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Recombinase Polymerase Amplification Combined with Lateral Flow Strip for Rapid Detection of OXA-48-like Carbapenemase Genes in Enterobacterales

Carbapenem-resistant Enterobacterales (CRE) possessing various carbapenemases, particularly the OXA-48 group, are now rapidly spreading and becoming a major public health concern worldwide. Phenotypic detection of OXA-48-like carbapenemases is still suboptimal due to their weak carbapenemase activit...

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Autores principales: Hemwaranon, Phatsarawadee, Srisrattakarn, Arpasiri, Lulitanond, Aroonlug, Tippayawat, Patcharaporn, Tavichakorntrakool, Ratree, Wonglakorn, Lumyai, Daduang, Jureerut, Chanawong, Aroonwadee
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9686806/
https://www.ncbi.nlm.nih.gov/pubmed/36358154
http://dx.doi.org/10.3390/antibiotics11111499
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author Hemwaranon, Phatsarawadee
Srisrattakarn, Arpasiri
Lulitanond, Aroonlug
Tippayawat, Patcharaporn
Tavichakorntrakool, Ratree
Wonglakorn, Lumyai
Daduang, Jureerut
Chanawong, Aroonwadee
author_facet Hemwaranon, Phatsarawadee
Srisrattakarn, Arpasiri
Lulitanond, Aroonlug
Tippayawat, Patcharaporn
Tavichakorntrakool, Ratree
Wonglakorn, Lumyai
Daduang, Jureerut
Chanawong, Aroonwadee
author_sort Hemwaranon, Phatsarawadee
collection PubMed
description Carbapenem-resistant Enterobacterales (CRE) possessing various carbapenemases, particularly the OXA-48 group, are now rapidly spreading and becoming a major public health concern worldwide. Phenotypic detection of OXA-48-like carbapenemases is still suboptimal due to their weak carbapenemase activity, whereas highly sensitive and specific polymerase chain reaction (PCR)-based methods take at least 3–4 h. We, therefore, developed a recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip assay for the rapid detection of bla(OXA-48-like) in Enterobacterales. A total of 131 clinical isolates including 61 bla(OXA-48-like)-carrying Enterobacterales isolates and 70 Gram-negative bacilli isolates containing other bla genes were subjected to the RPA method performed under isothermal conditions at 37 °C within 10 min and visually inspected by LF strip within 5 min. The RPA-LF assay provided 100% sensitivity (95% confidence interval, 92.6–100%) and 100% specificity (93.5–100%) for detecting bla(OXA-48-like) genes from bacterial colonies. Its detection limit was 100 times less than that of the PCR method. This assay is rapid, easy to perform, and provides excellent performance without any special equipment. It may be applied for directly identifying the bla(OXA-48-like) genes in Enterobacterales obtained from blood culture. Rapid identification of carbapenemase types is essential for selecting appropriate antimicrobial options, particularly the β-lactams combined with novel β-lactamase inhibitors.
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spelling pubmed-96868062022-11-25 Recombinase Polymerase Amplification Combined with Lateral Flow Strip for Rapid Detection of OXA-48-like Carbapenemase Genes in Enterobacterales Hemwaranon, Phatsarawadee Srisrattakarn, Arpasiri Lulitanond, Aroonlug Tippayawat, Patcharaporn Tavichakorntrakool, Ratree Wonglakorn, Lumyai Daduang, Jureerut Chanawong, Aroonwadee Antibiotics (Basel) Article Carbapenem-resistant Enterobacterales (CRE) possessing various carbapenemases, particularly the OXA-48 group, are now rapidly spreading and becoming a major public health concern worldwide. Phenotypic detection of OXA-48-like carbapenemases is still suboptimal due to their weak carbapenemase activity, whereas highly sensitive and specific polymerase chain reaction (PCR)-based methods take at least 3–4 h. We, therefore, developed a recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip assay for the rapid detection of bla(OXA-48-like) in Enterobacterales. A total of 131 clinical isolates including 61 bla(OXA-48-like)-carrying Enterobacterales isolates and 70 Gram-negative bacilli isolates containing other bla genes were subjected to the RPA method performed under isothermal conditions at 37 °C within 10 min and visually inspected by LF strip within 5 min. The RPA-LF assay provided 100% sensitivity (95% confidence interval, 92.6–100%) and 100% specificity (93.5–100%) for detecting bla(OXA-48-like) genes from bacterial colonies. Its detection limit was 100 times less than that of the PCR method. This assay is rapid, easy to perform, and provides excellent performance without any special equipment. It may be applied for directly identifying the bla(OXA-48-like) genes in Enterobacterales obtained from blood culture. Rapid identification of carbapenemase types is essential for selecting appropriate antimicrobial options, particularly the β-lactams combined with novel β-lactamase inhibitors. MDPI 2022-10-28 /pmc/articles/PMC9686806/ /pubmed/36358154 http://dx.doi.org/10.3390/antibiotics11111499 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hemwaranon, Phatsarawadee
Srisrattakarn, Arpasiri
Lulitanond, Aroonlug
Tippayawat, Patcharaporn
Tavichakorntrakool, Ratree
Wonglakorn, Lumyai
Daduang, Jureerut
Chanawong, Aroonwadee
Recombinase Polymerase Amplification Combined with Lateral Flow Strip for Rapid Detection of OXA-48-like Carbapenemase Genes in Enterobacterales
title Recombinase Polymerase Amplification Combined with Lateral Flow Strip for Rapid Detection of OXA-48-like Carbapenemase Genes in Enterobacterales
title_full Recombinase Polymerase Amplification Combined with Lateral Flow Strip for Rapid Detection of OXA-48-like Carbapenemase Genes in Enterobacterales
title_fullStr Recombinase Polymerase Amplification Combined with Lateral Flow Strip for Rapid Detection of OXA-48-like Carbapenemase Genes in Enterobacterales
title_full_unstemmed Recombinase Polymerase Amplification Combined with Lateral Flow Strip for Rapid Detection of OXA-48-like Carbapenemase Genes in Enterobacterales
title_short Recombinase Polymerase Amplification Combined with Lateral Flow Strip for Rapid Detection of OXA-48-like Carbapenemase Genes in Enterobacterales
title_sort recombinase polymerase amplification combined with lateral flow strip for rapid detection of oxa-48-like carbapenemase genes in enterobacterales
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9686806/
https://www.ncbi.nlm.nih.gov/pubmed/36358154
http://dx.doi.org/10.3390/antibiotics11111499
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