| Object To investigate the distribution of Malassezia species in normal human cerumen and analyze the concordance of Malassezia species in two sides' external auditory canals from the same host. To construct cleavage map of Malassezia species by using restriction enzyme and search for a fast and convenient identified method.Methods The cerumen, came from 88 samples of 44 volunteers, were inoculated on medium containing rapeseed oil. Contrasted with 11 standard strains, the isolates were identified to species through physiological and morphological characteristics. Primers were designed to target toward 26S rDNA and genomic DNA was extracted by modified benzyl chloride complex method and followed by DNA sequence analysis of the PCR products. Use DNAssist2.0 software to design digestion pattern of these species and elect enzyme to digest.Results 1. Malassezia species were isolated from 44 of 45 individuals studied (97.78%) in there two sides' external auditory canals. Five species were identified from 88 isolated strains, including 29 strains (32.9%) of M. furfur, 23 strains (26.14%) of M. slooffiae, 18 strains (20.45%) of M. sympodialis, 11 strains (12.50%) of M. globosa and 7 strains (7.97%) of M. restricta; 2. Among 44 volunteers, isolates of Malassezia from each individual of 38 volunteers shared the same species (86.36%); 3. Modified benzyl chloride complex method can extract Malassezia genomic DNA. 4. One restriction enzyme (CfoI) can distinguish these five species with different special patterns.Conclusion The species of Malassezia is variegated in cerumen of human and the dominant species is M. furfur. In two sides' cerumen from the same host, the species of Malassezia tend to identity. Modified benzyl chloride complex method is a fit method for the extraction ofMalassezia genomic DNA. PCR-RFLP can be used to identify Malassezia species as a fast and convenient molecular method.
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