부산대학교 도서관

Objective: To compare the conventional identification with the molecular methods, based on fungal DNA markers. Methods: 95 strains were isolated at SVB between Jan-Apr 2009. Their isolation was made on BHI sheep blood/agar, Sabouraud agar, ChromID Candida agar. Yeasts were identified according to Germ tube test, India ink examination, rapid urease test, detection of soluble Cryptococcus neoformans antigens, API ID 32 C and VITEK 2C. At the CTMA-UCL 30 DNA extractions were made with NucliSENS® lysis magnetic extraction reagents and NucliSENS® miniMAG. A real time PCR amplification was performed using the ITS3 and ITS4 primers. Data were recorded on a Roche LightCycler® 480 System. Aspergillus fumigatus strain DSM 63359 was used as a positive control. The amplicons were sequenced on an automated 3130 Genetic Analyser. Results: The strains identified by conventional methods in SVB were: C. albicans, C. glabrata, C. krusei, Sacharomices cerevisiae, C. kefyr, C. tropicalis, C. guillermondii, C. holmii, Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus spp, Cryptococcus neoformans. Correlation between the melting genotype and the phenotypic identification was perfect for C. albicans, C. glabrata, C. krusei, C. guillermondii and C. neoformans. Conclusions: SVB place a high priority on maximizing their capabilities for the early diagnosis of opportunistic fungal infections. [ABSTRACT FROM AUTHOR]