Development and validation of a molecular method for the diagnosis of medically important fungal infections.
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.
De Marco
Donata
D
Laboratorio di Microbiologia e Virologia, Diagnostica e Ricerca San Raffaele, Milano. demarco.donata@hsr.it
Perotti
Mario
M
Ossi
Cristina M
CM
Burioni
Roberto
R
Clementi
Massimo
M
Mancini
Nicasio
N
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