title: Development of a multiplex real-time PCR for the rapid detection of the predominant beta-lactamase genes CTX-M, SHV, TEM and CIT-type AmpCs in Enterobacteriaceae.
abstract: Beta-lactamase resistant bacteria and especially ESBL producing Enterobacteriaceae are an increasing problem worldwide. For this reason a major interest in efficient and reliable methods for rapid screening of high sample numbers is recognizable. Therefore, a multiplex real-time PCR was developed to detect the predominant class A beta-lactamase genes blaCTX-M, blaSHV, blaTEM and CIT-type AmpCs in a one-step reaction. A set of 114 Enterobacteriaceae containing previously identified resistance gene subtypes and in addition 20 undefined animal and environmental isolates were used for the validation of this assay. To confirm the accessibility in variable settings, the real-time runs were performed analogous in two different laboratories using different real-time cyclers. The obtained results showed complete accordance between the real-time data and the predetermined genotypes. Even if sequence analyses are further necessary for a comprehensive characterization, this method was proofed to be reliable for rapid screening of high sample numbers and therefore could be an important tool for e. g. epidemiological purposes or support infection control measures.
authors.affiliation: Institute of Animal Hygiene and Environmental Health, Free University Berlin, Berlin, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Roschanski
authors.affiliation: Federal Institute for Risk Assessment, Department of Biological Safety, Berlin, Germany.
authors.forename: Jennie
authors.initials: J
authors.lastname: Fischer
authors.affiliation: Federal Institute for Risk Assessment, Department of Biological Safety, Berlin, Germany.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Guerra
authors.affiliation: Institute of Animal Hygiene and Environmental Health, Free University Berlin, Berlin, Germany.
authors.forename: Uwe
authors.initials: U
authors.lastname: Roesler
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D004755:Enterobacteriaceae / Q000187:drug effects* / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics; D008826:Microbial Sensitivity Tests; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C048795:AmpC beta-lactamases; D001618:beta-Lactamases; C000602253:beta-lactamase CTX-M, E coli