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Rapid optical determination of β-lactamase and antibiotic activity

BACKGROUND: The absence of rapid tests evaluating antibiotic susceptibility results in the empirical prescription of antibiotics. This can lead to treatment failures due to escalating antibiotic resistance, and also furthers the emergence of drug-resistant bacteria. This study reports a rapid optica...

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Autores principales: Khan, Shazia, Sallum, Ulysses W, Zheng, Xiang, Nau, Gerard J, Hasan, Tayyaba
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4234275/
https://www.ncbi.nlm.nih.gov/pubmed/24708478
http://dx.doi.org/10.1186/1471-2180-14-84
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author Khan, Shazia
Sallum, Ulysses W
Zheng, Xiang
Nau, Gerard J
Hasan, Tayyaba
author_facet Khan, Shazia
Sallum, Ulysses W
Zheng, Xiang
Nau, Gerard J
Hasan, Tayyaba
author_sort Khan, Shazia
collection PubMed
description BACKGROUND: The absence of rapid tests evaluating antibiotic susceptibility results in the empirical prescription of antibiotics. This can lead to treatment failures due to escalating antibiotic resistance, and also furthers the emergence of drug-resistant bacteria. This study reports a rapid optical method to detect β-lactamase and thereby assess activity of β-lactam antibiotics, which could provide an approach for targeted prescription of antibiotics. The methodology is centred on a fluorescence quenching based probe (β-LEAF – β-Lactamase Enzyme Activated Fluorophore) that mimics the structure of β-lactam antibiotics. RESULTS: The β-LEAF assay was performed for rapid determination of β-lactamase production and activity of β-lactam antibiotic (cefazolin) on a panel of Staphylococcus aureus ATCC strains and clinical isolates. Four of the clinical isolates were determined to be lactamase producers, with the capacity to inactivate cefazolin, out of the twenty-five isolates tested. These results were compared against gold standard methods, nitrocefin disk test for β-lactamase detection and disk diffusion for antibiotic susceptibility, showing results to be largely consistent. Furthermore, in the sub-set of β-lactamase producers, it was demonstrated and validated that multiple antibiotics (cefazolin, cefoxitin, cefepime) could be assessed simultaneously to predict the antibiotic that would be most active for a given bacterial isolate. CONCLUSIONS: The study establishes the rapid β-LEAF assay for β-lactamase detection and prediction of antibiotic activity using S. aureus clinical isolates. Although the focus in the current study is β-lactamase-based resistance, the overall approach represents a broad diagnostic platform. In the long-term, these studies form the basis for the development of assays utilizing a broader variety of targets, pathogens and drugs.
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spelling pubmed-42342752014-11-18 Rapid optical determination of β-lactamase and antibiotic activity Khan, Shazia Sallum, Ulysses W Zheng, Xiang Nau, Gerard J Hasan, Tayyaba BMC Microbiol Research Article BACKGROUND: The absence of rapid tests evaluating antibiotic susceptibility results in the empirical prescription of antibiotics. This can lead to treatment failures due to escalating antibiotic resistance, and also furthers the emergence of drug-resistant bacteria. This study reports a rapid optical method to detect β-lactamase and thereby assess activity of β-lactam antibiotics, which could provide an approach for targeted prescription of antibiotics. The methodology is centred on a fluorescence quenching based probe (β-LEAF – β-Lactamase Enzyme Activated Fluorophore) that mimics the structure of β-lactam antibiotics. RESULTS: The β-LEAF assay was performed for rapid determination of β-lactamase production and activity of β-lactam antibiotic (cefazolin) on a panel of Staphylococcus aureus ATCC strains and clinical isolates. Four of the clinical isolates were determined to be lactamase producers, with the capacity to inactivate cefazolin, out of the twenty-five isolates tested. These results were compared against gold standard methods, nitrocefin disk test for β-lactamase detection and disk diffusion for antibiotic susceptibility, showing results to be largely consistent. Furthermore, in the sub-set of β-lactamase producers, it was demonstrated and validated that multiple antibiotics (cefazolin, cefoxitin, cefepime) could be assessed simultaneously to predict the antibiotic that would be most active for a given bacterial isolate. CONCLUSIONS: The study establishes the rapid β-LEAF assay for β-lactamase detection and prediction of antibiotic activity using S. aureus clinical isolates. Although the focus in the current study is β-lactamase-based resistance, the overall approach represents a broad diagnostic platform. In the long-term, these studies form the basis for the development of assays utilizing a broader variety of targets, pathogens and drugs. BioMed Central 2014-04-04 /pmc/articles/PMC4234275/ /pubmed/24708478 http://dx.doi.org/10.1186/1471-2180-14-84 Text en Copyright © 2014 Khan et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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.
spellingShingle Research Article
Khan, Shazia
Sallum, Ulysses W
Zheng, Xiang
Nau, Gerard J
Hasan, Tayyaba
Rapid optical determination of β-lactamase and antibiotic activity
title Rapid optical determination of β-lactamase and antibiotic activity
title_full Rapid optical determination of β-lactamase and antibiotic activity
title_fullStr Rapid optical determination of β-lactamase and antibiotic activity
title_full_unstemmed Rapid optical determination of β-lactamase and antibiotic activity
title_short Rapid optical determination of β-lactamase and antibiotic activity
title_sort rapid optical determination of β-lactamase and antibiotic activity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4234275/
https://www.ncbi.nlm.nih.gov/pubmed/24708478
http://dx.doi.org/10.1186/1471-2180-14-84
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