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Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring bla(VIM-2), bla(IMP-1) and bla(OXA-23) Genes by Using Loop-Mediated Isothermal Amplification Methods

BACKGROUND: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) and Acinetobacter baumannii (CRAB) are the leading causes of nosocomial infections. A rapid and sensitive test to detect CRPA and CRAB is required for appropriate antibiotic treatment. We optimized a loop-mediated isothermal amplificatio...

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Autores principales: Kim, Hye Jin, Kim, Hyung Sun, Lee, Jae Myun, Yoon, Sang Sun, Yong, Dongeun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Korean Society for Laboratory Medicine 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4697338/
https://www.ncbi.nlm.nih.gov/pubmed/26522754
http://dx.doi.org/10.3343/alm.2016.36.1.15
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author Kim, Hye Jin
Kim, Hyung Sun
Lee, Jae Myun
Yoon, Sang Sun
Yong, Dongeun
author_facet Kim, Hye Jin
Kim, Hyung Sun
Lee, Jae Myun
Yoon, Sang Sun
Yong, Dongeun
author_sort Kim, Hye Jin
collection PubMed
description BACKGROUND: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) and Acinetobacter baumannii (CRAB) are the leading causes of nosocomial infections. A rapid and sensitive test to detect CRPA and CRAB is required for appropriate antibiotic treatment. We optimized a loop-mediated isothermal amplification (LAMP) assay to detect the presence of bla(VIM-2), bla(IMP-1), and bla(OXA-23), which are critical components for carbapenem resistance. METHODS: Two sets of primers, inner and outer primers, were manually designed as previously described. The LAMP buffer was optimized (at 2mM MgSO(4)) by testing different concentrations of MgSO(4). The optimal reaction temperature and incubation time were determined by using a gradient thermocycler. Then, the optimized bla(VIM-2), bla(IMP-1), and bla(OXA-23) LAMP reactions were evaluated by using 120 P. aeruginosa and 99 A. baumannii clinical isolates. RESULTS: Only one strain of the 100 CRPA isolates harbored bla(IMP-1), whereas none of them harbored bla(VIM-2). These results indicate that the acquisition of bla(VIM-2) or bla(IMP-1) may not play a major role in carbapenem resistance in Korea. Fifty two strains of the 75 CRAB isolates contained bla(OXA-23), but none contained bla(VIM-2) and bla(IMP-1) alleles. CONCLUSIONS: Our results demonstrate the usefulness of LAMP for the diagnosis of CRPA and CRAB.
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spelling pubmed-46973382016-01-04 Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring bla(VIM-2), bla(IMP-1) and bla(OXA-23) Genes by Using Loop-Mediated Isothermal Amplification Methods Kim, Hye Jin Kim, Hyung Sun Lee, Jae Myun Yoon, Sang Sun Yong, Dongeun Ann Lab Med Original Article BACKGROUND: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) and Acinetobacter baumannii (CRAB) are the leading causes of nosocomial infections. A rapid and sensitive test to detect CRPA and CRAB is required for appropriate antibiotic treatment. We optimized a loop-mediated isothermal amplification (LAMP) assay to detect the presence of bla(VIM-2), bla(IMP-1), and bla(OXA-23), which are critical components for carbapenem resistance. METHODS: Two sets of primers, inner and outer primers, were manually designed as previously described. The LAMP buffer was optimized (at 2mM MgSO(4)) by testing different concentrations of MgSO(4). The optimal reaction temperature and incubation time were determined by using a gradient thermocycler. Then, the optimized bla(VIM-2), bla(IMP-1), and bla(OXA-23) LAMP reactions were evaluated by using 120 P. aeruginosa and 99 A. baumannii clinical isolates. RESULTS: Only one strain of the 100 CRPA isolates harbored bla(IMP-1), whereas none of them harbored bla(VIM-2). These results indicate that the acquisition of bla(VIM-2) or bla(IMP-1) may not play a major role in carbapenem resistance in Korea. Fifty two strains of the 75 CRAB isolates contained bla(OXA-23), but none contained bla(VIM-2) and bla(IMP-1) alleles. CONCLUSIONS: Our results demonstrate the usefulness of LAMP for the diagnosis of CRPA and CRAB. The Korean Society for Laboratory Medicine 2016-01 2015-10-26 /pmc/articles/PMC4697338/ /pubmed/26522754 http://dx.doi.org/10.3343/alm.2016.36.1.15 Text en © The Korean Society for Laboratory Medicine. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Kim, Hye Jin
Kim, Hyung Sun
Lee, Jae Myun
Yoon, Sang Sun
Yong, Dongeun
Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring bla(VIM-2), bla(IMP-1) and bla(OXA-23) Genes by Using Loop-Mediated Isothermal Amplification Methods
title Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring bla(VIM-2), bla(IMP-1) and bla(OXA-23) Genes by Using Loop-Mediated Isothermal Amplification Methods
title_full Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring bla(VIM-2), bla(IMP-1) and bla(OXA-23) Genes by Using Loop-Mediated Isothermal Amplification Methods
title_fullStr Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring bla(VIM-2), bla(IMP-1) and bla(OXA-23) Genes by Using Loop-Mediated Isothermal Amplification Methods
title_full_unstemmed Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring bla(VIM-2), bla(IMP-1) and bla(OXA-23) Genes by Using Loop-Mediated Isothermal Amplification Methods
title_short Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring bla(VIM-2), bla(IMP-1) and bla(OXA-23) Genes by Using Loop-Mediated Isothermal Amplification Methods
title_sort rapid detection of pseudomonas aeruginosa and acinetobacter baumannii harboring bla(vim-2), bla(imp-1) and bla(oxa-23) genes by using loop-mediated isothermal amplification methods
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4697338/
https://www.ncbi.nlm.nih.gov/pubmed/26522754
http://dx.doi.org/10.3343/alm.2016.36.1.15
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