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Journal of Clinical Microbiology, May 2000, p. 1869-1875, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Molecular Differentiation of Seven Malassezia Species

Aditya K. Gupta,1,* Yatika Kohli,2 and Richard C. Summerbell2,3

Division of Dermatology, Department of Medicine, Sunnybrook Health Science Center and University of Toronto, Toronto,1 and Department of Clinical Microbiology, The Hospital for Sick Children, Toronto,2 Ontario, Canada, and Centraalbureau voor Schimmelcultures, Baarn, The Netherlands3

Received 18 August 1999/Returned for modification 1 December 1999/Accepted 2 February 2000

A system based on PCR and restriction endonuclease analysis was developed to distinguish the seven currently recognized Malassezia species. Seventy-eight strains, including authentic culture collection strains and routine clinical isolates, were investigated for variation in the ribosomal DNA repeat units. Two genomic regions, namely, the large subunit of the ribosomal gene and the internal transcribed spacer (ITS) region, were amplified by PCR, and products were digested with restriction endonucleases. The patterns generated were useful in identification of five out of seven Malassezia species. M. sympodialis was readily distinguishable in that its ITS region yielded a 700-bp amplified fragment, whereas the other six species yielded an 800-bp fragment. M. globosa and M. restricta were very similar in the regions studied and could be distinguished only by performing a hot start-touchdown PCR on primers for the beta -tubulin gene. Primers based on the conserved areas of the Candida cylindracea lipase gene, which were used in an attempt to amplify Malassezia lipases, yielded an amplification product after annealing at 55°C only with M. pachydermatis. This specific amplification may facilitate the rapid identification of this organism.

* Corresponding author. Mailing address: 490 Wonderland Rd. South, Suite 6, London, Ontario, Canada N6K 1L6. Phone: (519) 657-4222. Fax: (519) 657-4233. E-mail: agupta{at}execulink.com.

Journal of Clinical Microbiology, May 2000, p. 1869-1875, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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