title: Rapid and accurate detection of carbapenemase genes in Enterobacteriaceae with the Cepheid Xpert Carba-R assay.
abstract: Rapid and accurate detection of carbapenemase-producing Enterobacteriaceae is pivotal for adequate antibiotic therapy and infection control. The Cepheid Xpert Carba-R assay detects and identifies the most prevalent carbapenemases (KPC, VIM, IMP, NDM and OXA-48), using automated real-time PCR. The test performance of the Xpert Carba-R was evaluated with 129 well-characterized non-repeat Enterobacteriaceae isolates, suspected for carbapenemase production, i.e. with meropenem MICs >0.25 mg l-1. The isolate collection contained 100 carbapenemase-producing isolates (36 KPC-2 or KPC-3, 20 VIM-1, 4 KPC-2 plus VIM-1, 5 NDM-1, 2 IMP-1, 1 IMP-28, 1 IMP-1 plus VIM-1 and 31 OXA-48 like) and 29 negative control isolates producing extended-spectrum β-lactamase and/or AmpC β-lactamases. PCR and sequencing of β-lactamase genes were used as reference tests. The sensitivity of the Xpert Carba-R was 100 % (100/100), with a 100 % (29/29) specificity. The time to result was approximately 55 min with a hands-on time of only 1 min per isolate. In conclusion, the Carba-R assay is a rapid and accurate instrument for the confirmation and identification of the blaKPC, blaVIM, blaIMP, blaNDM and blaOXA-48 genes.
authors.affiliation: Department of Medical Microbiology, Medisch Centrum Alkmaar, Juliana Van Stolberglaan 13, 1814 HB Alkmaar, The Netherlands.
authors.forename: Marion
authors.initials: M
authors.lastname: Smith
authors.affiliation: Department of Medical Microbiology, Medisch Centrum Alkmaar, Juliana Van Stolberglaan 13, 1814 HB Alkmaar, The Netherlands.|Regional Laboratory of Public Health, Haarlem 2035 RC, The Netherlands.
authors.forename: Bram
authors.initials: B
authors.lastname: Diederen
authors.affiliation: Department of Microbiology, University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Jelle
authors.initials: J
authors.lastname: Scharringa
authors.affiliation: Department of Medical Microbiology and Infection Prevention, Bronovo Hospital, The Hague, The Netherlands.
authors.forename: Maurine
authors.initials: M
authors.lastname: Leversteijn-van Hall
authors.affiliation: Department of Microbiology, University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Ad C
authors.initials: AC
authors.lastname: Fluit
authors.affiliation: Regional Laboratory of Public Health, Haarlem 2035 RC, The Netherlands.|Department of Medical Microbiology, Medisch Centrum Alkmaar, Juliana Van Stolberglaan 13, 1814 HB Alkmaar, The Netherlands.
authors.forename: James
authors.initials: J
authors.lastname: Cohen Stuart
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D057205:Automation, Laboratory / Q000379:methods; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics*; D004755:Enterobacteriaceae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D000077731:Meropenem; D008826:Microbial Sensitivity Tests; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013845:Thienamycins / Q000494:pharmacology; D013997:Time Factors; D001618:beta-Lactamases / Q000032:analysis* / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D013845:Thienamycins; D001618:beta-Lactamases; C063912:carbapenemase; D000077731:Meropenem