Sensitive bioassay for determination of fluconazole concentrations in plasma using a Candida albicans mutant hypersusceptible to azoles.

Details

Serval ID
serval:BIB_C57403C1A536
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Sensitive bioassay for determination of fluconazole concentrations in plasma using a Candida albicans mutant hypersusceptible to azoles.
Journal
Antimicrobial Agents and Chemotherapy
Author(s)
Marchetti O., Majcherczyk P.A., Glauser M.P., Bille J., Moreillon P., Sanglard D.
ISSN
0066-4804 (Print)
ISSN-L
0066-4804
Publication state
Published
Issued date
2001
Volume
45
Number
3
Pages
696-700
Language
english
Abstract
The antifungal agent fluconazole (FLC) is widely used in clinical practice. Monitoring FLC levels is useful in complicated clinical settings and in experimental infection models. A bioassay using Candida pseudotropicalis, a simple and cost-effective method, is validated only for FLC levels ranging from 5 to 40 mg/liter. An extension of the analytical range is needed to cover most yeast MICs. A new bioassay in RPMI agar containing methylene blue was developed using C. albicans DSY1024, a mutant rendered hypersusceptible to FLC constructed by the deletion of the multidrug efflux transporter genes CDR1, CDR2, CaMDR1, and FLU1. Reproducible standard curves were obtained with FLC concentrations in plasma ranging from 1 to 100 mg/liter (quadratic regression coefficient > 0.997). The absolute sensitivity was 0.026 microg of FLC. The method was internally validated according to current guidelines for analytical method validation. Both accuracy and precision lied in the required +/-15% range. FLC levels measured by bioassay and by high-performance liquid chromatography (HPLC) performed with 62 plasma samples from humans and rats showed a strong correlation (coefficients, 0.979 and 0.995, respectively; percent deviations of bioassay from HPLC values, 0.44% +/- 15.31% and 2.66% +/- 7.54%, respectively). In summary, this newly developed bioassay is sensitive, simple, rapid, and inexpensive. It allows nonspecialized laboratories to determine FLC levels in plasma to within the clinically relevant concentration range and represents a useful tool for experimental treatment models.
Keywords
Animals, Antifungal Agents/blood, Antifungal Agents/pharmacology, Azoles/blood, Azoles/pharmacology, Biological Assay, Candida albicans/drug effects, Candida albicans/genetics, Chromatography, High Pressure Liquid, Colony Count, Microbial, Female, Fluconazole/blood, Fluconazole/pharmacology, Humans, Microbial Sensitivity Tests, Models, Animal, Rats, Rats, Wistar
Pubmed
Web of science
Open Access
Yes
Create date
24/01/2008 13:54
Last modification date
20/08/2019 15:41
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