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Rapid Detection of Enterobacter cloacae With a Visualized Isothermal Recombinase Polymerase Amplification Assay

Enterobacter cloacae exhibits strong adhesion and invasion properties that contribute its ability to infect the host; it is considered an important opportunistic pathogen throughout the world. To control the spread of E. cloacae, simple, rapid, and accurate detection methods are required. Current me...

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Autores principales: Hu, Juan, Xu, Jing, Lu, Yingzhi, Wang, Lei, Wang, Yan, Chen, Cheng, Zhu, Wenjun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240108/
https://www.ncbi.nlm.nih.gov/pubmed/37273073
http://dx.doi.org/10.1007/s00284-023-03269-1
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author Hu, Juan
Xu, Jing
Lu, Yingzhi
Wang, Lei
Wang, Yan
Chen, Cheng
Zhu, Wenjun
author_facet Hu, Juan
Xu, Jing
Lu, Yingzhi
Wang, Lei
Wang, Yan
Chen, Cheng
Zhu, Wenjun
author_sort Hu, Juan
collection PubMed
description Enterobacter cloacae exhibits strong adhesion and invasion properties that contribute its ability to infect the host; it is considered an important opportunistic pathogen throughout the world. To control the spread of E. cloacae, simple, rapid, and accurate detection methods are required. Current methods suffer from various shortcomings and do not meet the demand for on-site quickly detection. Using recombinase polymerase amplification combined with lateral flow strip (RPA-LFS), an isothermal detection method was developed to target the outer membrane protein X (ompX) gene of E. cloacae. This reaction can be performed in 30 min at 37 °C. Limit of detection of 10 CFU/reaction was equivalent to that of the qPCR method. The detection accuracy of clinical samples was also equal to that of the qPCR method. In this study, we developed the RPA-LFS assay, which is simple, rapid, accurate, and does not require a laboratory facility. This assay may prove useful for detecting E. cloacae on-site.
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spelling pubmed-102401082023-06-06 Rapid Detection of Enterobacter cloacae With a Visualized Isothermal Recombinase Polymerase Amplification Assay Hu, Juan Xu, Jing Lu, Yingzhi Wang, Lei Wang, Yan Chen, Cheng Zhu, Wenjun Curr Microbiol Article Enterobacter cloacae exhibits strong adhesion and invasion properties that contribute its ability to infect the host; it is considered an important opportunistic pathogen throughout the world. To control the spread of E. cloacae, simple, rapid, and accurate detection methods are required. Current methods suffer from various shortcomings and do not meet the demand for on-site quickly detection. Using recombinase polymerase amplification combined with lateral flow strip (RPA-LFS), an isothermal detection method was developed to target the outer membrane protein X (ompX) gene of E. cloacae. This reaction can be performed in 30 min at 37 °C. Limit of detection of 10 CFU/reaction was equivalent to that of the qPCR method. The detection accuracy of clinical samples was also equal to that of the qPCR method. In this study, we developed the RPA-LFS assay, which is simple, rapid, accurate, and does not require a laboratory facility. This assay may prove useful for detecting E. cloacae on-site. Springer US 2023-06-05 2023 /pmc/articles/PMC10240108/ /pubmed/37273073 http://dx.doi.org/10.1007/s00284-023-03269-1 Text en © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Article
Hu, Juan
Xu, Jing
Lu, Yingzhi
Wang, Lei
Wang, Yan
Chen, Cheng
Zhu, Wenjun
Rapid Detection of Enterobacter cloacae With a Visualized Isothermal Recombinase Polymerase Amplification Assay
title Rapid Detection of Enterobacter cloacae With a Visualized Isothermal Recombinase Polymerase Amplification Assay
title_full Rapid Detection of Enterobacter cloacae With a Visualized Isothermal Recombinase Polymerase Amplification Assay
title_fullStr Rapid Detection of Enterobacter cloacae With a Visualized Isothermal Recombinase Polymerase Amplification Assay
title_full_unstemmed Rapid Detection of Enterobacter cloacae With a Visualized Isothermal Recombinase Polymerase Amplification Assay
title_short Rapid Detection of Enterobacter cloacae With a Visualized Isothermal Recombinase Polymerase Amplification Assay
title_sort rapid detection of enterobacter cloacae with a visualized isothermal recombinase polymerase amplification assay
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240108/
https://www.ncbi.nlm.nih.gov/pubmed/37273073
http://dx.doi.org/10.1007/s00284-023-03269-1
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