Real-time TaqMan PCR for Rapid Detection and Typing of Genes Encoding CTX-M Extended-Spectrum Beta-Lactamases

J Med Microbiol. 2007 Jan;56(Pt 1):52-55. doi: 10.1099/jmm.0.46909-0.

Abstract

The prevalence of CTX-M-producing members of the Enterobacteriaceae is increasing worldwide. A novel, multiplex, real-time TaqMan PCR assay to detect and type bla(CTX-M) genes is described which is an improvement on previously described techniques with respect to reduced assay time, elimination of the need for protracted post-PCR processing and the convenience of a single reaction vessel. Based on beta-lactam antibiogram and MIC data, 478 of 1279 Enterobacteriaceae isolates from clinical blood and urine culture specimens were selected and tested for extended-spectrum beta-lactamase (ESBL) production using phenotypic methods. The new TaqMan assay detected and typed bla(CTX-M) genes in 21 of 28 ESBL-producing isolates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ampicillin / pharmacology
  • Anti-Bacterial Agents / pharmacology
  • Cephalexin / pharmacology
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / isolation & purification
  • Enterobacteriaceae / drug effects
  • Enterobacteriaceae / enzymology
  • Enterobacteriaceae / genetics*
  • Enterobacteriaceae Infections / blood
  • Enterobacteriaceae Infections / microbiology*
  • Enterobacteriaceae Infections / urine
  • Escherichia coli Proteins / genetics
  • Genotype
  • Humans
  • Microbial Sensitivity Tests
  • Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Sequence Analysis, DNA
  • beta-Lactam Resistance / genetics
  • beta-Lactamases / classification
  • beta-Lactamases / genetics*
  • beta-Lactamases / metabolism
  • beta-Lactams / pharmacology

Substances

  • Anti-Bacterial Agents
  • DNA, Bacterial
  • Escherichia coli Proteins
  • beta-Lactams
  • Ampicillin
  • CTX-M-9 protein, E coli
  • beta-lactamase CTX-M-15
  • beta-Lactamases
  • Cephalexin