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