title: Validation of a new seminested PCR-based detection method for Legionella pneumophila.
abstract: We report a new seminested PCR method for detection of Legionella pneumophila based on the simultaneous amplification of a 387 bp fragment of the dotA gene and a 827 bp recombinant fragment that serves as an internal positive control. This new detection system was validated and its specificity and detection limit were determined. Parallel analysis of 90 environmental samples to compare this method, a real-time PCR method and the standard culture isolation method, demonstrated that this seminested method is useful for the study of L. pneumophila.
authors.affiliation: Applus+ LABAQUA, Pol Ind. Atalayas, C) Del Dracma 16-18, 03114 Alicante, Spain.
authors.forename: M A
authors.initials: MA
authors.lastname: Yáñez
authors.forename: V M
authors.initials: VM
authors.lastname: Barberá
authors.forename: V
authors.initials: V
authors.lastname: Catalán
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001431:Bacteriological Techniques; D002352:Carrier Proteins / Q000235:genetics; D016952:Legionella pneumophila / Q000235:genetics / Q000302:isolation & purification*; D008565:Membrane Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D012680:Sensitivity and Specificity; D014871:Water Microbiology
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; C440002:DotA protein, Legionella pneumophila; D008565:Membrane Proteins