Identification of Malassezia species using direct PCR-sequencing on clinical samples from patients with pityriasis versicolor and seborrheic dermatitis

Document Type : Original Articles


1 Department of Medical Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Skin Diseases and Leishmaniasis Research Center, Department of Dermatology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran


Background and Purpose: Malassezia yeasts are lipophilic normal flora of the skin in humans and other warm-blooded vertebrates. This genus includes 18 species and is responsible for dermatological disorders, such as pityriasis versicolor, atopic dermatitis, seborrheic dermatitis, folliculitis, and dandruff. The aim of the present study was to identify the etiologic agents of Malassezia infections among the patients referring to the Referral Dermatology Clinic of Al-Zahra Hospital, Isfahan, Iran, during 2018-2019.
Materials and Methods: For the purpose of the study, clinical specimens, including skin scrapings and dandruff, were collected and subjected to direct microscopy, culture, and polymerase chain reaction (PCR) sequencing. Direct PCR was performed on the clinical samples to amplify the D1/D2 region of 26S rDNA, using specific primers; subsequently, the amplicons were sent for sequencing.
Results: This study was conducted on 120 patients with suspected pityriasis versicolor and seborrheic dermatitis, who referred to the Referral Dermatology Clinic of Al-Zahra Hospital, Isfahan, Iran, during 2018-2019. Out of this population, 50 (41.7%), 26 (52%), and 24 (48%) cases had Malassezia infection, pityriasis versicolor, and seborrheic dermatitis, respectively. Malassezia globosa was found to be the most prevalent species (n=29, 58%), followed by M. restricta (n=20, 40%), and M. arunalokei (n=1, 2%).
Conclusion: The epidemiologic study was indicative of the frequency of some Malassezia species, such as M. globosa and M. restricta, in Isfahan, Iran. It can be concluded that direct PCR on clinical samples could be used as a simple, precise, effective, fast, and affordable method for research and even routine medical mycology laboratory studies.


1. Angiolella L, Carradori S, Maccallini C, Giusiano G, Supuran TC. Targeting Malassezia species for novel synthetic and natural antidandruff agents. Curr Med Chem. 2017; 24(22):2392-412.
2. Theelen B, Cafarchia C, Gaitanis G, Bassukas ID, Boekhout T, Dawson TL Jr. Malassezia ecology, pathophysiology, and treatment. Med Mycol. 2018; 56(Suppl 1):S10-25.
3. Diongue K, Kébé O, Faye M, Samb D, Diallo M, Ndiaye M, et al. MALDI-TOF MS identification of Malassezia species isolated from patients with pityriasis versicolor at the Seafarers’ Medical Service in Dakar, Senegal. J Mycol Med. 2018; 28(4):590-3.
4. Gaitanis G, Velegraki A, Frangoulis E, Mitroussia A, Tsigonia A, Tzimogianni A, et al. Identification of Malassezia species from patient skin scales by PCR-RFLP. Clin Microbiol Infect. 2002; 8(3):162-73.
5. Zarei Mahmoudabadi A, Zarrin M, Azish M. Detection of Malassezia species isolated from patients with pityriasis versicolor and seborrheic dermatitis using Nested-PCR. Jentashapir J Health Res. 2015; 5(6):e26683.
6. Yamada Y, Makimura K, Merhendi H, Ueda K, Nishiyama Y, Yamaguchi H, et al. Comparison of different methods for extraction of mitochondrial DNA from human pathogenic yeasts. Jpn J Infect Dis. 2002; 55(4):122-5.
7. Gupta A, Kohli Y, Summerbell R. Molecular differentiation of seven Malassezia species. J Clin Microbiol. 2000; 38(5):1869-75.
8. Mirhendi H, Makimura K, Zomorodian K, Yamada T, Sugita T, Yamaguchi H. A simple PCR-RFLP method for identification and differentiation of 11 Malassezia species. J Microbiol Methods. 2005; 61(2):281-4.
9. Ayhan M, Sancak B, Karaduman A, Arikan S, Sahin S. Colonization of neonate skin by Malassezia species: relationship with neonatal cephalic pustulosis. J Am Acad Dermatol. 2007; 57(6):1012-8.
10. 1Ashbee HR. Update on the genus Malassezia. Med Mycol. 2007; 45(4):287-303.
11. Guého E, Midgley G, Guillot J. The genus Malassezia with description of four new species. Antonie Van Leeuwenhoek. 1996; 69(4):337-55.
12. Sugita T, Takashima M, Kodama M, Tsuboi R, Nishikawa A. Description of a new yeast species, Malassezia japonica, and its detection in patients with atopic dermatitis and healthy subjects. J Clin Microbiol. 2003; 41(10):4695-9.
13. Sugita T, Takashima M, Shinoda T, Suto H, Unno T, Tsuboi R, et al. New yeast species, Malassezia dermatis, isolated from patients with atopic dermatitis. J Clin Microbiol. 2002; 40(4):1363-7.
14. Sugita T, Tajima M, Takashima M, Amaya M, Saito M, Tsuboi R, et al. A new yeast, Malassezia yamatoensis, isolated from a patient with seborrheic dermatitis, and its distribution in patients and healthy subjects. Microbiol Immunol. 2004; 48(8):579-83.
15. Hirai A, Kano R, Makimura K, Duarte ER, Hamdan JS, Lachance MA, et al. Malassezia nana sp. nov., a novel lipid-dependent yeast species isolated from animals. Int J Syst Evol Microbiol. 2004; 54(Pt 2):623-7.
16. Cabañes FJ, Theelen B, Castellá G, Boekhout T. Two new lipid-dependent Malassezia species from domestic animals. FEMS Yeast Res. 2007; 7(6):1064-76.
17. Cabañes F, Vega S, Castellá G. Malassezia cuniculi sp. nov., a novel yeast species isolated from rabbit skin. Med Mycol. 2011; 49(1):40-8.
18. Paulino LC, Tseng CH, Strober BE, Blaser MJ. Molecular analysis of fungal microbiota in samples from healthy human skin and psoriatic lesions. J Clin Microbiol. 2006; 44(8):2933-41.
19. Paulino LC, Tseng CH, Blaser MJ. Analysis of Malassezia microbiota in healthy superficial human skin and in psoriatic lesions by multiplex real-time PCR. FEMS Yeast Res. 2008; 8(3):460-71.
20. Castellá G, Coutinho SD, Cabañes FJ. Phylogenetic relationships of Malassezia species based on multilocus sequence analysis. Med Mycol. 2014; 52(1):99-105.
21. Prohic A, Simic D, Sadikovic TJ, Krupalija-Fazlic M. Distribution of Malassezia species on healthy human skin in Bosnia and Herzegovina: correlation with body part, age and gender. Iran J Microbiol. 2014; 6(4):253-62.
22. Rodoplu G, Saracli M, Gümral R, Yildiran ST. Distribution of Malassezia species in patients with pityriasis versicolor in Turkey. J Mycol Med. 2014; 24(2):117-23.
23. Rasi A, Naderi R, Heshmatzade Behzadi A, Falahati M, Farehyar S, Honarbakhsh Y, et al. Malassezia yeast species isolated from Iranian patients with pityriasis versicolor in a prospective study. Mycoses. 2010; 53(4):350-5.
24. Difonzo E, Faggi E, Bassi A, Campisi E, Arunachalam M, Pini G, et al. Malassezia skin diseases in humans. G Ital Dermatol Venereol. 2013; 148(6):609-19.
25. Jagielski T, Rup E, Ziółkowska A, Roeske K, Macura AB, Bielecki J. Distribution of Malassezia species on the skin of patients with atopic dermatitis, psoriasis, and healthy volunteers assessed by conventional and molecular identification methods. BMC Dermatol. 2014; 14:3.
26. Kaur M, Narang T, Bala M, Gupte S, Aggarwal P, Manhas A. Study of the distribution of Malassezia species in patients with pityriasis versicolor and healthy individuals in Tertiary Care Hospital, Punjab. Indian J Med Microbiol. 2013; 31(3):270-4.
27. Lee Y, Yim S, Lim S, Choe Y, Ahn K. Quantitative investigation on the distribution of Malassezia species on healthy human skin in Korea. Mycoses. 2006; 49(5):405-10.
28. Guillot J, Gueho E, Lesourd M, Midgley G, Chevrier G, Dupont B. Identification of Malassezia species. A practical approach. J Mycol Med. 1996; 6(3):103-10.
29. Mayser P, Haze P, Papavassilis C, Pickel M, Gruender K, Gueho E. Differentiation of Malassezia species: selectivity of Cremophor El, castor oil and ricinoleic acid for M. furfur. Br J Dermatol. 1997; 137(2):208-13.
30. Tarazooie B, Kordbacheh P, Zaini F, Zomorodian K, Saadat F, Zeraati H, et al. Study of the distribution of Malassezia species in patients with pityriasis versicolor and healthy individuals in Tehran, Iran. BMC Dermatol. 2004; 4:5.
31. Berenji F, Mahdavi Sivaki M, Sadabadi F, Andalib Aliabadi Z, Ganjbakhsh M, Salehi M. A retrospective study of cutaneous fungal infections in patients referred to Imam Reza Hospital of Mashhad, Iran during 2000-2011. Curr Med Mycol. 2016; 2(1):20-3.
32. Honnavar P, Prasad GS, Ghosh A, Dogra S, Handa S, Rudramurthy SM. Malassezia arunalokei sp. nov., a novel yeast species isolated from seborrheic dermatitis patients and healthy individuals from India. J Clin Microbiol. 2016; 54(7):1826-34.
Volume 6, Issue 3
September 2020
Pages 21-26
  • Receive Date: 11 May 2020
  • Revise Date: 08 July 2020
  • Accept Date: 09 July 2020
  • First Publish Date: 01 September 2020