Phenotypes and randomly amplified polymorphic DNA profiles of Candida albicans isolates from root canal infections in a Finnish population

Abstract

A total of thirty-seven Candida albicans isolates from root canal infections in a Finnish population were subtyped using phenotypic and genotypic methods. A previously described biotyping method based on the presence of five different enzymes, assimilation of eleven different carbohydrates and boric acid sensitivity of the yeasts was used to determine the phenotype. Commercially available API ZYM and API 20 C test kits were used to determine the presence of enzymes and assimilation of carbohydrates. The sensitivity of the isolates to boric acid was tested by their ability to grow on yeast-nitrogen-agar with incorporated boric acid (1.8 mg. ml(-1)). Combination of the tests revealed a total of 14 different phenotypes. The majority of the isolates, 26 strains, were classifiable into three major phenotypes: 16 isolates (43.2%) belonged to phenotype A1R, six (16.2%) to A1S and four (10.8%) to B1S. The remaining 11 phenotypes represented only a single isolate each. The randomly amplified polymorphic DNA profiles were used to determine the genotypes. For this purpose two different primers, RSD6 and RSD12 were used to develop a combination randomly amplified polymorphic DNA profile for each isolate. Altogether 31 genotypes were noted among the 37 isolates, of which only three pairs of isolates presented with congruent phenotypic and genotypic profiles. The heterogeneity of both the phenotypic and randomly amplified polymorphic DNA profiles of C. albicans isolates from root canal infections is akin to previous reports from other oral and non-oral sources in different geographic locales.

Genetic relatedness of subgingival and buccalCandida dubliniensisisolates in immunocompetent subjects assessed by RAPD-PCR

“…Such genetic heterogeneity within isolates was reported from other oral and non-oral sources in C. albicans (25, 26, 56). …”

Distribution of Candida albicans in the oral cavity of children aged 3-5 years of Uygur and Han nationality and their genotype in caries-active groups

“…The primers RSD6: 5'-GCGATCCCCA-3' (Waltimo et al, 2001) and RSD12: 5'-GCATATCAATAAGCGCAGGAAAAG-3' (Leung et al, 2000) were used for the PCR with a total volume of 30 mL that included 15 mL 2X EasyTaq PCR SuperMix and 1 mL 10 mM DNA template primer. Reaction conditions were: 94°C for 30 s, 27°C for 2 min, and 72°C for 2 min for 5 cycles, followed by 94°C for 30 s, 32°C for 2 min, and 72°C for 2 min for 45 cycles, and finally a 15-min elongation step at 72°C.…”

Fluconazole Susceptibility and Genotypic Heterogeneity of Oral Candida albicans Colonies from the Patients with Cancer Receiving Chemotherapy in China

“…Genotypic was determined using randomly amplified polymorphic DNA-PCR (RAPD) analysis. Two primers RSD6 (5'-GCG ATC CCC A-3') and RSD12 (5'-GCA TAT CAA TAA GCG CAG GAA AAG-3') for RAPD were those described by Fell and Akopyanz (Akopyanz et al, 1992;Fell, 1993 The amplification products were characterized by electrophoresis on 1.2% agarose gels in 1× TBE buffer at 50 V for 120 minutes, stained in a solution of 0.5 µg⋅mL -1 of ethidium bromide, and then visualized by UV transillumination (Bio Rad, USA) (Waltimo et al, 2001).…”