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Journal of Clinical Microbiology, August 2004, p. 3589-3593, Vol. 42, No. 8
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.8.3589-3593.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Use of Fatty Acid RPMI 1640 Media for Testing Susceptibilities of Eight Malassezia Species to the New Triazole Posaconazole and to Six Established Antifungal Agents by a Modified NCCLS M27-A2 Microdilution Method and Etest

Aristea Velegraki,^* Evangelos C. Alexopoulos, Stavroula Kritikou, and George Gaitanis

Mycology Reference Laboratory (Hellenic Centre for Diseases Control), Department of Microbiology, Medical School, University of Athens, Athens, Greece

Received 30 January 2004/ Returned for modification 28 March 2004/ Accepted 20 April 2004

A novel formulation of RPMI 1640 medium for susceptibility testing of Malassezia yeasts by broth microdilution (BMD) and Etest is proposed. A modification of the NCCLS M27-A2 BMD method was used to test 53 isolates of Malassezia furfur (12 isolates), M. sympodialis (8 isolates), M. slooffiae (4 isolates), M. globosa (22 isolates), M. obtusa (2 isolates), M. restricta (2 isolates), M. pachydermatis (1 isolates), and M. dermatis (2 isolates) against amphotericin B, ketoconazole, itraconazole, fluconazole, voriconazole, terbinafine, and posaconazole by BMD and Etest. RPMI and antibiotic medium 3 (AM3) were supplemented with glucose, bile salts, a mixture of fatty acids, and n-octadecanoate fatty acids and Tween 20. M. furfur ATCC 14521 and M. globosa ATCC 96807 were used as quality control strains. Depending on the species, MICs were read after 48 or 72 h of incubation at 32°C. Low azole and terbinafine MICs were recorded for all Malassezia species, whereas amphotericin B displayed higher MICs (16 µg/ml) against M. furfur, M. restricta, M. globosa, and M. slooffiae strains, which were AM3 confirmed. Agreement of the two methods was 84 to 97%, and intraclass correlation coefficients were statistically significant (P < 0.001). Because of higher amphotericin B MICs provided by Etest for strains also displaying high BMD MICs (1 µg/ml), agreement was poorer. The proposed media are used for the first time and can support optimum growth of eight Malassezia species for recording concordant BMD and Etest MICs.

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* Corresponding author. Mailing address: Mycology Reference Laboratory, Department of Microbiology, Medical School, University of Athens, Mikras Asias 75-77, Goudi, Athens 115 27, Greece. Phone: 30210 746 2146. Fax: 30210 746 2147. E-mail: aveleg{at}med.uoa.gr.

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Journal of Clinical Microbiology, August 2004, p. 3589-3593, Vol. 42, No. 8
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.8.3589-3593.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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