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

BMC Infectious Diseases

Jacobus M Ossewaarde⁵  James Cohen Stuart⁴  Lesla ES Bruijnesteijn van Coppenraet⁷  René te Witt⁵  Adri GM van der Zanden³  Paul HM Smits¹  Mirjam Hermans²  Ad C Fluit⁶  Gerda Bosman⁸  Lieuwe Roorda⁸  Anneke van der Zee⁸ 

[1] Molecular Biology Laboratory, Slotervaart Hospital, Amsterdam, The Netherlands;Molecular Diagnostics, Jeroen Bosch Hospital, ‘s-Hertogenbosch, The Netherlands;Laboratory for Microbiology and Public Health, Enschede, The Netherlands;Medical Centre Alkmaar, Alkmaar, The Netherlands;Department of Medical Microbiology and Infectious Diseases, Erasmus MC, Rotterdam, The Netherlands;Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands;Isala clinics, Laboratory for Medical Microbiology and Infectious Diseases, Zwolle, The Netherlands;Maasstad Laboratory, Molecular Diagnostics Unit, Maasstad Hospital, Rotterdam, The Netherlands

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

Others  :  1134998
DOI  :  10.1186/1471-2334-14-27

 received in 2013-09-03, accepted in 2013-12-24,  发布年份 2014

【 摘 要 】

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; bla_OXA-48, bla_VIM, bla_IMP, bla_NDM and bla_KPC.

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 bla_OXA-48, bla_VIM, bla_IMP, bla_NDM and bla_KPC, and is suitable for screening of broth cultured rectal swabs and for identification of carbapenemase genes in cultures.

【 授权许可】

   
2014 van der Zee et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

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