title: Optimising the diagnostic strategy for onychomycosis from sample collection to FUNGAL identification evaluation of a diagnostic kit for real-time PCR.
abstract: Onychomycosis is a common nail disorder mainly due to dermatophytes for which the conventional diagnosis requires direct microscopic observation and culture of a biological sample. Nevertheless, antifungal treatments are commonly prescribed without a mycological examination having been performed, partly because of the slow growth of dermatophytes. Therefore, molecular biology has been applied to this pathology, to support a quick and accurate distinction between onychomycosis and other nail damage. Commercial kits are now available from several companies for improving traditional microbiological diagnosis. In this paper, we present the first evaluation of the real-time PCR kit marketed by Bio Evolution for the diagnosis of dermatophytosis. Secondly, we compare the efficacy of the kit on optimal and non-optimal samples. This study was conducted on 180 nails samples, processed by conventional methods and retrospectively analysed using this kit. According to our results, this molecular kit has shown high specificity and sensitivity in detecting dermatophytes, regardless of sample quality. On the other hand, and as expected, optimal samples allowed the identification of a higher number of dermatophytes by conventional mycological diagnosis, compared to non-optimal samples. Finally, we have suggested several strategies for the practical use of such a kit in a medical laboratory for quick pathogen detection.
authors.affiliation: Structure de Parasitologie-Mycologie, Département de Microbiologie, Centre Hospitalo-Universitaire de Nancy (CHU-Nancy), Hôpitaux de Brabois, Vandœuvre-les-Nancy, France.
authors.forename: Dimitri
authors.initials: D
authors.lastname: Petinataud
authors.affiliation: Plateforme de génomique microbienne, Département de Microbiologie, CHU-Nancy, Hôpitaux de Brabois, Vandœuvre-les-Nancy, France.
authors.forename: Sibel
authors.initials: S
authors.lastname: Berger
authors.affiliation: Service de Biostatistiques, CHU Félix Guyon, la Réunion, Centre de Méthodologie et de Gestion des données: Unité de Soutien Méthodologique, CHU La Réunion-Site du CHFG Route de Bellepierre, Saint Denis Cedex, France.
authors.forename: Cyril
authors.initials: C
authors.lastname: Ferdynus
authors.affiliation: Structure de Parasitologie-Mycologie, Département de Microbiologie, Centre Hospitalo-Universitaire de Nancy (CHU-Nancy), Hôpitaux de Brabois, Vandœuvre-les-Nancy, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Debourgogne
authors.affiliation: Structure de Parasitologie-Mycologie, Département de Microbiologie, Centre Hospitalo-Universitaire de Nancy (CHU-Nancy), Hôpitaux de Brabois, Vandœuvre-les-Nancy, France.
authors.forename: Nelly
authors.initials: N
authors.lastname: Contet-Audonneau
authors.affiliation: Structure de Parasitologie-Mycologie, Département de Microbiologie, Centre Hospitalo-Universitaire de Nancy (CHU-Nancy), Hôpitaux de Brabois, Vandœuvre-les-Nancy, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Machouart
mesh_terms: D000368:Aged; D003883:Arthrodermataceae / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D004271:DNA, Fungal / Q000737:chemistry / Q000302:isolation & purification; D005260:Female; D005658:Fungi / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D014009:Onychomycosis / Q000175:diagnosis* / Q000382:microbiology; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000592:standards; D013997:Time Factors
chemical_list: D004271:DNA, Fungal; D011933:Reagent Kits, Diagnostic
keywords: Onychomycosis; PCR; culture; diagnosis; direct examination; kit