title: Multi-centre evaluation of real-time multiplex PCR for detection of carbapenemase genes OXA-48, VIM, IMP, NDM and KPC.
abstract: BACKGROUND
  Resistance to carbapenem antibiotics is emerging worldwide among Enterobacteriaceae. To prevent hospital transmission due to unnoticed carriage of carbapenemase producing micro-organisms in newly admitted patients, or follow-up of patients in an outbreak setting, a molecular screening method was developed for detection of the most prevalent carbapenemase genes; blaOXA-48, blaVIM, blaIMP, blaNDM and blaKPC.
  
  
  METHODS
  A real-time multiplex PCR assay was evaluated using a collection of 86 Gram negative isolates, including 62 carbapenemase producers. Seven different laboratories carried out this method and used the assay for detection of the carbapenemase genes on a selection of 20 isolates.
  
  
  RESULTS
  Both sensitivity and specificity of the multiplex PCR assay was 100%, as established by results on the strain collection and the inter-laboratory comparisons.
  
  
  CONCLUSIONS
  In this study, we present a multiplex real-time PCR that is a robust, reliable and rapid method for the detection of the most prevalent carbapenemases blaOXA-48, blaVIM, blaIMP, blaNDM and blaKPC, and is suitable for screening of broth cultured rectal swabs and for identification of carbapenemase genes in cultures.
authors.affiliation: Maasstad Laboratory, Molecular Diagnostics Unit, Maasstad Hospital, Rotterdam, The Netherlands. Zeea@maasstadziekenhuis.nl.
authors.forename: Anneke
authors.initials: A
authors.lastname: van der Zee
authors.forename: Lieuwe
authors.initials: L
authors.lastname: Roorda
authors.forename: Gerda
authors.initials: G
authors.lastname: Bosman
authors.forename: Ad C
authors.initials: AC
authors.lastname: Fluit
authors.forename: Mirjam
authors.initials: M
authors.lastname: Hermans
authors.forename: Paul H M
authors.initials: PH
authors.lastname: Smits
authors.forename: Adri G M
authors.initials: AG
authors.lastname: van der Zanden
authors.forename: René
authors.initials: R
authors.lastname: Te Witt
authors.forename: Lesla E S
authors.initials: LE
authors.lastname: Bruijnesteijn van Coppenraet
authors.forename: James
authors.initials: J
authors.lastname: Cohen Stuart
authors.forename: Jacobus M
authors.initials: JM
authors.lastname: Ossewaarde
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D015780:Carbapenems; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004755:Enterobacteriaceae / Q000235:genetics*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D015780:Carbapenems; D001618:beta-Lactamases; C063912:carbapenemase