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      Genetic profiling of extended-spectrum β-Lactamase and carbapenemase-producing Escherichia coli O157:H7 from clinical samples among diarrheal patients in Shashemene, Ethiopia

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          Abstract

          Background

          Escherichia coli (E. coli) O157:H7, associated with diarrhea, poses a global health risk. In Ethiopia, where diarrhea is common, there is limited knowledge about these resistant strains and a lack of data on Extended-Spectrum β-Lactamase (ESBL) and carbapenemase production. Understanding the prevalence of antimicrobial resistance genes associated with ESBL and carbapenems is crucial for addressing diarrheal disease. This study aimed to investigate the genetic profile of ESBL and carbapenemase coding gene carriage in E. coli O157:H7 from clinical stool samples and evaluate antimicrobial susceptibility patterns.

          Methods

          A total of twenty-nine bacterial isolates obtained from diarrheal patients were subjected to conventional culture and phenotypic (Kirby Bauer disc diffusion method) testing for antimicrobial resistance. Additionally, screening for the production of ESBL (combined disk method) and carbapenemase (modified carbapenem inactivation method) was conducted. Isolates that tested positive for ESBL and carbapenemase production were further analyzed, targeting five genes ( bla NDM, bla KPC, bla CTX−M, bla TEM, and bla SHV) associated with ESBL and carbapenemase production. Data analysis was performed using SPSS version 27.0, employing logistic regression and descriptive statistics.

          Results

          We analyzed a total of 27 isolates that were ESBL-positive and 12 isolates that were found to produce carbapenemase phenotypically. These isolates were obtained from clinical stool samples and (9/27) 33.3% of the isolates were from under five years children, predominantly from urban areas, and those that have contact with domestic animals. Genes coding ESBL were found in (19/27) 70.4% of the isolates, the most predominant being bla CTX−M and bla TEM. Eight isolates carried bla KPC, but none had bla NDM, while five isolates carried both bla CTX−M and bla TEM genes. bla SHV-carrying isolates showed phenotypic resistance to ampicillin and cephalosporins, while bla KPC-carrying isolates exhibited resistance to ampicillin, carbapenems, and tetracycline.

          Conclusion

          This study identifies a significant prevalence of multidrug resistance in E. coli O157:H7, which can be attributed to the presence of resistance genes coding for ESBL and carbapenem production. Key factors contributing to this resistance, such as urban environments, children under the age of five, and domestic animal ownership, have been emphasized. Additionally, this research underscores the urgent need for enhanced surveillance and targeted interventions to address this pressing public health concern.

          Supplementary Information

          The online version contains supplementary material available at 10.1186/s12879-025-10513-5.

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          Most cited references21

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          Multiplex PCR for detection of acquired carbapenemase genes.

          A rapid and reliable PCR-based technique was developed for detection of genes encoding carbapenemases belonging to different classes. Primers were designed to amplify the following 11 genes: bla(IMP), bla(VIM), bla(NDM), bla(SPM), bla(AIM), bla(DIM), bla(GIM), bla(SIM)bla(KPC), bla(BIC), and bla(OXA-48). Three different multiplex reaction mixtures were defined and evaluated for the detection of all these 11 genes. Using optimized conditions, each reaction mixture allowed to identify the respective genes, with PCR giving distinct amplicon sizes corresponding to the different genes for each mixture. We reported here a rapid and reliable technique for screening all clinically relevant carbapenemase genes. Copyright © 2011 Elsevier Inc. All rights reserved.
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            Global epidemiology of CTX-M β-lactamases: temporal and geographical shifts in genotype.

            Globally, rates of ESBL-producing Enterobacteriaceae are rising. We undertook a literature review, and present the temporal trends in blaCTX-M epidemiology, showing that blaCTX-M-15 and blaCTX-M-14 have displaced other genotypes in many parts of the world. Explanations for these changes can be attributed to: (i) horizontal gene transfer (HGT) of plasmids; (ii) successful Escherichia coli clones; (iii) ESBLs in food animals; (iv) the natural environment; and (v) human migration and access to basic sanitation. We also provide explanations for the changing epidemiology of blaCTX-M-2 and blaCTX-M-27. Modifiable anthropogenic factors, such as poor access to basic sanitary facilities, encourage the spread of blaCTX-M and other antimicrobial resistance (AMR) genes, such as blaNDM, blaKPC and mcr-1. We provide further justification for novel preventative and interventional strategies to reduce transmission of these AMR genes.
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              Heat treatment of bacteria: a simple method of DNA extraction for molecular techniques

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                Author and article information

                Contributors
                shimelisteshome35@gmail.com
                Journal
                BMC Infect Dis
                BMC Infect Dis
                BMC Infectious Diseases
                BioMed Central (London )
                1471-2334
                20 January 2025
                20 January 2025
                2025
                : 25
                : 90
                Affiliations
                [1 ]Department of Medical Laboratory Sciences, College of Health Sciences, Arsi University, ( https://ror.org/04s6kmw55) Asella, Ethiopia
                [2 ]Bacterial and Viral Disease Research Directorate, Armauer Hansen Research Institute, ( https://ror.org/05mfff588) Addis Ababa, Ethiopia
                [3 ]Department of Microbiology, Immunology and Parasitology, School of Medicine, College of Health Sciences, Addis Ababa University, ( https://ror.org/038b8e254) Addis Ababa, Ethiopia
                [4 ]Clinical, Assessment, Regulatory, Evaluation (CARE) Unit, International Vaccine Institute, ( https://ror.org/02yfanq70) Seoul, Republic of Korea
                [5 ]Clinical Trials Directorate, Armauer Hansen Research Institute, ( https://ror.org/05mfff588) Addis Ababa, Ethiopia
                [6 ]Yonsei University Graduate School of Public Health, ( https://ror.org/01wjejq96) Seoul, Republic of Korea
                Article
                10513
                10.1186/s12879-025-10513-5
                11748885
                39833755
                3cd5dd46-8e7e-41c3-bb26-b1a97ed6a21b
                © The Author(s) 2025

                Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

                History
                : 22 May 2024
                : 16 January 2025
                Categories
                Research
                Custom metadata
                © BioMed Central Ltd., part of Springer Nature 2025

                Infectious disease & Microbiology
                antimicrobial resistant genes,carbapenemase,esbl,e. coli o157:h7,ethiopia

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