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Transcriptional expression of secondary resistance genes ccdB and repA2 is enhanced in presence of cephalosporin and carbapenem in Escherichia coli

BACKGROUND: The issue of carbapenem resistance in E.coli is very concerning and it is speculated that cumulative effect of both primary resistance genes and secondary resistance genes that act as helper to the primary resistance genes are the reason behind their aggravation. Therefore, here we attem...

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Autores principales: Baishya, Somorita, Deshamukhya, Chandrayee, Wangkheimayum, Jayalaxmi, Das, Bhaskar Jyoti, Anbarasu, Anand, Talukdar, Anupam Das, Bhattacharjee, Amitabha, Choudhury, Manabendra Dutta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7941987/
https://www.ncbi.nlm.nih.gov/pubmed/33750290
http://dx.doi.org/10.1186/s12866-021-02136-y
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author Baishya, Somorita
Deshamukhya, Chandrayee
Wangkheimayum, Jayalaxmi
Das, Bhaskar Jyoti
Anbarasu, Anand
Talukdar, Anupam Das
Bhattacharjee, Amitabha
Choudhury, Manabendra Dutta
author_facet Baishya, Somorita
Deshamukhya, Chandrayee
Wangkheimayum, Jayalaxmi
Das, Bhaskar Jyoti
Anbarasu, Anand
Talukdar, Anupam Das
Bhattacharjee, Amitabha
Choudhury, Manabendra Dutta
author_sort Baishya, Somorita
collection PubMed
description BACKGROUND: The issue of carbapenem resistance in E.coli is very concerning and it is speculated that cumulative effect of both primary resistance genes and secondary resistance genes that act as helper to the primary resistance genes are the reason behind their aggravation. Therefore, here we attempted to find the role of two secondary resistance genes (SRG) ccdB and repA2 in carbapenem resistance in E. coli (CRE). In this context influential genes belonging to secondary resistome that act as helper to the primary resistance genes like bla(NDM) and bla(CTX-M) in aggravating β-lactam resistance were selected from an earlier reported in silico study. Transcriptional expression of the selected genes in clinical isolates of E.coli that were discretely harboring bla(NDM-1), bla(NDM-4), bla(NDM-5), bla(NDM-7) and bla(CTX-M-15) with and without carbapenem and cephalosporin stress (2 μg/ml) was determined by real time PCR. Cured mutants sets that were lacking (i) primary resistance genes, (ii) secondary resistance genes and (iii) both primary and secondary resistance genes were prepared by SDS treatment. These sets were then subjected to antibiotic susceptibility testing by Kirby Bauer disc diffusion method. RESULTS: Out of the 21 genes reported in the in silico study, 2 genes viz. repA2 and ccdB were selected for transcriptional expression analysis. repA2, coding replication regulatory protein, was downregulated in response to carbapenems and cephalosporins. ccdB, coding for plasmid maintenance protein, was also downregulated in response to carbapenems except imipenem and cephalosporins. Following plasmid elimination assay increase in diameter of zone of inhibition under stress of both antibiotics was observed as compared to uncured control hinting at the reversion of antibiotic susceptibility by the-then resistant bacteria. CONCLUSION: SRGs repA2 and ccdB help sustenance of bla(NDM) and bla(CTX-M) under carbapenem and cephalosporin stress. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-021-02136-y.
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spelling pubmed-79419872021-03-10 Transcriptional expression of secondary resistance genes ccdB and repA2 is enhanced in presence of cephalosporin and carbapenem in Escherichia coli Baishya, Somorita Deshamukhya, Chandrayee Wangkheimayum, Jayalaxmi Das, Bhaskar Jyoti Anbarasu, Anand Talukdar, Anupam Das Bhattacharjee, Amitabha Choudhury, Manabendra Dutta BMC Microbiol Research Article BACKGROUND: The issue of carbapenem resistance in E.coli is very concerning and it is speculated that cumulative effect of both primary resistance genes and secondary resistance genes that act as helper to the primary resistance genes are the reason behind their aggravation. Therefore, here we attempted to find the role of two secondary resistance genes (SRG) ccdB and repA2 in carbapenem resistance in E. coli (CRE). In this context influential genes belonging to secondary resistome that act as helper to the primary resistance genes like bla(NDM) and bla(CTX-M) in aggravating β-lactam resistance were selected from an earlier reported in silico study. Transcriptional expression of the selected genes in clinical isolates of E.coli that were discretely harboring bla(NDM-1), bla(NDM-4), bla(NDM-5), bla(NDM-7) and bla(CTX-M-15) with and without carbapenem and cephalosporin stress (2 μg/ml) was determined by real time PCR. Cured mutants sets that were lacking (i) primary resistance genes, (ii) secondary resistance genes and (iii) both primary and secondary resistance genes were prepared by SDS treatment. These sets were then subjected to antibiotic susceptibility testing by Kirby Bauer disc diffusion method. RESULTS: Out of the 21 genes reported in the in silico study, 2 genes viz. repA2 and ccdB were selected for transcriptional expression analysis. repA2, coding replication regulatory protein, was downregulated in response to carbapenems and cephalosporins. ccdB, coding for plasmid maintenance protein, was also downregulated in response to carbapenems except imipenem and cephalosporins. Following plasmid elimination assay increase in diameter of zone of inhibition under stress of both antibiotics was observed as compared to uncured control hinting at the reversion of antibiotic susceptibility by the-then resistant bacteria. CONCLUSION: SRGs repA2 and ccdB help sustenance of bla(NDM) and bla(CTX-M) under carbapenem and cephalosporin stress. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-021-02136-y. BioMed Central 2021-03-09 /pmc/articles/PMC7941987/ /pubmed/33750290 http://dx.doi.org/10.1186/s12866-021-02136-y Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, 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 changes were made. 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/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Baishya, Somorita
Deshamukhya, Chandrayee
Wangkheimayum, Jayalaxmi
Das, Bhaskar Jyoti
Anbarasu, Anand
Talukdar, Anupam Das
Bhattacharjee, Amitabha
Choudhury, Manabendra Dutta
Transcriptional expression of secondary resistance genes ccdB and repA2 is enhanced in presence of cephalosporin and carbapenem in Escherichia coli
title Transcriptional expression of secondary resistance genes ccdB and repA2 is enhanced in presence of cephalosporin and carbapenem in Escherichia coli
title_full Transcriptional expression of secondary resistance genes ccdB and repA2 is enhanced in presence of cephalosporin and carbapenem in Escherichia coli
title_fullStr Transcriptional expression of secondary resistance genes ccdB and repA2 is enhanced in presence of cephalosporin and carbapenem in Escherichia coli
title_full_unstemmed Transcriptional expression of secondary resistance genes ccdB and repA2 is enhanced in presence of cephalosporin and carbapenem in Escherichia coli
title_short Transcriptional expression of secondary resistance genes ccdB and repA2 is enhanced in presence of cephalosporin and carbapenem in Escherichia coli
title_sort transcriptional expression of secondary resistance genes ccdb and repa2 is enhanced in presence of cephalosporin and carbapenem in escherichia coli
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7941987/
https://www.ncbi.nlm.nih.gov/pubmed/33750290
http://dx.doi.org/10.1186/s12866-021-02136-y
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