Identyfikacja grzybów z rodzaju Candida na podstawie analizy polimorfizmu fragmentów restrykcyjnych rybosomalnego DNA

Introduction: The identification of microorganism represent a key part of microbiological investigation, which is especially important in case of Candida species intristincly resistant to antimycotics. Phenotypic methods of identification are frequently not enough specific and simple molecular tests of high specificity are expected. Purpose: Restriction analysis of selected rDNA sequences of clinically relevant Candida species and evaluation their potential to be used in the RFLP method for the construction of diagnostic tests Material and methods: The fragments of rDNA flanked by primers ITS1 and ITS4, and NS3 and ITS4 of forteen Candida species were analysed. The sequences were obtained from PubMed-NCBI (www.ncbi.nlm. nih.gov) and analised for restriction with HaeIII and ScrFI enzymes, with the help of Webcutter 2.0 [http:// rna.lundberg.gu.se/cutter2/]. Results: Analysis of the length of ITS1-ITS4 flanked rDNA fragment, together with restriction analysis with HaeIII differentiate eleven and together with ScrFI twelve species. Similarly, restrictions patterns of DNA fragments flanked by NS3-ITS4 could distinguish 12/14 species. RFLP with HaeIII and ScrFI enzymes cannot differentiate between C. albicans and C. dubliniensis. Conclusion: 1. The use of ScrFI and HaeIII enzymes for analysis of rDNA fragments flanked with primers pars ITS1 and ITS4, and NS3 and ITS4, give the possibility to differentiate twelve Candida species. 2. The distinguishing between C. albicans and C. dubliniensis could be performed with the use of additional restricted nucleases. 3. Presented data should be confirmed experimentally.