title: Using newly developed multiplex polymerase chain reaction and melting curve analysis for detection and discrimination of β-lactamases in Escherichia coli isolates from intensive care patients.
abstract: A total of 78 bacterial strains with known β-lactamases were used to optimize a rapid detection system consisting of multiplex polymerase chain reaction and melting curve analysis to amplify and identify blaTEM, blaSHV, and blaCTX-M genes in a single reaction. Additionally, to evaluate the applicability of this method, 32 clinical isolates of Escherichia coli displaying an extended-spectrum β-lactamase phenotype from patients hospitalized at intensive care units were tested. Results were analyzed by the Rotor-Gene operating software and Rotor-Gene ScreenClust HRM Software. The individual melting curves differed by a temperature shift or curve shape, according to the presence of β-lactamase genes. With the use of this method and direct sequencing, blaCTX-M-15-like was identified as the most prevalent β-lactamase gene. In conclusion, this novel detection system seems to be a suitable tool for rapid detection of present β-lactamase genes and their characterization.
authors.affiliation: Department of Microbiology, Faculty of Medicine and Dentistry, Palacký University in Olomouc, Czech Republic. magdalena.chroma@centrum.cz
authors.forename: Magdaléna
authors.initials: M
authors.lastname: Chromá
authors.forename: Kristýna
authors.initials: K
authors.lastname: Hricová
authors.forename: Milan
authors.initials: M
authors.lastname: Kolář
authors.forename: Pavel
authors.initials: P
authors.lastname: Sauer
authors.forename: Dagmar
authors.initials: D
authors.lastname: Koukalová
mesh_terms: D003428:Cross Infection / Q000382:microbiology; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000382:microbiology; D006801:Humans; D007362:Intensive Care Units; D060885:Multiplex Polymerase Chain Reaction; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D001618:beta-Lactamases