The aim of this study was to evaluate the use of one of the molecular typing methods such as PCR (polymerase chain reaction) following by RFLP (restriction fragment length polymorphism) analysis in the identification of
Candida
species and then to differentiate the identified azole susceptible and resistant
Candida albicans strains by using AP-PCR (arbitrarily primed-polymerase chain reaction). The identification of
Candida species by PCR and RFLP analysis was based on the size and primary structural variation of rDNA intergenic spacer regions (ITS). Forty-four clinical
Candida isolates comprising 5 species were included to the study. The amplification products were digested individually with 3 different restriction enzymes:
HaeIII,
DdeI, and
BfaI. All the isolates tested yielded the expected band patterns by PCR and RFLP analysis. The results obtained from this study demonstrate that
Candida species can be differentiated as
C. albicans and non-
C. albicans strains only by using
HaeIII restriction enzyme and
BfaI maintains the differentiation of these non-
C. albicans species. After identification
Candida species with RFLP analysis,
C. albicans strains were included to the AP-PCR test. By using AP-PCR, fluconazole susceptible and resistant strains were differentiated. Nine fluconazole susceptible and 24 fluconazole resistant
C. albicans were included to the study. Fluconazole resistant strains had more bands when evaluating with the agarose gel electrophoresis but there were no specific discriminatory band patterns to warrant the differentiation of the resistance.
The identification of
Candida species with the amplification of intergenic spacer region and RFLP analysis is a practical, short, and a reliable method when comparing to the conventional time-consuming
Candida species identification methods. The fluconazole susceptibility testing with AP-PCR seems to be a promising method but further studies must be performed for more specific results.
