, Volume 48, Issue 2, pp 233-238

PrimersITS1, ITS2 andITS4 detect the intraspecies variability in the internal transcribed spacers and 5.8S rRNA gene region in clinical isolates of fungi

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Abstract

Restriction fragment length polymorphism analysis of the 5.8S rRNA gene and the internal transcribed spacers (ITS1 andITS2) was used for examination of 66 isolates belonging to 19 species. Intraspecies variability was found in the examined region of 11 species (Candida albicans, C. catenulata, C. colliculosa, C. glabrata, C. kefyr, C. melinii, C. parapsilosis, C. guillermondii, C. solanii, C. tropicalis, Saccharomyces cerevisiae). Region of ITS-5.8S rDNA was amplified using the primersITS1 andITS4. The amplicons were digested byHaeIII,HinfI andCfoI. The recognized intraspecies variability was confirmed in the second step, in which the shorter fragments of this region were amplified using primersITS1 andITS2 and analyzed by capillary electrophoresis. Considerable intraspecific variability renders this method unsuitable for species identification, whereas it can be useful for epidemiological tracing of isolates.

This work was supported by grant no.117/1999C of theGrant Agency of Charles University and by the research project no. CEZ 111 400 002 of theMinistry of Education, Youth and Sports of the Czech Republic.