Molecular typing of Candida spp. by random amplification of polymorphic DNA and analysis of restriction fragment length polymorphism of ribosomal DNA repeats.
Random amplification of polymorphic DNA (RAPD) using an arbitrary oligonucleotide primer (5'-CGGTGCGACG) and analysis of restriction fragment length polymorphism of ribosomal DNA (rDNA-RFLP) after digestion of genomic DNA with restriction endonuclease EcoRI were investigated as tools for genotypic delineation beyond the species level of 91 Candida clinical isolates and four reference strains including 33 Candida albicans, 19 Candida tropicalis, 22 Candida krusei and 21 Candida (Torulopsis) glabrata. Results indicated that both techniques can be useful for typing isolates of the above species, although showing a variable discriminative potential with different species. As compared to RAPD fingerprinting, the discriminative potential of rDNA-RFLP appeared to be highest for C. albicans and lowest for C. glabrata, being overall similar for C. krusei and identical for C. tropicalis. A comparative analysis of the results obtained with the two typing techniques showed that, except for C. tropicalis, they were able to provide non-redundant information, and that their use in combination could enhance the discriminative potential for delineation among C. glabrata and C. krusei isolates.
Cresti
S
S
Dipartimento di Biologia Molecolare, Università di Siena, Italy.
Posteraro
B
B
Sanguinetti
M
M
Guglielmetti
P
P
Rossolini
G M
GM
Morace
G
G
Fadda
G
G
D002175:Candida / Q000145:classification* / Q000235:genetics; D016172:DNA Fingerprinting; D004271:DNA, Fungal / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D005838:Genotype; D006801:Humans; D015336:Molecular Probe Techniques; D012150:Polymorphism, Restriction Fragment Length; D019105:Random Amplified Polymorphic DNA Technique
D004271:DNA, Fungal; D004275:DNA, Ribosomal