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Title
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A comparative study between real-time PCR and loop-mediated isothermal amplification to detect carbapenemase and/or ESBL genes in Enterobacteriaceae directly from bronchoalveolar lavage fluid samples
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Author
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Abstract
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| Objectives: To evaluate and compare the efficacy of real-time PCR (Xpert Carba-R) and loop-mediated isothermal amplification (Eazyplex (R) SuperBug CRE) for detecting carbapenemase carriage in Enterobacteriaceae directly from bronchoalveolar lavage (BAL). Methods: Negative BAL samples were spiked with 21 well-characterized carbapenemase-producing Enterobacteriaceae strains to a final concentration of 10(2)-10(4) cfu/mL. Xpert Carba-R (Cepheid, Sunnyvale, CA, USA), which detects five targets (bla(KPC), bla(NDM), bla(VIM), bla(OXA-48) and bla(IMP-1)), and the Eazyplex (R) SuperBug CRE system (Amplex-Diagnostics GmbH, Germany), which detects seven genes (bla(KPC), bla(NDM), bla(VIM), bla(OXA-48), bla(OXA-181), bla(CTXM-1) and bla(CTXM-9)), were evaluated for the detection of these genes directly from BAL samples. Results: Xpert Carba-R showed 100% agreement with carbapenemase characterization by PCR and sequencing for all final bacteria concentrations. Eazyplex (R) SuperBug CRE showed 100%, 80% and 27% agreement with PCR and sequencing when testing 10(4), 10(3) and 10(2) cfu/mL, respectively. False negative results for Eazyplex (R) SuperBug CRE matched the highest cycle threshold values for Xpert Carba-R. Hands-on time for both assays was about 15 min, but Eazyplex (R) SuperBug CRE results were available within 30 min, whereas Xpert Carba-R took around 50 min. Conclusions: We here describe the successful use of two commercial diagnostic tests, Xpert Carba-R and Eazyplex (R) SuperBug CRE, to detect bacterial carbapenem resistance genes directly in lower respiratory tract samples. Our results could be used as proof-of-concept data for validation of these tests for this indication. |
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Language
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English
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Source (journal)
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The journal of antimicrobial chemotherapy. - London, 1975, currens
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Publication
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London
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2020
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ISSN
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0305-7453
[print]
1460-2091
[online]
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DOI
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10.1093/JAC/DKAA031
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Volume/pages
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75
:6
(2020)
, p. 1453-1457
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ISI
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000567185800013
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Pubmed ID
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32073602
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Full text (Publisher's DOI)
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Full text (publisher's version - intranet only)
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