The Development and Evaluation of Multiplex PCR Technique for Identification of Malassezia Yeast

Kim, Hyeong Mi; Lim, Yun Young; Park, Eun Joo; Chun, Young Jin; Kim, Myeung Nam; Kim, Beom joon; Lee, Dong Hoon

Abstract

BACKGROUND
Malassezia yeasts as major pathogenic fungi causes the common skin diseases including dandruff, psoriasis, seborrheic dermatitis and atopic dermatitis etc. various molecular techniques were developed to identify and classify the Malassezia species until now. But, these methods were discovered the problems. So, the development of the better molecular methods required to identify and classify of Malasseiza species.
OBJECTIVE
We sought to develop of molecular techniques to identify and classify of six Malassezia species (M. restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa).
METHODS
We designed primers about ITS1 (Internal transcribed space 1) region that were well-known region useful to identify of Malassezia species. Because, ITS1 region that is located between 18S and 5.8S rDNA of ribosomal DNA was comparatively mutated quickly. The mono PCR using ITS1 primers was performed to confirm the specificity of ITS1 primers with six Malassezia standard strains. Then, Malassezia Multiplex detection kit was developed on the basis of technique using ITS1 regions. Malassezia Multiplex detection kit was used to perform multiplex PCR with six Malassezia standard strains and clinical isolates.
RESULTS
The results of mono PCR using ITS1 primers about six Malassezia standard strains was detected each Malassezia standard strains. Also, the multiplex PCR using developed Malassezia Multiplex detection kit was confirmed to classify about six Malassezia standard strains and clinical isolates.
CONCLUSION
In this study, we verified that six Malassezia yeasts was classified using Malassezia Multiplex detection kit from Malasszia standard strains and clinical isolates. And we anticipate that Malassezia Multiplex detection kit is able to do accurate diagnosis about six Malassezia yeasts (M. restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa).