Heteroduplex mobility assay of the 26S rDNA D1/D2 region for differentiation of clinically relevant Candida species |
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| Authors: | Jesus P Ramos Carlos A Rosa Elaine M M Carvalho Orilio Leoncini Patricia Valente |
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| Institution: | (1) Departamento de Genética, Instituto de Biologia, CCS, bloco A, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brasil;(2) Departamento de Microbiologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, C.P. 486, 31270-901, Brasil;(3) Departamento de Microbiologia, ICBS, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500 sala 158, 90.050-170 Porto Alegre, RS, Brasil |
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| Abstract: | 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. |
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| Keywords: | Candida albicans D1/D2 region HMA Yeast identification 26S rDNA |
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