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Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1994 Jan;38(1):45–48. doi: 10.1128/aac.38.1.45

Improved medium for fluconazole susceptibility testing of Candida albicans.

J L Rodriguez-Tudela 1, J V Martinez-Suarez 1
PMCID: PMC284394  PMID: 8141578

Abstract

We have compared fluconazole susceptibilities of 92 clinical isolates of Candida albicans by broth microdilution in two different media: standard RPMI 1640 (RPMI) and the same medium supplemented with 18 g of glucose per liter (RPMI-glucose). Preparation of media, drugs, and inocula, as well as incubation conditions, followed the preliminary recommendations of the National Committee for Clinical Laboratory Standards (Villanova, Pa.) antifungal agent working group for broth macrodilution tests with antifungal agents, adapted to microdilution. Microtiter plates were agitated for 5 min before spectrophotometric readings were performed with an automatic plate reader set at 405 mm. The MIC endpoint was defined as an inhibitory concentration calculated from the turbidimetric data as a function of the turbidity in the drug-free control wells. The mean absorbances in the drug-free wells in RPMI and RPMI-glucose were, respectively, 0.38 (41.6% transmission) and 0.99 (10.2% transmission) (P < 0.001; Student's t test). Despite the increased growth in RPMI-glucose, 98.9% of the C. albicans strains tested for fluconazole susceptibility yielded similar MICs (+/- 1 dilution) in both media. Moreover, strains with decreased susceptibility to fluconazole displaying similar MICs in both media are easier to detect in RPMI-glucose because of the greater differences between turbidimetric readings in wells with grown or fluconazole-inhibited cultures. This objective turbidimetric method, with an easy-to-read improved medium (RPMI with glucose), together with previous experience of the National Committee for Clinical Laboratory Standards antifungal agent subcommittee, could overcome some of the present problems associated with lack of reproducibility of azole susceptibility testing.

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Selected References

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  1. Anaissie E., Paetznick V., Bodey G. P. Fluconazole susceptibility testing of Candida albicans: microtiter method that is independent of inoculum size, temperature, and time of reading. Antimicrob Agents Chemother. 1991 Aug;35(8):1641–1646. doi: 10.1128/aac.35.8.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cook R. A., McIntyre K. A., Galgiani J. N. Effects of incubation temperature, inoculum size, and medium on agreement of macro- and microdilution broth susceptibility test results for yeasts. Antimicrob Agents Chemother. 1990 Aug;34(8):1542–1545. doi: 10.1128/aac.34.8.1542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dronda F., Rodríguez-Tudela J. L., Mera P., Martínez-Suárez J. Esofagitis por Candida albicans resistente a fluconazol: fracaso terapeútico y correlación in vitro. Med Clin (Barc) 1992 Jun 27;99(5):199–199. [PubMed] [Google Scholar]
  4. Espinel-Ingroff A., Kerkering T. M., Goldson P. R., Shadomy S. Comparison study of broth macrodilution and microdilution antifungal susceptibility tests. J Clin Microbiol. 1991 Jun;29(6):1089–1094. doi: 10.1128/jcm.29.6.1089-1094.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Espinel-Ingroff A., Kish C. W., Jr, Kerkering T. M., Fromtling R. A., Bartizal K., Galgiani J. N., Villareal K., Pfaller M. A., Gerarden T., Rinaldi M. G. Collaborative comparison of broth macrodilution and microdilution antifungal susceptibility tests. J Clin Microbiol. 1992 Dec;30(12):3138–3145. doi: 10.1128/jcm.30.12.3138-3145.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Galgiani J. N., Stevens D. A. Antimicrobial susceptibility testing of yeasts: a turbidimetric technique independent of inoculum size. Antimicrob Agents Chemother. 1976 Oct;10(4):721–728. doi: 10.1128/aac.10.4.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. McIntyre K. A., Galgiani J. N. In vitro susceptibilities of yeasts to a new antifungal triazole, SCH 39304: effects of test conditions and relation to in vivo efficacy. Antimicrob Agents Chemother. 1989 Jul;33(7):1095–1100. doi: 10.1128/aac.33.7.1095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Pfaller M. A., Burmeister L., Bartlett M. S., Rinaldi M. G. Multicenter evaluation of four methods of yeast inoculum preparation. J Clin Microbiol. 1988 Aug;26(8):1437–1441. doi: 10.1128/jcm.26.8.1437-1441.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Pfaller M. A., Dupont B., Kobayashi G. S., Müller J., Rinaldi M. G., Espinel-Ingroff A., Shadomy S., Troke P. F., Walsh T. J., Warnock D. W. Standardized susceptibility testing of fluconazole: an international collaborative study. Antimicrob Agents Chemother. 1992 Sep;36(9):1805–1809. doi: 10.1128/aac.36.9.1805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Pfaller M. A., Rinaldi M. G., Galgiani J. N., Bartlett M. S., Body B. A., Espinel-Ingroff A., Fromtling R. A., Hall G. S., Hughes C. E., Odds F. C. Collaborative investigation of variables in susceptibility testing of yeasts. Antimicrob Agents Chemother. 1990 Sep;34(9):1648–1654. doi: 10.1128/aac.34.9.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Radetsky M., Wheeler R. C., Roe M. H., Todd J. K. Microtiter broth dilution method for yeast susceptibility testing with validation by clinical outcome. J Clin Microbiol. 1986 Oct;24(4):600–606. doi: 10.1128/jcm.24.4.600-606.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rogers T. E., Galgiani J. N. Activity of fluconazole (UK 49,858) and ketoconazole against Candida albicans in vitro and in vivo. Antimicrob Agents Chemother. 1986 Sep;30(3):418–422. doi: 10.1128/aac.30.3.418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Warnock D. W. Azole drug resistance in Candida species. J Med Microbiol. 1992 Oct;37(4):225–226. doi: 10.1099/00222615-37-4-225. [DOI] [PubMed] [Google Scholar]

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