Evaluation of Susceptibility of Conventional Anti Fungal Drugs with Flow Cytometry Technique

Document Type : Research Paper

Authors

Abstract

Introduction: During the last decades, the incidence of fungal infection has been increased in many countries. Because of the increasing resistantce to antifungal agents, determination of an efficient strategic plan for treatment of fungal diseases is an important issue in clinical mycology.
Material and Methods:Many methods have been introduced and developed to determine invitro susceptibility tests. During the recent years, flow cytometry has been developed to solve the problem, and many papers have documented the usefulness of this technique. This study was done to evaluate the invitro susceptibility of standard PTCC strain and some clinical isolates of Candidaevaluated by macrodilution broth method according to NCCLS guidelines and flow cytometry susceptibility test. This descriptive study was done in Iran Medical University. Clinical isolates of Candidawere treated by Amphotericin B, Clotrimazole, Fluconazol, Ketoconazol and miconazole and then evaluated by flow cytometry and microdilution. Data was gathered in a questionnaire and analyzed by descriptive statistics.
Results:The data indicated that macrodilution broth methods and flow cytometry have the same results in determination of MIC for amphotericinB, clotrimazole, fluconazole, ketoconazole and miconazole in Candida albicans PTCC5027 as well as clinical candida isolate, such as C.albicans, C.kefyr, C.glabrata, C.parapsilosis and C.dubliniensis.
Conclusion:Comparing the result obtained by macrodilution broth and flow cytometry methods revealed that flow cytometry was more convenient and faster than broth methods. It is suggested that flow cytometry susceptibility test can be used as a powerful tool in determination of MIC and administration of the best antifungal drug in treatment of patients with Candida infections especially systemic candidiasis.

Keywords


1- Ramani R, Chaturvedi V. Flow cytometry antifungal susceptitility testing of pathogenic yeasts other than
candida albicans and comparison with the NCCLS broth MICrodilution test. AntiMICrob. Agent Chemother. 2000;
44: 2752 - 8.
2- Kirk S M, Callister S. M, Lim L.C.L, Schell R.F. Rapid susceptibility testing of candida albicans by flow
cytometry. J Clin-MICrobiol 1997; 35: 358-363.
3- Wenisch C, Moore C, Krause R, Presterl E Antifungal susieptibility testing of fluconazole by flow cytometry
correlates with clinical outcome. J Clin MICro July 2001; 2458-62.
4- Chaturvedi V, Ramani R Pfaller M.A.Collaborative study of the NCCLS and flow cytometry methods for
antifungal testing of candida albicans. J Clin Microbiol 2004; 42:2249-51.
5- Rudensky B, Broidie E, et al. Rapid flow-cytometric susceptibility testing of candida species.J of Antimicrobial
chemotherapy 2005;55:106-109.
6- Mitchell M, Hudspeth M, Wright A. Flow Cytometry Susceptibility Testing for the Antifungal Caspofungin J
Clin Microbiol June 2005; 2586–9.
7- Ramani R, Ramani A, Wong SJ. Rapid Flow cytometric susceptibility Testing of candida albicans. J Clin
MICrobiol 1997; 35: 2320-4.
8- Green L, Petersen B, Steimel L. Haeber P. Current W. Rapid determination of antifungal activity by flow
cytometry. J Clin MICrobiol 1994; 32: 1088-91.
9- Wenisch C, Linnau KF, Parschalk B, Zed twitz – Liebenstein K, Georgopoulos A. Rapid susceptibility testing
of fungi by flow cytometry using vital staining. J Clin MICro 1997; 35: 5-10.
10- National Committee for clinical loboratory standards. 2002. Reference Method for Broth Dilution antifungal
Susceptibility Testing of yeasts; Approved Standard second Edition.
11- اوانس ای جی، ریچارد سن ان جی. «قارچ شناسی پزشکی روشهای عملی» ترجمه خسروی، علیرضا. فصل 11. صفحة 313-349، انتشارات جهاد دانـشگاهی
دانشکده دامپزشکی دانشگاه تهران. چاپ دوم 1382.
12 - زینی، فریده. امامی، مسعود و مهبد، امیرسید علی «قارچشناسی پزشکی جامع» فصل 9 صفحة 413-455، انتشارات دانشگاه تهران، چاپ پنجم، پائیز 1376.
13- Ghannoum M A, Rex JH, Galgiani J. N. Susceptibility testing of Fungi: current status of correlation of in vitro
data with clinical outcome. J Clin MICrobiol 1996; 34: 489-495.
14- O’Gorman M R, Hopfer RL. Amphotericin B suceptibility testing of candida species by flow cytometry
Cytometry 1991; 12: 743-7.
15- Martin E, Schlasius U, Bhakdi S. Flow cytometric assay for estimating fungicidal activity of amphotericin B in
human serum. Med MIC Immun 1992; 181: 117-126.