title: Direct identification of major Gram-negative pathogens in respiratory specimens by respiFISH® HAP Gram (-) Panel, a beacon-based FISH methodology.
abstract: Rapid detection of microorganisms in respiratory specimens is of paramount importance to drive the proper antibiotic regimen to prevent complications and transmission of infections. In the present study, the respiFISH® HAP Gram (-) Panel (miacom diagnostics GmbH, Duesseldorf, Germany) for the etiological diagnosis of hospital-acquired pneumonia was compared with the traditional culture method for the detection of major Gram-negative pathogens in respiratory specimens. respiFISH® combined the classical fluorescence in situ hybridization (FISH) technology with fluorescence-labeled DNA molecular beacons as probes. From September 2011 to January 2012, 165 samples were analyzed: the sensitivity and specificity were 94.39 and 87.93%, respectively. Only six pathogens (3.6%) were not identified with respiFISH®, while seven specimens (3%) provided false-positive results. This beacon-based identification shortens the time to result by at least one work day, providing species-level identification within half an hour. Considering the high sensitivity and specificity and the significant time saving, the introduction of bbFISH® assays could effectively complement traditional systems in microbiology laboratories.
authors.affiliation: Department of Pathology and Diagnostics, University of Verona, Strada Le Grazie 8, 37134, Verona, Italy. raffaella.koncan@univr.it.
authors.forename: R
authors.initials: R
authors.lastname: Koncan
authors.affiliation: O.U. of Microbiology and Virology, Department of Pathology and Diagnostics, Azienda Ospedaliera Universitaria Integrata di Verona, P.le A. Scuro 10, 37134, Verona, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Parisato
authors.affiliation: Clinical Microbiology Laboratories, Foundation Polyclinic Tor Vergata, V.le Oxford 81, 00133, Roma, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Sakarikou
authors.affiliation: Department of Anesthesiological Sciences and Specialistic Surgeries, University of Verona, P.le A. Scuro 10, 37134, Verona, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Stringari
authors.affiliation: Clinical Microbiology Laboratories, Foundation Polyclinic Tor Vergata, V.le Oxford 81, 00133, Roma, Italy.|Department of Experimental Medicine and Surgery, "Tor Vergata" University of Rome, via Montpellier 1, 00133, Roma, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Fontana
authors.affiliation: Department of Pathology and Diagnostics, University of Verona, Strada Le Grazie 8, 37134, Verona, Italy.
authors.forename: V
authors.initials: V
authors.lastname: Favuzzi
authors.affiliation: Department of Pathology and Diagnostics, University of Verona, Strada Le Grazie 8, 37134, Verona, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Ligozzi
authors.affiliation: O.U. of Microbiology and Virology, Department of Pathology and Diagnostics, Azienda Ospedaliera Universitaria Integrata di Verona, P.le A. Scuro 10, 37134, Verona, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Lo Cascio
mesh_terms: D046508:Culture Techniques; D016905:Gram-Negative Bacterial Infections / Q000145:classification* / Q000175:diagnosis*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D011014:Pneumonia / Q000175:diagnosis* / Q000382:microbiology*; D012680:Sensitivity and Specificity