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A highly multiplexed melt-curve assay for detecting the most prevalent carbapenemase, ESBL and AmpC genes


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Edwards, Thomas, Williams, Chris, Sealey, J, Sasaki, S, Hobbs, G, Cuevas, Luis ORCID:, Evans, K and Adams, Emily ORCID: (2020) 'A highly multiplexed melt-curve assay for detecting the most prevalent carbapenemase, ESBL and AmpC genes'. Diagnostic Microbiology and Infectious Disease, Vol 97, Issue 4, p. 115076.

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Resistance to third generation cephalosporins and carbapenems in Gram-negative bacteria is chiefly mediated by beta-lactamases including ESBL, AmpC and carbapenemase enzymes. Routine phenotypic detection methods do not provide timely results, and there is a lack of comprehensive molecular panels covering all important markers.
An ESBL/carbapenemase HRM assay (SHV, TEM, CTX-M ESBL families, and NDM, IMP, KPC, VIM and OXA-48-like carbapenemases) and an AmpC HRM assay (16S rDNA control, FOX, MOX, ACC, EBC, CIT and DHA) were designed, and evaluated on 111 Gram-negative isolates with mixed resistance patterns.
The sensitivity for carbapenemase, ESBL and AmpC genes was 96.7% (95%CI:82.8-99.9%), 93.6% (95%CI:85.7-97.9%) and 93.8% (95%CI:82.8-98.7%), respectively with a specificity of 100% (95%CI:95.6-100%), 93.9% (95%CI:79.8-99.3%) and 93.7% (95%CI:84.5-98.2%).
The HRM assays enable the simultaneous detection of the fourteen most important ESBL, carbapenemase and AmpC genes and could be used as a molecular surveillance tool or to hasten detection of AMR for treatment management.

Item Type: Article
Subjects: QU Biochemistry > Enzymes > QU 136 Hydrolases
QV Pharmacology > Anti-Bacterial Agents. Tissue Extracts > QV 350.5.C3 Cephalosporins
QW Microbiology and Immunology > Bacteria > QW 131 Gram-negative bacteria.
QW Microbiology and Immunology > QW 45 Microbial drug resistance. General or not elsewhere classified.
WB Practice of Medicine > Therapeutics > WB 330 Drug therapy
Faculty: Department: Biological Sciences > Department of Tropical Disease Biology
Clinical Sciences & International Health > Clinical Sciences Department
Digital Object Identifer (DOI):
Depositing User: Claire McIntyre
Date Deposited: 14 May 2020 12:37
Last Modified: 12 Aug 2020 09:59


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