Heteroduplex mobility assay of the 26S rDNA D1/D2 region for differentiation of clinically relevant Candida species.
The Heteroduplex Mobility Assay (HMA) method using the PCR amplified D1/D2 region of the 26S rDNA was tested for the differentiation of clinically relevant Candida species. Strains belonging to the same species are not expected to form heteroduplexes in this assay when their PCR products are mixed. D1/D2 HMA experiments between all Candida type strains tested showed heteroduplex formation, including Candida albicans and Candida dubliniensis. There was no heteroduplex formation when most clinical and non-type strains were tested against the type strain of their presumptive species, except when C. albicans WVE and C. dubliniensis TAI were analysed. Additional HMA experiments, phenotypic characterisation, and D1/D2 sequencing identified these isolates as Candida tropicalis and Candida parapsilosis, respectively. HMA provides a rapid and relatively simple molecular tool for the differentiation of potentially pathogenic Candida species.
Ramos
Jesus P
JP
Departamento de Genética, Instituto de Biologia, CCS, bloco A, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brasil.
Rosa
Carlos A
CA
Carvalho
Elaine M M
EM
Leoncini
Orilio
O
Valente
Patricia
P
D001483:Base Sequence; D002175:Candida / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D002176:Candida albicans / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D041022:Candida tropicalis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics* / Q000302:isolation & purification; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D012331:RNA, Fungal / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D013045:Species Specificity
D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012331:RNA, Fungal; D012335:RNA, Ribosomal; C065096:RNA, ribosomal, 26S