Multiplex PCR for rapid detection of genes encoding class A carbapenemases

Ann Lab Med. 2012 Sep;32(5):359-61. doi: 10.3343/alm.2012.32.5.359. Epub 2012 Aug 13.

Abstract

In recent years, there have been increasing reports of KPC-producing Klebsiella pneumoniae in Korea. The modified Hodge test can be used as a phenotypic screening test for class A carbapenamase (CAC)-producing clinical isolates; however, it does not distinguish between carbapenemase types. The confirmation of type of CAC is important to ensure optimal therapy and to prevent transmission. This study applied a novel multiplex PCR assay to detect and differentiate CAC genes in a single reaction. Four primer pairs were designed to amplify fragments encoding 4 CAC families (SME, IMI/NMC-A, KPC, and GES). The multiplex PCR detected all genes tested for 4 CAC families that could be differentiated by fragment size according to gene type. This multiplex PCR offers a simple and useful approach for detecting and distinguishing CAC genes in carbapenem-resistant strains that are metallo-β-lactamase nonproducers.

Keywords: Carbapenemase; GES; KPC; Multiplex PCR.

MeSH terms

  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • DNA Primers / metabolism
  • Databases, Genetic
  • Humans
  • Klebsiella Infections / microbiology
  • Klebsiella pneumoniae / genetics
  • Klebsiella pneumoniae / isolation & purification
  • Klebsiella pneumoniae / metabolism
  • Multiplex Polymerase Chain Reaction*
  • beta-Lactamases / genetics*
  • beta-Lactamases / metabolism

Substances

  • Bacterial Proteins
  • DNA Primers
  • beta-Lactamases
  • carbapenemase