title: Application of Molecular Diagnostics in Primary Detection of ESBL Directly from Clinical Specimens.
abstract: The infections caused by extended-spectrum β-lactamase (ESBL)-producing organisms are associated with increased mortality. The real-time polymerase chain reaction (PCR) method, which enables detection of ESBLs directly from patients' clinical material, was developed. This study focused on blaCTX-M and blaSHV determination in endotracheal aspirates. Each sample was identified with standard microbiological procedures and simultaneously analyzed for the presence of nucleic acids, which encode CTX-M and SHV ESBL enzymes using real-time PCR. A total of 341 samples were investigated. In the set, 27 ESBL-positive samples were identified by phenotypic methods, while 60 positive samples were identified by the PCR method. Of the 60 PCR-positive samples, 58 were positive for the blaCTX-M. In two samples, the ESBL blaSHV-ESBL gene was detected. One phenotypically positive sample was PCR negative. The real-time PCR assay does not require a cultivation step and therefore enables detection of ESBL in 6 hours. The rapid method is necessary for early and adequate antimicrobial treatment.
authors.affiliation: 1Department of Microbiology, Faculty of Medicine, St. Anne's University Hospital, Masaryk University, Brno, Czech Republic.|2GeneProof a.s., Brno, Czech Republic.
authors.forename: Martina
authors.initials: M
authors.lastname: Sittová
authors.affiliation: 3Department of Microbiology, Faculty of Medicine and Dentistry, Palacký University Olomouc, Olomouc, Czech Republic.
authors.forename: Magdaléna
authors.initials: M
authors.lastname: Röderová
authors.affiliation: 2GeneProof a.s., Brno, Czech Republic.
authors.forename: Miloš
authors.initials: M
authors.lastname: Dendis
authors.affiliation: 3Department of Microbiology, Faculty of Medicine and Dentistry, Palacký University Olomouc, Olomouc, Czech Republic.
authors.forename: Kristýna
authors.initials: K
authors.lastname: Hricová
authors.affiliation: 3Department of Microbiology, Faculty of Medicine and Dentistry, Palacký University Olomouc, Olomouc, Czech Republic.
authors.forename: Vendula
authors.initials: V
authors.lastname: Pudová
authors.affiliation: 2GeneProof a.s., Brno, Czech Republic.
authors.forename: Radek
authors.initials: R
authors.lastname: Horváth
authors.affiliation: 1Department of Microbiology, Faculty of Medicine, St. Anne's University Hospital, Masaryk University, Brno, Czech Republic.
authors.forename: Filip
authors.initials: F
authors.lastname: Růžička
authors.affiliation: 2GeneProof a.s., Brno, Czech Republic.
authors.forename: Šárka
authors.initials: Š
authors.lastname: Dosoudilová
authors.affiliation: 3Department of Microbiology, Faculty of Medicine and Dentistry, Palacký University Olomouc, Olomouc, Czech Republic.
authors.forename: Milan
authors.initials: M
authors.lastname: Kolář
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004755:Enterobacteriaceae / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D004756:Enterobacteriaceae Infections / Q000175:diagnosis* / Q000188:drug therapy / Q000382:microbiology; D015870:Gene Expression; D006801:Humans; D008826:Microbial Sensitivity Tests; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000188:drug therapy / Q000382:microbiology; D012189:Retrospective Studies; D018440:beta-Lactam Resistance / Q000235:genetics*; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001618:beta-Lactamases