Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales
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...
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The Journal of Infection in Developing Countries
2023-05-01
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| Series: | Journal of Infection in Developing Countries |
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| Online Access: | https://jidc.org/index.php/journal/article/view/17096 |
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| author | İpek Koçer Tekin Karsligil Mustafa Sağlam Uğur Arslanyürekli İhsan Deveci Emel Şahin |
| author_facet | İpek Koçer Tekin Karsligil Mustafa Sağlam Uğur Arslanyürekli İhsan Deveci Emel Şahin |
| author_sort | İpek Koçer |
| collection | DOAJ |
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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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| format | Article |
| id | doaj-art-3d70e638caf84b2c97f8e62dcf90eef4 |
| institution | Kabale University |
| issn | 1972-2680 |
| language | English |
| publishDate | 2023-05-01 |
| publisher | The Journal of Infection in Developing Countries |
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| series | Journal of Infection in Developing Countries |
| spelling | doaj-art-3d70e638caf84b2c97f8e62dcf90eef42025-08-20T03:52:43ZengThe Journal of Infection in Developing CountriesJournal of Infection in Developing Countries1972-26802023-05-01170510.3855/jidc.17096Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacterialesİpek Koçer0Tekin Karsligil1Mustafa Sağlam2Uğur Arslanyürekli3İhsan Deveci4Emel Şahin5Department of Medical Microbiology, Sanko University School of Medicine, Gaziantep, TurkeyDepartment of Medical Microbiology, Gaziantep University School of Medicine, Gaziantep, TurkeyDepartment of Medical Microbiology, Gaziantep University School of Medicine, Gaziantep, TurkeyDepartment of Medical Biology, Gaziantep University School of Medicine, Gaziantep, TurkeyDepartment of Medical Biology, Gaziantep University School of Medicine, Gaziantep, TurkeyDepartment of Medical Biology, Gaziantep University School of Medicine, Gaziantep, Turkey 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. https://jidc.org/index.php/journal/article/view/17096Enterobacterialescarbapenemaseflow cytometrymeropenem |
| spellingShingle | İpek Koçer Tekin Karsligil Mustafa Sağlam Uğur Arslanyürekli İhsan Deveci Emel Şahin Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales Journal of Infection in Developing Countries Enterobacteriales carbapenemase flow cytometry meropenem |
| title | Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales |
| title_full | Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales |
| title_fullStr | Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales |
| title_full_unstemmed | Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales |
| title_short | Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales |
| title_sort | evaluation of real time pcr and flow cytometry efficiency in rapid detection of carbapenemase producing enterobacteriales |
| topic | Enterobacteriales carbapenemase flow cytometry meropenem |
| url | https://jidc.org/index.php/journal/article/view/17096 |
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