title: Molecular detection of Pseudomonas aeruginosa in recreational water.
abstract: The aim of this study was the development of a new molecular assay for Pseudomonas aeruginosa identification in recreational water. The method includes bacterial cell concentration through membrane filtration, a short (6 h) culture-enrichment step, DNA extraction and its amplification through a Real-Time PCR assay. The performance of the molecular approach was evaluated on 44 samples of swimming pool water and compared with the reference method UNI EN ISO 16266:2008. Positivity rates of 6% and 74% in pool and inlet water, respectively, with the standard culture method, and of 23% and 74% with the molecular method were found. Statistical analysis indicated "substantial agreement" (Cohen's Kappa index: 0.6831) between the two approaches. RAPD typing of P. aeruginosa isolates showed identical fingerprint profiles, indicating their epidemiological correlation. The developed protocol showed very high specificity and a detection limit of 10 genomic units. This technique has the potential to screen large numbers of environmental samples, and could be proposed as part of a self-monitoring plan for recreational facilities, improving surveillance and early warning systems.
authors.affiliation: Dipartimento di Scienze Biomolecolari, Università degli Studi di Urbino Carlo Bo, Urbino, Italy. giulia.amagliani@uniurb.it
authors.forename: Giulia
authors.initials: G
authors.lastname: Amagliani
authors.forename: Maria Lorella
authors.initials: ML
authors.lastname: Parlani
authors.forename: Giorgio
authors.initials: G
authors.lastname: Brandi
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Sebastianelli
authors.forename: Vilberto
authors.initials: V
authors.lastname: Stocchi
authors.forename: Giuditta Fiorella
authors.initials: GF
authors.lastname: Schiavano
mesh_terms: D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods*; D005374:Filtration; D008566:Membranes; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification*; D019105:Random Amplified Polymorphic DNA Technique; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013551:Swimming Pools; D014871:Water Microbiology
chemical_list: D004269:DNA, Bacterial