方法。抗生素图谱可用于检测分离菌的耐药性模式。随机扩增多态 DNA (RAPD) 和肠杆菌重复基因间共识 (ERIC)-PCR 是快速、技术简单且易于解释的 DNA 分型方法。本研究旨在评估抗生素鉴定、RAPD 和 ERIC-PCR 在研究从临床标本中分离的肺炎克雷伯菌异质性方面的应用:采用 Vitek® 2 Compact 测定了 46 株肺炎克雷伯菌临床分离株的抗生素图谱。使用 AP4 引物对所有分离株进行 RAPD-PCR 检测,使用 ERIC-2 引物对所有分离株进行 ERIC-PCR 检测。使用 GelJ 软件生成树枝状图并进行分析,以确定其相似性。使用辛普森多样性指数公式计算抗生素图谱和分子分型多样性指数:结果:约71.7%的分离株产ESBL,80%以上的分离株对阿米卡星、厄他培南和美罗培南敏感。抗生素鉴定产生了 32 种不同类型,DI = 0.964。此外,RAPD-PCR 产生了 47 种不同类型,DI = 1,而 ERIC-PCR 为 46 种(DI=0.999):结论:抗生素鉴定、RAPD-PCR 和 ERIC-PCR 在分离株之间显示出强大的区分能力,支持了本次研究中肺炎克雷伯菌分离株的多样性。这些组合可能成为确定克隆关系的有效工具,包括追踪疫情病原体在医院环境中的传播。
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来源期刊
CiteScore
2.40
自引率
7.10%
发文量
96
审稿时长
12 weeks
期刊介绍: The Iranian Journal of Microbiology (IJM) is an international, multi-disciplinary, peer-reviewed journal that provides rapid publication of the most advanced scientific research in the areas of basic and applied research on bacteria and other micro-organisms, including bacteria, viruses, yeasts, fungi, microalgae, and protozoa concerning the development of tools for diagnosis and disease control, epidemiology, antimicrobial agents, clinical microbiology, immunology, Genetics, Genomics and Molecular Biology. Contributions may be in the form of original research papers, review articles, short communications, case reports, technical reports, and letters to the Editor. Research findings must be novel and the original data must be available for review by the Editors, if necessary. Studies that are preliminary, of weak originality or merely descriptive as well as negative results are not appropriate for the journal. Papers considered for publication must be unpublished work (except in an abstract form) that is not under consideration for publication anywhere else, and all co-authors should have agreed to the submission. Manuscripts should be written in English.
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