Estimation of minimum sterol 14alpha-demethylation-inhibitory concentration of azoles in Candida yeasts using acetate-mediated growth inhibition: potential utility in susceptibility testing.
We have recently shown that 14alpha-demethylation-deficient cells of Candida albicans are subject to growth arrest by 0.24 M acetate in a yeast extract-peptone-glucose medium and that the minimum concentration of an azole antifungal agent required for total inhibition of sterol 14alpha-demethylation (MDIC for minimum demethylation-inhibitory concentration) is practically identical to its MIC determined in the acetate-supplemented medium (O. Shimokawa and H. Nakayama, Antimicrob. Agents Chemother. 43:100-105, 1999). In the present study we estimated the MDICs of three different azoles (fluconazole, ketoconazole, and itraconazole) for strains of various Candida species using this method and compared them with the MICs determined in the corresponding acetate-free medium. The results demonstrated that the test strains were divided into two classes. One class of strains was characterized by tolerance to 14alpha-demethylation deficiency (MIC > MDIC) and consisted of strains of C. albicans, C. guilliermondii, C. kefyr, and C. tropicalis. The other class was intolerant to 14alpha-demethylation deficiency (MIC approximately MDIC) and comprised strains of C. glabrata, C. krusei, and C. parapsilosis. We also showed that replacement of the yeast extract-peptone-glucose medium with RPMI 1640 medium did not affect the results substantially. Furthermore, the 80% inhibitory concentration (IC(80)) in RPMI 1640 medium, recommended as a substitute for the conventional MIC in susceptibility testing, was found to be close to the MDIC.
Shimokawa
O
O
Department of Microbiology, Faculty of Dentistry, Kyushu University Higashi-ku, Fukuoka 812-8582, Japan. simo@dent.kyushu-u.ac.jp
Nakayama
H
H
D000085:Acetates / Q000378:metabolism* / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology*; D001393:Azoles / Q000494:pharmacology*; D002175:Candida / Q000187:drug effects* / Q000201:enzymology / Q000254:growth & development; D002177:Candidiasis / Q000382:microbiology; D003470:Culture Media; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D010088:Oxidoreductases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D058886:Sterol 14-Demethylase
D000085:Acetates; D000935:Antifungal Agents; D001393:Azoles; C548418:CYP51A1 protein, human; D003470:Culture Media; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D058886:Sterol 14-Demethylase