This research was conducted on the cultured samples of 160 healthy men and women aged 0–80 years without any skin disease. Nineteen clinical isolates of Malassezia dermatis showed positive in a catalase test and all grew in 0.5% Tween-60 and 0.1% Tween-80 added to 2% glucose/1% peptone culture medium. Round and ellipsoidal yeast cells and budding of the yeast cells were observed by microscopy, resembling Malassezia sympodialis, Malassezia furfur and Malassezia nana. The 26S rDNA polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) pattern was the same as for M. dermatis (JCM 11348), the standard strain. 26S rDNA and ITS1 sequencing were performed for exact identification, showing 99% accordance with M. dermatis (AB070361), M. dermatis (AB070356), confirming the species to be new and first to be reported in Korea. Taking a molecular biological classification approach by analyzing the 26S rDNA PCR-RFLP patterns, we have successfully isolated 19 cases of M. dermatis- the first in Korea.