Subtyping and antifungal susceptibilities of Candida spp. in the intensive care unit of a Greek general hospital.

This study identified the Candida spp., susceptibility to antifungal agents and the prevailing Candida albicans subtypes responsible for infections or colonization of 42 patients in the ICU over a 6-month period. Most isolates were C. albicans (66.1%) and Candida tropicalis (28.3%) all of which were susceptible in vitro to antifungal agents. Subtypes of the C. albicans isolates were identified by pulsed field gel electrophoresis Sfi I chromosomal digests. Two major C. albicans subtypes were identified, whereas subtype heterogeneity was found among strains of Candida glabrata and C. tropicalis. Sfi I PFGE restriction patterns were able to discriminate between sub-populations of C. albicans isolates, clustering them into distinct, epidemiologically congruous groups.

Identification of medically significant fungal genera by polymerase chain reaction followed by restriction enzyme analysis.

Rapid non-culture-dependent assays for identification of fungi quicken diagnosis and prompt treatment of invasive fungal disease. Fungal DNA extracts from pure cultures of the most frequently isolated fungal pathogens belonging to the Genera Aspergillus, Candida and Cryptococcus along with less common pathogenic Genera were amplified with the general fungal primer pair internal transcribed spacer-1/4. Subsequently, the amplicon was digested with the restriction endonucleases MspI, HaeIII, HinfI and EcoRI in order to generate genus- or species-specific patterns for identification of the fungus. HinfI produced indistinguishable fingerprints for all Aspergillus species tested. MspI produced species-specific patterns for: Cryptococcus neoformans, Cryptococcus non-neoformans, Candida albicans and Candida tropicalis. EcoRI succeeded in differentiating penicillia from aspergilli and cryptococci from Candida spp. It is concluded that this procedure can differentiate genera and occasionally species of medically important fungi and that following the necessary validation experiments, it can be used directly on clinical samples to assist prompt diagnosis of systemic fungal infections.