Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales

İpek Koçer; Tekin Karsligil; Mustafa Sağlam; Uğur Arslanyürekli; İhsan Deveci; Emel Şahin

doi:10.3855/jidc.17096

Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales

Authors

İpek Koçer Department of Medical Microbiology, Sanko University School of Medicine, Gaziantep, Turkey https://orcid.org/0000-0002-0631-6415
Tekin Karsligil Department of Medical Microbiology, Gaziantep University School of Medicine, Gaziantep, Turkey https://orcid.org/0000-0001-7672-3625
Mustafa Sağlam Department of Medical Microbiology, Gaziantep University School of Medicine, Gaziantep, Turkey https://orcid.org/0000-0002-0479-3250
Uğur Arslanyürekli Department of Medical Biology, Gaziantep University School of Medicine, Gaziantep, Turkey
İhsan Deveci Department of Medical Biology, Gaziantep University School of Medicine, Gaziantep, Turkey
Emel Şahin Department of Medical Biology, Gaziantep University School of Medicine, Gaziantep, Turkey https://orcid.org/0000-0002-3233-8044

DOI:

https://doi.org/10.3855/jidc.17096

Keywords:

Enterobacteriales, carbapenemase, flow cytometry, meropenem

Abstract

Introduction: Infections due to carbapenem-resistant Enterobacteriales, which have increased worldwide in recent years, cause concern. This study aimed to rapidly detect carbapenemase gene region by using flow cytometry in Enterobacteriales isolates and to evaluate its efficiency and susceptibility by comparing it with polymerase chain reaction (PCR).

Methodology: In the study, 21 isolates obtained from the blood cultures of patients hospitalized in intensive care units and found to intermediate or resistant to at least one carbapenem in the automated system, and 14 isolates belonging to the carbapenem-susceptible Enterobacteriales family were included. Carbapenemase gene regions were investigated by PCR after their susceptibility was determined by disk diffusion method. Bacterial suspensions were treated with meropenem + specific carbapenemase inhibitors (EDTA or APBA) and Temocillin and stained with thiazole orange (TO) and propidium iodide (PI) to show dead/live cell differentiation. Dead/live cell percentages were calculated after reading on the flow cytometer device.

Results: In the ROC analysis of the flow cytometry method, the cut-off value, specificity, and susceptibility of PI staining rates for meropenem were found as 14.37%, 100%, and 65%, respectively. It was found that the flow cytometry method was well-compatible with PCR in the detection of the carbapenemase gene region.

Conclusions: Flow cytometry will continue to be a promising method for the detection of antimicrobial susceptibility and resistance due to its rapid analysis of many cells and its high compatibility with PCR results.

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Published

2023-05-31

How to Cite

Koçer İpek, Karsligil T, Sağlam M, Arslanyürekli U, Deveci İhsan, Şahin E (2023) Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales. J Infect Dev Ctries 17:635–642. doi: 10.3855/jidc.17096

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Issue

Vol. 17 No. 05: May 2023

Section

Original Articles

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