title: Development and validation of a molecular method for the diagnosis of medically important fungal infections.
abstract: The increasing incidence of severe fungal infections highlights the need for rapid and precise identification methods in clinical mycology. The aim of this study was to develop and validate a culture-indipendent molecular approach that could allow the detection of fungal pathogens in clinical samples, with particular attention to the identification of drug-resistant Candida and Aspergillus species. A real-time multiplex PCR assay was developed using TaqMan probes specific for highly discriminating ITS sequences. In its multiplex format the assay showed a high specificity, clearly discriminating among different species, as well as a high sensitivity (20 CFU/1 mL sample), making it a potentially useful starting point for the development of a more complete molecular diagnostic assay.
authors.affiliation: Laboratorio di Microbiologia e Virologia, Diagnostica e Ricerca San Raffaele, Milano. demarco.donata@hsr.it
authors.forename: Donata
authors.initials: D
authors.lastname: De Marco
authors.forename: Mario
authors.initials: M
authors.lastname: Perotti
authors.forename: Cristina M
authors.initials: CM
authors.lastname: Ossi
authors.forename: Roberto
authors.initials: R
authors.lastname: Burioni
authors.forename: Massimo
authors.initials: M
authors.lastname: Clementi
authors.forename: Nicasio
authors.initials: N
authors.lastname: Mancini
mesh_terms: D000935:Antifungal Agents; D001230:Aspergillus / Q000187:drug effects; D002175:Candida / Q000187:drug effects; D015342:DNA Probes; D004271:DNA, Fungal / Q000235:genetics; D021901:DNA, Intergenic; D004275:DNA, Ribosomal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D025141:Drug Resistance, Fungal; D005658:Fungi / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D009181:Mycoses / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019914:Taq Polymerase