Abstract

BackgroundAcinetobacter baumannii is a cosmopolitan bacterium that is frequently reported from hospitalized patients, especially those patients who admitted in the intensive care unit. Recently, multiplex real-time PCR has been introduced for rapid detection of the resistance genes in clinical isolates of bacteria. The current study aimed to develop and evaluate multiplex real-time PCR to detect common resistance genes among clinical isolates of A. baumannii.ResultsMultiplex real-time PCR based on melting curve analysis showed different Tm corresponding to the amplified fragment consisted of 83.5 °C, 93.3 °C and 89.3 °C for blaIMP, blaVIM and blaNDM, respectively. Results of multiplex real-time PCR showed that the prevalence of blaIMP, blaVIM and blaNDM among the clinical isolates of A. baumannii were 5/128(3.9%), 9/128(7.03%) and 0/128(0%), respectively. Multiplex real-time PCR was able to simultaneously identify the resistance genes, while showed 100% concordance with the results of conventional PCR.ConclusionsThe current study showed that blaVIM, was the most prevalent MBL gene among the clinical isolates of A. baumannii while no amplification of blaNDM was seen. Multiplex real-time PCR can be sensitive and reliable technique for rapid detection of resistance genes in clinical isolates.

Highlights

  • Acinetobacter baumannii is a cosmopolitan bacterium that is frequently reported from hospitalized patients, especially those patients who admitted in the intensive care unit

  • Acinetobacter baumannii is known as one of the most common bacteria that is frequently found in the hospitalized patients in Intensive Care Unit (ICU) [1, 2]

  • In order to perform multiplex real-time PCR, A. baumannii isolates were selected from those strains that were previously phenotypically examined for piperacillin-tazobactam, ceftriaxon, ceftazidime, cefepime, imipenem, doripenemertapenem, cefotaxime, and ampicillin-sulbactam, using disc diffusion (Mast Co., UK) regarding to CLSI 2013 guidelines

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Summary

Introduction

Acinetobacter baumannii is a cosmopolitan bacterium that is frequently reported from hospitalized patients, especially those patients who admitted in the intensive care unit. The current study aimed to develop and evaluate multiplex real-time PCR to detect common resistance genes among clinical isolates of A. baumannii. Acinetobacter baumannii is known as one of the most common bacteria that is frequently found in the hospitalized patients in Intensive Care Unit (ICU) [1, 2]. Several reports of MBL-producing strains has led to increased numbers of studies on the prevalence as well as designing reliable methods for detection of the most prevalent MBL genes among clinical strains of A. baumannii [7, 15, 16]. Laboratory identification of the drug resistance genes is a pivotal step in the early assessment and management of hospital infections due to A. baumannii, in ICU patients

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