A rapid PCR-RFLP assay for identification of 11 medically important Candida species, Trichosporon asahii, Cryptococcus neoformans, Saccharomyces cerevisiae, Hansonula anomala and Geotrichum candidum

Abstract number: 1134_03_251

Mirhendi H., Makimura K.

Opportunistic yeast infections have various clinical features and are caused by several Candida species and by several other yeasts belong to other genera. The emergence of these mycoses is increasing, for example deep-seated candidiasis has increased dramatically in recent years. Rapid identification of yeast isolates to the species level in the clinical laboratory is necessary to more rapid and effective antifungal therapy and to facilitate hospital infection control measures. Conventional phenotype-based methods for identifying yeasts species are often difficult and time consuming. Molecular biological techniques provide useful alternative methods. In this study, ITS1-5.8S-ITS2 region of the fungal rRNA genes were amplified in 31 yeast standard strain and 137 clinical Candida isolates with the universal primers ITS1 and ITS4. Digestion the PCR product by the restriction enzymes HpaII and AcsI allowed us to clearly identify C. albicans, C. glabrata, C. parapsilosis, AC. tropicalis, AC. krusei, C. dubliniensis, C. guilliermondii, C. lusitania, C. rogosa, C. famata, C. kefyr, Trichosporonasahii, Cryptococcusneoformans, Saccharomycescerevisiae, Hansonulaanomala and Geotrichumcandidum. All clinical isolates were checked by cultivation on Chrome-agar Candida. This panel of PCR-restriction enzyme could be rapid, simple and useful in diagnostic and epidemiological studies of Candida and candidiasis and the infections caused by other yeasts, without any requirement of expensive equipments.