1. Real-time TaqMan PCR for rapid detection and typing of genes encoding CTX-M extended-spectrum β-lactamases
- Author
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Christopher I. Birkett, Nicholas M. Brown, Mark T. M. Roberts, Nicola J. Milner, Hugo A. Ludlam, Martin D. Curran, Neil Woodford, and Derek F. J. Brown
- Subjects
DNA, Bacterial ,Microbiology (medical) ,Genotype ,Sequence analysis ,Microbial Sensitivity Tests ,Biology ,beta-Lactams ,Polymerase Chain Reaction ,Microbiology ,beta-Lactam Resistance ,beta-Lactamases ,law.invention ,Enterobacteriaceae ,law ,TaqMan ,Humans ,Multiplex ,Typing ,Gene ,Polymerase chain reaction ,Cephalexin ,Escherichia coli Proteins ,Enterobacteriaceae Infections ,Reproducibility of Results ,Sequence Analysis, DNA ,General Medicine ,biochemical phenomena, metabolism, and nutrition ,bacterial infections and mycoses ,biology.organism_classification ,Anti-Bacterial Agents ,Ampicillin - 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 β-lactam antibiogram and MIC data, 478 of 1279 Enterobacteriaceae isolates from clinical blood and urine culture specimens were selected and tested for extended-spectrum β-lactamase (ESBL) production using phenotypic methods. The new TaqMan assay detected and typed bla CTX-M genes in 21 of 28 ESBL-producing isolates.
- Published
- 2007
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