Real-time TaqMan PCR for rapid detection and typing of genes encoding CTX-M extended-spectrum β-lactamases

• Published in: Journal of medical microbiology Vol. 56; no. 1; pp. 52 - 55
• Main Authors: BIRKETT, Christopher I, LUDLAM, Hugo A, WOODFORD, Neil, BROWN, Derek F. J, BROWN, Nicholas M, ROBERTS, Mark T. M, MILNER, Nicola, CURRAN, Martin D
• Format: Journal Article
• Language: English
• Published: Reading Society for General Microbiology 2007
• Subjects: Ampicillin - pharmacology; Anti-Bacterial Agents - pharmacology; beta-Lactam Resistance - genetics; beta-Lactamases - classification; beta-Lactamases - genetics; beta-Lactamases - metabolism; beta-Lactams - pharmacology; Biological and medical sciences; Cephalexin - pharmacology; DNA, Bacterial - chemistry; DNA, Bacterial - isolation & purification; Enterobacteriaceae; Enterobacteriaceae - drug effects; Enterobacteriaceae - enzymology; Enterobacteriaceae - genetics; Enterobacteriaceae Infections - blood; Enterobacteriaceae Infections - microbiology; Enterobacteriaceae Infections - urine; Escherichia coli Proteins - genetics; Fundamental and applied biological sciences. Psychology; Genotype; Humans; Infectious diseases; Medical sciences; Microbial Sensitivity Tests; Microbiology; Polymerase Chain Reaction - methods; Reproducibility of Results; Sequence Analysis, DNA; Polymerase chain reaction; Typing; Gene; Microbiology; Extended-spectrum β-lactamase; Detection; Real time
• ISSN: 0022-2615; 1473-5644
• DOI: 10.1099/jmm.0.46909-0

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.