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Title
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Evaluation of a loop-mediated isothermal amplification assay to detect carbapenemases directly from bronchoalveolar lavage fluid spiked with Acinetobacter spp.
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Author
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Abstract
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| Carbapenem-resistant Acinetobacter spp. mainly Acinetobacter baumannii are frequently causing nosocomial infections with high mortality. In this study, the efficacy of the Eazyplex(R) 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 (bla(KPC), bla(NDM), bla(VIM), bla(OXA-48), bla(OXA-23), bla(OXA-40), and bla(OXA-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 10(2) and 10(3) 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 10(2) CFU/ml; therefore, the limit of sensitivity is 10(3) 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 10(3) CFU/ml. |
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Language
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English
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Source (journal)
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Frontiers in microbiology. - Lausanne, 2010, currens
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Publication
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Lausanne
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Frontiers Research Foundation
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2021
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ISSN
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1664-302X
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DOI
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10.3389/FMICB.2020.597684
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Volume/pages
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11
(2021)
, 4 p.
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Article Reference
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597684
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ISI
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000612073400001
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Pubmed ID
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33519735
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Medium
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E-only publicatie
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Full text (Publisher's DOI)
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Full text (open access)
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