title: Contribution of molecular tools for the diagnosis and epidemiology of fungal chronic rhinosinusitis.
abstract: Chronic rhinosinusitis (CRS) rank second at chronic inflammatory diseases in industrialized countries and are an important public health concern. Diagnosis relies on a set of arguments including clinical signs, imaging, histopathologic and mycological analyses of sinus specimens, collected during nasal endoscopy. The sensitivity of fungal cultures is reported to be poor, even when direct examination is positive, thus the epidemiology of fungal chronic sinusitis is ill-known. This study evaluated the sensitivity of molecular diagnosis in 70 consecutive samples (61 patients with CRS) analysed at the University Hospital of Rennes during a 3-year period. DNA detection was performed using a conventional PCR method targeting the ITS1/ITS2 sequence and the resulting amplification products were sequenced. Fungal CRS was proven in 42 patients (69%), of which only 20 (48%) had a positive culture. 37/42 (88%) patients were diagnosed with a fungus ball, 3 with allergic fungal CRS and 2 with undetermined fungal CRS. PCR was positive in all 42 cases and direct sequencing allowed to identify fungi in all cases but one, and detected multiple infection in 3. Aspergillus fumigatus was present in 69% of patients; Cladosporium cladosporoides in 9.5%, Scedosporium sp., A. nidulans and A. flavus in 7% each. In 2/19 patients with negative direct examination, sequencing analysis revealed the presence of Capnobotryella sp. and C. cladosporoides, in clinical settings compatible with fungal sinusitis. In conclusion, ITS1/ITS2 PCR had a twice better sensitivity than culture, and combined sequencing provides accurate epidemiological data on fungal CRS.
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, France.
authors.forename: Pauline
authors.initials: P
authors.lastname: Comacle
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, France Inserm U1085-IRSET, Université Rennes 1, Rennes, France.
authors.forename: Sorya
authors.initials: S
authors.lastname: Belaz
authors.affiliation: Service d'Otorhino-laryngologie, Centre Hospitalier Universitaire de Rennes, Rennes, France.
authors.forename: Franck
authors.initials: F
authors.lastname: Jegoux
authors.affiliation: Clinique Mutualiste La Sagesse, Rennes, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Ruaux
authors.affiliation: Service d'Anatomo-Pathologie, Centre Hospitalier Universitaire de Rennes, Rennes, France.
authors.forename: François
authors.initials: F
authors.lastname: Le Gall
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, France Inserm U1085-IRSET, Université Rennes 1, Rennes, France.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Gangneux
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, France Inserm U1085-IRSET, Université Rennes 1, Rennes, France Florence.robert-gangneux@univ-rennes1.fr.
authors.forename: Florence
authors.initials: F
authors.lastname: Robert-Gangneux
mesh_terms: D002908:Chronic Disease; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D005260:Female; D005658:Fungi / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006785:Hospitals, University; D006801:Humans; D008297:Male; D008828:Microbiological Techniques / Q000379:methods; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D017720:Molecular Epidemiology / Q000379:methods; D009181:Mycoses / Q000175:diagnosis* / Q000453:epidemiology; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies; D012220:Rhinitis / Q000175:diagnosis* / Q000453:epidemiology; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012852:Sinusitis / Q000175:diagnosis* / Q000453:epidemiology
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer
keywords: ITS1/ITS2; PCR; chronic rhinosinusitis; fungi; sequence analysis