title: Real-time TaqMan PCR for rapid detection and typing of genes encoding CTX-M extended-spectrum beta-lactamases.
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.
authors.affiliation: Health Protection Agency, Clinical Microbiology and Public Health Laboratory, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Christopher I
authors.initials: CI
authors.lastname: Birkett
authors.affiliation: Health Protection Agency, Clinical Microbiology and Public Health Laboratory, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Hugo A
authors.initials: HA
authors.lastname: Ludlam
authors.affiliation: Antibiotic Resistance Monitoring and Reference Laboratory, Centre for Infections, Health Protection Agency, London, UK.
authors.forename: Neil
authors.initials: N
authors.lastname: Woodford
authors.affiliation: Health Protection Agency, Clinical Microbiology and Public Health Laboratory, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Derek F J
authors.initials: DFJ
authors.lastname: Brown
authors.affiliation: Health Protection Agency, Clinical Microbiology and Public Health Laboratory, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Nicholas M
authors.initials: NM
authors.lastname: Brown
authors.affiliation: Health Protection Agency, Clinical Microbiology and Public Health Laboratory, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Mark T M
authors.initials: MTM
authors.lastname: Roberts
authors.affiliation: Health Protection Agency, Clinical Microbiology and Public Health Laboratory, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Nicola
authors.initials: N
authors.lastname: Milner
authors.affiliation: Health Protection Agency, Clinical Microbiology and Public Health Laboratory, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Martin D
authors.initials: MD
authors.lastname: Curran
mesh_terms: D000667:Ampicillin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002506:Cephalexin / Q000494:pharmacology; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D004755:Enterobacteriaceae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D004756:Enterobacteriaceae Infections / Q000097:blood / Q000382:microbiology* / Q000652:urine; D029968:Escherichia coli Proteins / Q000235:genetics; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D018440:beta-Lactam Resistance / Q000235:genetics; D001618:beta-Lactamases / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D047090:beta-Lactams / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D047090:beta-Lactams; D000667:Ampicillin; C412613:CTX-M-9 protein, E coli; C473109:beta-lactamase CTX-M-15; D001618:beta-Lactamases; D002506:Cephalexin