Molecular Biological Identification of Malassezia Yeasts Using Pyrosequencing
نویسندگان
چکیده
BACKGROUND A Pyrosequencing assay has been used in identification of fungal species such as Candida or Aspergillus and diagnosis of pathogenic bacteria such as Helicobacter pylori but there has been no report on successful isolation and identification of Malassezia yeasts using the pyrosequencing method. OBJECTIVE Examine the applicability and plausibility of the pyrosequencing method in identification of the Malassezia species. METHODS At internal transcribed spacer (ITS) sites 1 and 2, three primers were developed using Pyrosequencing Assay Design Software (Biotage AB). Pyrosequencing was performed on 11 standard strains and 83 genomic DNA samples obtained from 66 healthy controls aged from 1 to 80. RESULTS The eleven Malassezia standard species and 83 genomic DNA samples were successfully identified using the pyrosequencing assay. CONCLUSION The pyrosequencing method is a new tool for analysis of Malassezia yeasts, and its precision and rapidity suggests its clinical applicability.
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