Multi-centre evaluation of real-time multiplex PCR for detection of carbapenemase genes OXA-48, VIM, IMP, NDM and KPC
1 Maasstad Laboratory, Molecular Diagnostics Unit, Maasstad Hospital, Rotterdam, The Netherlands
2 Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands
3 Molecular Diagnostics, Jeroen Bosch Hospital, ‘s-Hertogenbosch, The Netherlands
4 Molecular Biology Laboratory, Slotervaart Hospital, Amsterdam, The Netherlands
5 Laboratory for Microbiology and Public Health, Enschede, The Netherlands
6 Department of Medical Microbiology and Infectious Diseases, Erasmus MC, Rotterdam, The Netherlands
7 Isala clinics, Laboratory for Medical Microbiology and Infectious Diseases, Zwolle, The Netherlands
8 Medical Centre Alkmaar, Alkmaar, The Netherlands
BMC Infectious Diseases 2014, 14:27 doi:10.1186/1471-2334-14-27Published: 14 January 2014
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.
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.
Both sensitivity and specificity of the multiplex PCR assay was 100%, as established by results on the strain collection and the inter-laboratory comparisons.
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.