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Real-Time PCR Identification of Six Malassezia Species

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Abstract

Lipophilic yeast Malassezia species is widely found on the skin surface of humans and other animals. This fungus can cause pityriasis versicolor, Malassezia folliculitis, and seborrheic dermatitis. Still now, there is a problem with species identification of Malassezia with conventional methods. We developed a real-time polymerase chain reaction (PCR) assay with multiple hybridization probes for detecting M. globosa, M. furfur, M. restricta, M. sympodialis, M. slooffiae, and M. pachydermatis. The amplification curves and specific melting peaks of the probes hybridized with real-time PCR product were used for species identifications. The assay was further evaluated on 120 samples which were performed by swabbing from 60 domestic animals (23 goats, 10 dogs, 15 cows, 3 cats, 8 rabbits, and 1 donkey) and in 70 human samples (28 patients with pityriasis versicolor, 17 breeders, and 25 control group). Fifteen M. pachydermatis were identified from animals. From human, 61 isolates were identified as M. globosa (28), M. furfur (15), M. restricta (6), M. sympodialis (8), M. slooffiae (2), and M. pachydermatis (2). Eight cases of co-detection from 6 patients and 2 breeders were revealed. Our findings show that the assay was highly effective in identifying Malassezia species. The application of multiplex real-time PCR provides a sensitive and rapid identification system for Malassezia species, which may be applied in further epidemiological surveys from clinical samples.

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References

  1. Affes M, Ben Salah S, Makni F, Sellami H, Ayadi A (2009) Molecular identification of Malassezia species isolated from dermatitis affections. Mycoses 52:251–256

    Article  CAS  PubMed  Google Scholar 

  2. Akaza N, Akamatsu H, Sasaki Y, Kishi M, Mizutani H, Sano A, Hirokawa K, Nakata S, Nishijima S, Matsunaga K (2009) Malassezia folliculitis is caused by cutaneous resident Malassezia species. Med Mycol 47:618–624

    Article  CAS  PubMed  Google Scholar 

  3. Akaza N, Akamatsu H, Sasaki Y, Takeoka S, Kishi M, Mizutani H, Sano A, Hirokawa K, Nakata S, Matsunaga K (2010) Cutaneous Malassezia microbiota in atopic dermatitis patients differ by gender and body part. Dermatology 221:253–260

    Article  PubMed  Google Scholar 

  4. Akaza N, Akamatsu H, Sasaki Y, Takeoka S, Kishi M, Mizutani H, Sano A, Hirokawa K, Nakata S, Matsunaga K (2010) Cutaneous Malassezia microbiota of healthy subjects differ by sex, body part and season. J Dermatol 37:786–792

    Article  CAS  PubMed  Google Scholar 

  5. Ashbee HR (2007) Update on the genus Malassezia. Med Mycol 45:287–303

    Article  CAS  PubMed  Google Scholar 

  6. Buommino E, Nocera FP, Parisi A, Rizzo A, Donnarumma G, Mallardo K, Fiorito F, Baroni A, and De Martino L. (2016) Correlation between genetic variability and virulence factors in clinical strains of Malassezia pachydermatis of animal origin. New Microbiol 39(3):216

    PubMed  Google Scholar 

  7. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611–622

    Article  CAS  PubMed  Google Scholar 

  8. Cabanes FJ, Theelen B, Castella G, Boekhout T (2007) Two new lipid-dependent Malassezia species from domestic animals. FEMS Yeast Res 7:1064–1076

    Article  CAS  PubMed  Google Scholar 

  9. Cabanes FJ, Vega S, Castella G (2011) Malassezia cuniculi sp. nov., a novel yeast species isolated from rabbit skin. Med Mycol 49:40–48

    Article  CAS  PubMed  Google Scholar 

  10. Chang HJ, Miller HL, Watkins N, Arduino MJ, Ashford DA, Midgley G, Aguero SM, Pinto-Powell R, von Reyn CF, Edwards W, McNeil MM, Jarvis WR (1998) An epidemic of Malassezia pachydermatis in an intensive care nursery associated with colonization of health care workers’ pet dogs. N Engl J Med 338:706–711

    Article  CAS  PubMed  Google Scholar 

  11. Ferroni A, Suarez S, Beretti JL, Dauphin B, Bille E, Meyer J, Bougnoux ME, Alanio A, Berche P, Nassif X (2010) Real-time identification of bacteria and Candida species in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 48:1542–1548

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Gemmer CM, DeAngelis YM, Theelen B, Boekhout T, Dawson TL Jr. (2002) Fast, noninvasive method for molecular detection and differentiation of Malassezia yeast species on human skin and application of the method to dandruff microbiology. J Clin Microbiol 40:3350–3357

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Giusiano G, Sosa Mde L, Rojas F, Vanacore ST, Mangiaterra M (2010) Prevalence of Malassezia species in pityriasis versicolor lesions in northeast Argentina. Rev Iberoam Micol 27:71–74

    Article  PubMed  Google Scholar 

  14. Gueho E, Midgley G, Guillot J (1996) The genus Malassezia with description of four new species. Antonie Van Leeuwenhoek 69:337–355

    Article  CAS  PubMed  Google Scholar 

  15. Gupta AK, Boekhout T, Theelen B, Summerbell R, Batra R (2004) Identification and typing of Malassezia species by amplified fragment length polymorphism and sequence analyses of the internal transcribed spacer and large-subunit regions of ribosomal DNA. J Clin Microbiol 42:4253–4260

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Gupta AK, Kohli Y, Summerbell RC (2000) Molecular differentiation of seven Malassezia species. J Clin Microbiol 38:1869–1875

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Hirai A, Kano R, Makimura K, Duarte ER, Hamdan JS, Lachance MA, Yamaguchi H, Hasegawa A (2004) Malassezia nana sp. nov., a novel lipid-dependent yeast species isolated from animals. Int J Syst Evol Microbiol 54:623–627

    Article  CAS  PubMed  Google Scholar 

  18. Hiruma M, Cho O, Hiruma M, Kurakado S, Sugita T, Ikeda S (2014) Genotype analyses of human commensal scalp fungi, Malassezia globosa, and Malassezia restricta on the scalps of patients with dandruff and healthy subjects. Mycopathologia 177:263–269

    Article  CAS  PubMed  Google Scholar 

  19. Kim JY, Hahn HJ, Choe YB, Lee YW, Ahn KJ, Moon KC (2013) Molecular biological identification of Malassezia yeasts using pyrosequencing. Ann Dermatol 25:73–79

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Klingspor L, Jalal S (2006) Molecular detection and identification of Candida and Aspergillus spp. from clinical samples using real-time PCR. Clin Microbiol Infect 12:745–753

    Article  CAS  PubMed  Google Scholar 

  21. Kolecka A, Khayhan K, Arabatzis M, Velegraki A, Kostrzewa M, Andersson A, Scheynius A, Cafarchia C, Iatta R, Montagna MT, Youngchim S, Cabanes FJ, Hoopman P, Kraak B, Groenewald M, Boekhout T. Efficient identification of Malassezia yeasts by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Br J Dermatol 170:332–341

  22. Lee YW, Byun HJ, Kim BJ, Kim DH, Lim YY, Lee JW, Kim MN, Kim D, Chun YJ, Mun SK, Kim CW, Kim SE, Hwang JS (2011) Distribution of Malassezia species on the scalp in korean seborrheic dermatitis patients. Ann Dermatol 23:156–161

    Article  PubMed  PubMed Central  Google Scholar 

  23. Machado ML, Cafarchia C, Otranto D, Ferreira RR, Bianchi SP, Latrofa MS, Parisi A, Ferreiro L (2010) Genetic variability and phospholipase production of Malassezia pachydermatis isolated from dogs with diverse grades of skin lesions. Med Mycol 48:889–892

    Article  CAS  PubMed  Google Scholar 

  24. Makimura K, Tamura Y, Kudo M, Uchida K, Saito H, Yamaguchi H (2000) Species identification and strain typing of Malassezia species stock strains and clinical isolates based on the DNA sequences of nuclear ribosomal internal transcribed spacer 1 regions. J Med Microbiol 49:29–35

    Article  CAS  PubMed  Google Scholar 

  25. Mirhendi H, Makimura K, Zomorodian K, Yamada T, Sugita T, Yamaguchi H (2005) A simple PCR-RFLP method for identification and differentiation of 11 Malassezia species. J Microbiol Methods 61:281–284

    Article  CAS  PubMed  Google Scholar 

  26. Morishita N, Sei Y, Sugita T (2006) Molecular analysis of Malassezia microflora from patients with pityriasis versicolor. Mycopathologia 161:61–65

    Article  CAS  PubMed  Google Scholar 

  27. Paulino LC, Tseng CH, Blaser MJ (2008) Analysis of Malassezia microbiota in healthy superficial human skin and in psoriatic lesions by multiplex real-time PCR. FEMS Yeast Res 8:460–471

    Article  CAS  PubMed  Google Scholar 

  28. Prohic A, Kasumagic-Halilovic E (2009) Identification of Malassezia pachydermatis from healthy and diseased human skin. Med Arh 63:317–319

    PubMed  Google Scholar 

  29. Saad M, Sugita T, Saeed H, Ahmed A (2013) Molecular epidemiology of Malassezia globosa and Malassezia restricta in Sudanese patients with pityriasis versicolor. Mycopathologia 175:69–74

    Article  CAS  PubMed  Google Scholar 

  30. Schabereiter-Gurtner C, Selitsch B, Rotter ML, Hirschl AM, Willinger B (2007) Development of novel real-time PCR assays for detection and differentiation of eleven medically important Aspergillus and Candida species in clinical specimens. J Clin Microbiol 45:906–914

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Senczek D, Siesenop U, Bohm KH (1999) Characterization of Malassezia species by means of phenotypic characteristics and detection of electrophoretic karyotypes by pulsed-field gel electrophoresis (PFGE). Mycoses 42:409–414

    Article  CAS  PubMed  Google Scholar 

  32. Shokohi T, Afshar P, Barzgar A (2009) Distribution of Malassezia species in patients with pityriasis versicolor in Northern Iran. Indian J Med Microbiol 27:321–324

    Article  CAS  PubMed  Google Scholar 

  33. Sugita T, Kodama M, Saito M, Ito T, Kato Y, Tsuboi R, Nishikawa A (2003) Sequence diversity of the intergenic spacer region of the rRNA gene of Malassezia globosa colonizing the skin of patients with atopic dermatitis and healthy individuals. J Clin Microbiol 41:3022–3027

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Sugita T, Suzuki M, Goto S, Nishikawa A, Hiruma M, Yamazaki T, Makimura K (2010) Quantitative analysis of the cutaneous Malassezia microbiota in 770 healthy Japanese by age and gender using a real-time PCR assay. Med Mycol 48:229–233

    Article  CAS  PubMed  Google Scholar 

  35. Sugita T, Tajima M, Takashima M, Amaya M, Saito M, Tsuboi R, Nishikawa A (2004) A new yeast, Malassezia yamatoensis, isolated from a patient with seborrheic dermatitis, and its distribution in patients and healthy subjects. Microbiol Immunol 48:579–583

    Article  CAS  PubMed  Google Scholar 

  36. Sugita T, Tajima M, Tsubuku H, Tsuboi R, Nishikawa A (2006) Quantitative analysis of cutaneous Malassezia in atopic dermatitis patients using real-time PCR. Microbiol Immunol 50:549–552

    Article  CAS  PubMed  Google Scholar 

  37. Sugita T, Takashima M, Kodama M, Tsuboi R, Nishikawa A (2003) Description of a new yeast species, Malassezia japonica, and its detection in patients with atopic dermatitis and healthy subjects. J Clin Microbiol 41:4695–4699

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Sugita T, Takashima M, Shinoda T, Suto H, Unno T, Tsuboi R, Ogawa H, Nishikawa A (2002) New yeast species, Malassezia dermatis, isolated from patients with atopic dermatitis. J Clin Microbiol 40:1363–1367

    Article  PubMed  PubMed Central  Google Scholar 

  39. Sugita T, Zhang E, Tanaka T, Nishikawa A, Tajima M, Tsuboi R (2013) Recent advances in research on Malassezia microbiota in humans. Med Mycol J 54:39–44

    Article  PubMed  Google Scholar 

  40. Untergasser A, Cutcutache I, Koressaar T, Ye J, Faircloth BC, Remm M, Rozen SG (2012) Primer3–new capabilities and interfaces. Nucleic Acids Res 40:e115

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Vuran E, Karaarslan A, Karasartova D, Turegun B, Sahin F (2014) Identification of Malassezia species from pityriasis versicolor lesions with a new multiplex PCR method. Mycopathologia 177:41–49

    Article  CAS  PubMed  Google Scholar 

  42. Xie Z, Ran Y, Zhang H, Zhang M, Wan H, Li C (2014) An analysis of the Malassezia species distribution in the skin of patients with pityriasis versicolor in Chengdu, China. Sci World J 2014:182596

    Google Scholar 

  43. Zhang H, Ran Y, Xie Z, Zhang R (2013) Identification of Malassezia species in patients with seborrheic dermatitis in China. Mycopathologia 175:83–89

    Article  PubMed  Google Scholar 

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Acknowledgements

Our work was supported by the Ministry of Education and Scientific Research and the Ministry of Health.

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Authors and Affiliations

  1. Fungal and Parasitic Molecular Biology Laboratory, Faculty of Medicine, University of Sfax, Rue Magida Boulila, Sfax, 3029, Tunisia

    Amin Ilahi, Inès Hadrich, Sourour Neji, Houaida Trabelsi, Fattouma Makni & Ali Ayadi

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  1. Amin Ilahi
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  2. Inès Hadrich
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Correspondence to Inès Hadrich.

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Ilahi, A., Hadrich, I., Neji, S. et al. Real-Time PCR Identification of Six Malassezia Species. Curr Microbiol 74, 671–677 (2017). https://doi.org/10.1007/s00284-017-1237-7

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  • DOI: https://doi.org/10.1007/s00284-017-1237-7

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