Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter spp

Carbapenem-resistant Acinetobacter spp. mainly Acinetobacter baumannii are frequently causing nosocomial infections with high mortality. In this study, the efficacy of the Eazyplex® SuperBug Complete A system, based on loop-mediated isothermal amplification (LAMP), to detect the presence of carbapen...

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Autores: Moreno-Morales, Javier, Vergara, Andrea, Kostyanev, Tomislav, Rodríguez-Baño, Jesús, Goossens, Herman, Vila, Jordi
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Universidad de Sevilla (US)
Repositorio:idUS. Depósito de Investigación de la Universidad de Sevilla
OAI Identifier:oai:idus.us.es:11441/137234
Acceso en línea:https://hdl.handle.net/11441/137234
https://doi.org/10.3389/fmicb.2020.597684
Access Level:acceso abierto
Palabra clave:Carbapenemases
Acinetobacter spp.
Bronchoalveolar lavage
Detection
Oxacillinases
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spelling Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter sppMoreno-Morales, JavierVergara, AndreaKostyanev, TomislavRodríguez-Baño, JesúsGoossens, HermanVila, JordiCarbapenemasesAcinetobacter spp.Bronchoalveolar lavageDetectionOxacillinasesCarbapenem-resistant Acinetobacter spp. mainly Acinetobacter baumannii are frequently causing nosocomial infections with high mortality. In this study, the efficacy of the Eazyplex® SuperBug Complete A system, based on loop-mediated isothermal amplification (LAMP), to detect the presence of carbapenemases in Acinetobacter spp. directly from bronchoalveolar lavage (BAL) samples was assessed. A total of 22 Acinetobacter spp. strains producing OXA-23, OXA-40, OXA-58, NDM, and IMP were selected. Eazyplex SuperBug Complete A kit, used with the Genie II device, is a molecular diagnostics kit that detects a selection of genes that express carbapenemases (blaKPC, blaNDM, blaVIM, blaOXA–48, blaOXA–23, blaOXA–40, and blaOXA–58). Negative BAL samples were identified, McFarland solutions were prepared from each of the 22 Acinetobacter strains and serial dilutions in saline solution were made to finally spike BAL samples to a concentration of 102 and 103 CFU/ml. Fifteen concentrations out of the 44 tested out did not provide detection of the carbapenemase-producing gene, all but one being at the lowest concentration tested at 102 CFU/ml; therefore, the limit of sensitivity is 103 CFU/ml. This assay represents the kind of advantages that investing in molecular diagnostics brings to the clinical practice, allowing the identification of carbapenemases in less than 30 min with a sensitivity of 103 CFU/ml.Spanish Ministry of Science and Innovation through the “Centro de Excelencia Severo Ochoa"Generalitat de CatalunyaMethodsMedicina2021info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttps://hdl.handle.net/11441/137234https://doi.org/10.3389/fmicb.2020.597684reponame:idUS. Depósito de Investigación de la Universidad de Sevillainstname:Universidad de Sevilla (US)InglésFrontiers in microbiology, 11CEX2018-000806-SCERCA Programhttps://www.frontiersin.org/articles/10.3389/fmicb.2020.597684/fullinfo:eu-repo/semantics/openAccessoai:idus.us.es:11441/1372342026-06-17T12:51:07Z
dc.title.none.fl_str_mv Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter spp
title Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter spp
spellingShingle Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter spp
Moreno-Morales, Javier
Carbapenemases
Acinetobacter spp.
Bronchoalveolar lavage
Detection
Oxacillinases
title_short Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter spp
title_full Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter spp
title_fullStr Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter spp
title_full_unstemmed Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter spp
title_sort Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with acinetobacter spp
dc.creator.none.fl_str_mv Moreno-Morales, Javier
Vergara, Andrea
Kostyanev, Tomislav
Rodríguez-Baño, Jesús
Goossens, Herman
Vila, Jordi
author Moreno-Morales, Javier
author_facet Moreno-Morales, Javier
Vergara, Andrea
Kostyanev, Tomislav
Rodríguez-Baño, Jesús
Goossens, Herman
Vila, Jordi
author_role author
author2 Vergara, Andrea
Kostyanev, Tomislav
Rodríguez-Baño, Jesús
Goossens, Herman
Vila, Jordi
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Medicina
dc.subject.none.fl_str_mv Carbapenemases
Acinetobacter spp.
Bronchoalveolar lavage
Detection
Oxacillinases
topic Carbapenemases
Acinetobacter spp.
Bronchoalveolar lavage
Detection
Oxacillinases
description Carbapenem-resistant Acinetobacter spp. mainly Acinetobacter baumannii are frequently causing nosocomial infections with high mortality. In this study, the efficacy of the Eazyplex® SuperBug Complete A system, based on loop-mediated isothermal amplification (LAMP), to detect the presence of carbapenemases in Acinetobacter spp. directly from bronchoalveolar lavage (BAL) samples was assessed. A total of 22 Acinetobacter spp. strains producing OXA-23, OXA-40, OXA-58, NDM, and IMP were selected. Eazyplex SuperBug Complete A kit, used with the Genie II device, is a molecular diagnostics kit that detects a selection of genes that express carbapenemases (blaKPC, blaNDM, blaVIM, blaOXA–48, blaOXA–23, blaOXA–40, and blaOXA–58). Negative BAL samples were identified, McFarland solutions were prepared from each of the 22 Acinetobacter strains and serial dilutions in saline solution were made to finally spike BAL samples to a concentration of 102 and 103 CFU/ml. Fifteen concentrations out of the 44 tested out did not provide detection of the carbapenemase-producing gene, all but one being at the lowest concentration tested at 102 CFU/ml; therefore, the limit of sensitivity is 103 CFU/ml. This assay represents the kind of advantages that investing in molecular diagnostics brings to the clinical practice, allowing the identification of carbapenemases in less than 30 min with a sensitivity of 103 CFU/ml.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/11441/137234
https://doi.org/10.3389/fmicb.2020.597684
url https://hdl.handle.net/11441/137234
https://doi.org/10.3389/fmicb.2020.597684
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Frontiers in microbiology, 11
CEX2018-000806-S
CERCA Program
https://www.frontiersin.org/articles/10.3389/fmicb.2020.597684/full
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Methods
publisher.none.fl_str_mv Methods
dc.source.none.fl_str_mv reponame:idUS. Depósito de Investigación de la Universidad de Sevilla
instname:Universidad de Sevilla (US)
instname_str Universidad de Sevilla (US)
reponame_str idUS. Depósito de Investigación de la Universidad de Sevilla
collection idUS. Depósito de Investigación de la Universidad de Sevilla
repository.name.fl_str_mv
repository.mail.fl_str_mv
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