A simple PCR-RFLP method for identification and differentiation of eleven Malassezia species

Abstract number: 1134_03_252

Mahzounieh M., Mirhendi H., Makimura K., Zommorodian K., Keivanlu M.

Malassezia species are part of resident skin flora of human and other warm-blooded vertebrates. These yeasts are associated with various superficial diseases including pityriasisversicolor, seborrheicdermatitis and folliculitis, as well as nosocomial blood-stream infection in paediatric care units. Although various DNA-based molecular methods have been described, a simple, reliable and cost effective method is still needed for differentiation of Malassezia species. In this study, a PCR-RFLP method using one primer pair to amplification of a 580 bp fragment related to 26S rDNA and using 2 restriction enzymes, CfoI and BstF51 was developed to clearly identify and differentiate 11 Malassezia species including M. furfur, M. pachydermatis, M. globosa, M. obtuse, M. restricta, M. sympodialis, M. dermatis, M. slooffiae, M. nana, M. japonica and M. yamatoensis. Malassezia type and standard strains were examined to verify the method. Thirteen clinical isolates were also identified in this study. The results of PCR-RFLP analysis of clinical isolates were completely comparable with those from DNA sequencing techniques. This method enables the cost effective, rapid and reliable identification of Malassezia species and therefore it could be suitable for laboratory applications.