New PCR primer pairs specific for Candida dubliniensis and detection of the fungi from the Candida albicans clinical isolates in Japan.
The genetic patterns of Candida dubliniensis, including type strain, were analyzed by random amplified polymorphic DNA (RAPD) method in comparison with those of reference Candida albicans genotypes A, B and C strains. This RAPD pattern analysis showed that the patterns of two strains of C. dubliniensis including the type strain are very similar to the three oligo PCR primers used, but different from other C. albicans genotypes A, B and C strains with most of the PCR primers tested. The RAPD band considered to be specific for C. dubliniensis in one oligo primer was extracted and sequenced. Based on the sequence information (417-bp), two new PCR primer pairs specific for C. dubliniensis were prepared. The survey of 58 strains of Candida clinical isolates using these newly prepared PCR primer pairs identified as C. albicans in Japan suggested the presence of one strain of C. dubliniensis. The phenotypic characteristics and the sequence analysis of the variable D1/D2 domain of the large subunit (26S) ribosomal DNA of the strain confirmed that it belongs to C. dubliniensis. The usefulness and specificity of these PCR primer pairs for the identification of C. dubliniensis are discussed.
Tamura
M
M
Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, Japan.
Watanabe
K
K
Imai
T
T
Mikami
Y
Y
Nishimura
K
K
D001483:Base Sequence; D002175:Candida / Q000235:genetics / Q000302:isolation & purification*; D002176:Candida albicans / Q000235:genetics / Q000302:isolation & purification*; D002177:Candidiasis / Q000175:diagnosis; D016172:DNA Fingerprinting; D017931:DNA Primers; D004271:DNA, Fungal / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005838:Genotype; D006801:Humans; D007564:Japan; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D019105:Random Amplified Polymorphic DNA Technique; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal