Abstract: Objectives To develop a rapid genotyping method of clinical isolates of Malassezia from patients with tinea versicolor by PCR-RFLP,and to evaluate reliability of the approach as compared with biochemical classification.Methods Tween assimilation test and catalase reaction were carried out to identify 74 isolates of Malassezia species from patients with tinea versicolor and 7 Malassezia reference strains.The sequence of 28S rDNA of Malassezia species was amplified by PCR,and then the product was analyzed by RFLP with Eco88I,Bsp143Ⅱ and BshNⅠ,respectively.Results M.restricta,M.obtusa and M.pachydermatis were successfully identified by three restriction endonucleases.M.restricta was found to be more diverse from the other 6 species in genetic homology.By comparison with PCR-RFLP technique,a possible mistake was discovered with biochemical method.Conclusion PCR-RFLP is a promising molecular biological technique,which could rapidly and correctly classify Malassezia species.

Key words: Malassezia, Tinea versicolor