A novel air-dried multiplex high-resolution melt assay for the detection of extended-spectrum β-lactamase and carbapenemase genes.
Antimicrobial resistance
Carbapenemase
ESBL
Extended-spectrum β-lactamase
High-resolution melting
Molecular diagnostics
Journal
Journal of global antimicrobial resistance
ISSN: 2213-7173
Titre abrégé: J Glob Antimicrob Resist
Pays: Netherlands
ID NLM: 101622459
Informations de publication
Date de publication:
12 2021
12 2021
Historique:
received:
28
06
2021
revised:
11
08
2021
accepted:
19
08
2021
pubmed:
6
9
2021
medline:
31
12
2021
entrez:
5
9
2021
Statut:
ppublish
Résumé
This study aimed to develop and evaluate a novel air-dried high-resolution melt (HRM) assay to detect eight major extended-spectrum β-lactamase (ESBL) (bla The assay was evaluated using 439 DNA samples extracted from bacterial isolates from Nepal, Malawi and the UK and 390 clinical isolates from Nepal with known antimicrobial susceptibility. Assay reproducibility was evaluated across five different real-time quantitative PCR (qPCR) instruments [Rotor-Gene® Q, QuantStudio The sensitivity and specificity (with 95% confidence intervals) for detecting ESBL and carbapenemase genes was 94.7% (92.5-96.5%) and 99.2% (98.8-99.5%) compared with the reference gel-based PCR and sequencing and 98.3% (97.0-99.3%) and 98.5% (98.0-98.9%) compared with the original HRM wet PCR mix format. Overall agreement was 91.1% (90.0-92.9%) when predicting phenotypic resistance to cefotaxime and meropenem among Enterobacteriaceae isolates. We observed almost perfect inter-machine reproducibility of the air-dried HRM assay, and no loss of sensitivity occurred under all storage conditions and time points. We present a ready-to-use air-dried HRM PCR assay that offers an easy, thermostable, fast and accurate tool for the detection of ESBL and carbapenemase genes in DNA samples to improve antimicrobial resistance detection.
Identifiants
pubmed: 34482019
pii: S2213-7165(21)00197-1
doi: 10.1016/j.jgar.2021.08.006
pmc: PMC8692233
pii:
doi:
Substances chimiques
Anti-Bacterial Agents
0
Bacterial Proteins
0
beta-Lactamases
EC 3.5.2.6
carbapenemase
EC 3.5.2.6
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
123-131Subventions
Organisme : Medical Research Council
ID : 17196
Pays : United Kingdom
Informations de copyright
Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.
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