Rapid antifungal susceptibility testing of fluconazole and amphotericin B by flow cytometry using FUN-1®: a preliminary study
✍ Scribed by E. Parisi-Duchêne; C. Reibel; I. Grawey; R. Heller; I. Mazurier; D.A. de Briel; P. Moskovtchenko
- Publisher
- Masson Editeur
- Year
- 2006
- Tongue
- French
- Weight
- 557 KB
- Volume
- 16
- Category
- Article
- ISSN
- 1156-5233
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✦ Synopsis
Objective. -The increasing incidence of severe fungal infections and the difficulties encountered with the existing antifungal susceptibility testing methods led us to develop a new technique based on flow cytometry, using the fluorochrome FUN -1 ® . Materials and methods. -We tested the susceptibility of 14 Candida spp. strains (5 reference and 9 clinical strains) to amphotericin B and fluconazole by flow cytometry versus M27-A2 CLSI reference method. Fluorescence was measured with a FACScalibur ® flow cytometer (Becton Dickinson). Amphotericin B susceptibility was studied on the 14 strains whereas fluconazole one was only performed on the five reference strains.
Results. -Results were obtained after 30 min incubation for amphotericin B and 6 h for fluconazole. Reproducibility and repeatability tests have been performed on C. albicans ATCC 90028 and were > 95% and > 98%, respectively. The flow cytometry method resulted in clearcut endpoints, which abolishes the source of variability of the M27-A2 reading. The results obtained showed that the patterns of susceptibility to amphotericin B were the same with the 2 methods used for all the strains. Our preliminary study with fluconazole gave the same results with the 5 control strains. Conclusion. -FUN-1 ® staining seems to be a rapid alternative to conventional methods to assess susceptibility of Candida spp. clinical isolates.