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Open Access Highly Accessed Research article

Multi-centre evaluation of real-time multiplex PCR for detection of carbapenemase genes OXA-48, VIM, IMP, NDM and KPC

Anneke van der Zee1*, Lieuwe Roorda1, Gerda Bosman1, Ad C Fluit2, Mirjam Hermans3, Paul HM Smits4, Adri GM van der Zanden5, René te Witt6, Lesla ES Bruijnesteijn van Coppenraet7, James Cohen Stuart8 and Jacobus M Ossewaarde16

Author Affiliations

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

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BMC Infectious Diseases 2014, 14:27  doi:10.1186/1471-2334-14-27

Published: 14 January 2014

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.

Keywords:
Real-time multiplex PCR; Carbapenemases; OXA-48; VIM; IMP; NDM; KPC