Comparison of broth microdilution and agar dilution methods for antifungal susceptibility testing of Malassezia speciesin vitro

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XU Sheng-jing()
CAO Shuang-lin(Department of Dermatology, Affiliated Hospital of Nantong University,Nantong 226001, Jiangsu, China)
XIA Ji-ning()
SHEN Yong-nian()
L(U) Gui-xia()
LIU Wei-da()
FU Lin-ling()
Journal Title:
Chinese Journal of Dermatology
Volume 44, Issue 10, 2011
Key Word:
Malassezia;Microbial sensitivity tests;Broth microdilution method;Agar dilution method

Abstract: Objective To compare broth microdilution and agar dilution methods for in vitro testing of activities of fluconazole,ketoconazole and itraconazole against clinical Malassezia isolates.Methods Broth microdilution and agar dilution methods were used to determine the minimal inhibitory concentration(MIC)of fluconazole,ketoconazole and itraconazole for 27 clinical strains(5 species)of Malassezia.Results The minimal inhibitory concentration(MIC)ranges of fluconazole,ketoconazole and itraconazole were 0.25-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.125 mg/L respectively as shown by broth microdilution method,2-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.25 mg/L respectively as revealed by agar dilution method.Both methods demonstrated that itraconazole possessed the strongest activity against Malassezia species,followed by ketoconazole and fluconazole.The agreement rate in MICs between the two methods was 78.8%,85.2% and 88.9%,respectively for fluconazole,ketoconazole and itraconazole,with the intraclass correlation coefficients (ICCs)being 0.88,0.80 and 0.76 respectively.Conclusions Fluconazole,ketoconazole and itraconazole are highly active against Malassezia species in vitro,and itraconazole is the most active.Broth microdilution and agar dilution method coincide well in,and are applicable for,the antifungal susceptibility testing of Malassezia species in vitro.

  • [1]Gupta AK,Batra R,Bluhm R,et al.Skin diseases associated with Malassezia species.J Am Acad Dermatol,2004,51(5):785-798.
  • [2]Guého E,Midgley G,Guillot J.The genus Malassezia with description of four new species.Antonie Van Leeuwenhoek,1996,69(4):337-355.
  • [3]Guillot J,Guého E.The diversity of Malassezia yeasts confirmed by rRNA sequence and nuclear DNA comparisons.Antonie Van Leeuwenhoek,1995,67(3):297-314.
  • [4]Sugita T,Takashima M,Shinoda T,et al.New yeast species,Malassezia dermatis,isolated from patients with atopic dermatitis.J Clin Microbiol,2002,40(4):1363-1367.
  • [5]Mirhendi H,Makimura K,Zomorodian K,et al.A simple PCRRFLP method for identification and differentiation of 11 Malassezia species.J Microbiol Methods,2005,61(2):281-284.
  • [6]Hirai A,Kano R,Makimura K,et al.Malassezia nans sp.nov.,a novel lipid-dependent yeast species isolated from animals.Int J Syst Evol Microbiol,2004,54(2):623-627.
  • [7]Guillot J,Guého E,Lesourd M,et al.Identification of Malassezia species.A practical approach.J Mycol Med,1996,6(3):103-110.
  • [8]Gupta AK,Kohli Y,Li A,et al.In vitro susceptibility of the seven Malassezia species to ketoconazole,voriconazole,itraconazole and terbinafine.Br J Dermatol,2000,142(4):758-765.
  • [9]Leeming JP,Notman FH.Improved methods for isolation and enumeration of Malassezia furfurfrom human skin.J Clin Microbiol,1987,25(10):2017-2019.
  • [10]Velegraki A,Alexopoulos EC,Kritikou S,et al.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.J Clin Microbiol,2004,42(8):3589-3593.
  • [11]National Committee for Clinical Laboratory Standards(NCCLS).Reference method for broth dilution antifungal susceptibility testing of yeasts.Approved Standard-Second Edition M27-A2.Wayne,Pennsylvania:NCCLS,2002.
  • [12]National Committee for Clinical Laboratory Standards(NCCLS).Reference method for broth dilution antifungal susceptibility testing of yeasts.Approved Standard M27-A.Wayne,Pennsylvania:NCCLS,1997.
  • [13]Strippoli V,Piacentini A,D'Auria FD,et al.Antifungal activity of ketoconazole and other azoles against Malassezia furfur in vitro and in vivo.Infection,1997,25(5):303-306.
  • [14]Hammer KA,Carson CF,Riley TV.In vitro activities of ketoconazole,econazole,miconazole,and Melaleuca altemifolia(tea tree)oil against Malassezia species.Antimicrob Agents Chemother,2000,44(2):467-469.
  • [15]Garau M,Pereiro M Jr,Palacio A.In vitro susceptibilities of Malassezia species to a new triazole,albaconazole(UR-9825),and other antifungal compounds.Antimicrob Agents Chemother,2003,47(7):2342-2344.
  • [16]Ahn KJ,Ashbee HR.Determination of minimum inhibitory concentrations of several azole antifungals for Malassezia furfur.Ann Dermatol,1996,8(1):187-194.
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