A multiplex real-time PCR for the direct, fast, economic and simultaneous detection of the carbapenemase genes blaKPC, blaNDM, blaVIM and blaOXA-48

J Microbiol Methods. 2017 Nov:142:20-26. doi: 10.1016/j.mimet.2017.08.017. Epub 2017 Aug 24.

Abstract

A novel multiplex real-time PCR was designed to detect the clinically most important carbapenemase genes, blaKPC, blaVIM, blaNDM and blaOXA-48. The multiplex assay was verified testing genomic DNA of 24 carbapenemase-producing strains. It was validated using a blinded panel of 82 carbapenemase-producing and 50 non-producing isolates by direct colony PCR.

Keywords: CPO; Carbapenemases; Colony PCR; Hospital; Microarray; Multiplex; Real-time PCR.

MeSH terms

  • Bacterial Proteins / genetics*
  • Base Sequence
  • Carbapenem-Resistant Enterobacteriaceae / drug effects*
  • Carbapenem-Resistant Enterobacteriaceae / genetics*
  • Carbapenem-Resistant Enterobacteriaceae / isolation & purification
  • Carbapenems / pharmacology
  • Genotype
  • Microbial Sensitivity Tests / methods
  • Real-Time Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • beta-Lactamases / genetics*

Substances

  • Bacterial Proteins
  • Carbapenems
  • beta-Lactamases
  • carbapenemase