Hafize SAV, Ayşe BARIŞ, Deniz TURAN, Fatma ÖZAKKAŞ, Rabiye ALTINBAŞ, Sümeyye ŞEN, Nuri Kiraz KİRAZ

Candida türlerinin flukonazol, vorikonazol, amfoterisin B'ye karşı duyarlılıklarının disk difüzyon ve mikrodilüsyon yöntemiyle araştırılması

Bu çalışmada hastanemizde izole edilen Candida türlerinin iki farklı yöntemle antifungal duyarlılıklarının incelenmesi amaçlanmıştır. Hastanemizde çeşitli departmanlardan gelen klinik örneklerden toplam 200 Candida türü izole edildi. Bu türler geleneksel yöntem (germ tüp oluşturma, corn-meal tween 80 besiyerinde mikroskobik görünüm) ve ticari kit API 20C (Biomerieux, France) ile tanımlandı. Bu türlerin flukonazol (FLU), vorikonazol (VORI), amfoterisin B (AMB) antifungal minimum inhibisyon konsantrasyonu (MIK) değerleri Clinical and Laboratory Standards Institute sıvı mikrodilüsyon ve disk-difüzyon (DD) yöntemine göre değerlendirildi. İzole edilen 200 Candida türü; 100 C. albicans, 25 C. glabrata, 7 C. krusei, 43 C. parapsilosis ve 25 C. tropicalis olarak tanımlandı. Bütün Candida izolatları VORI ‘ye karşı duyarlı bulundu. Yüksek direnç (%22,5) oranı referans mikrodilüsyon yöntemi ile AMB antifungaline karşı tespit edilidi. İki yöntem arasında en iyi kategorik oran sırası ile VORI (%99), FLU (%84,9) ve AMB (%72,5) olarak hesaplandı.Vorikonazol, flukonazole dirençli izolatlarda bile in vitro olarak flukonazolden daha iyi aktivite göstermiştir. Ek olarak disk difüzyon yöntemi VORI için alternatif antifungal duyarlılık yöntemi olarak değerlendirilebilir.

Anahtar Kelimeler:

Antifungal duyarlılık, Antifungal, Sıvı Mikrodilüsyon

The Investigation of the Susceptibility of Candida Species to Fluconazole, Voriconazole, and Amphotericin B by Disc Diffusion and the Microdilution Method

In this study, we aimed to investigate the antifungal susceptibility profile of Candida isolates obtained in our hospital with two methods. A total of 200 Candida isolates were obtained from samples of patients in various departments of our hospital. Identification of each strain was performed using conventional (germ tube formation, microscopic morphology in corn meal-Tween 80 agar), and commercial kit API 20C (Biomerieux, France). MIC values of fluconazole (FLU), voriconazole (VORI), and amphotericin B (AMB) were evaluated according to Clinical Laboratory Standards Institute broth microdilution and disk diffusion (DD) method. Totally isolated 200 Candida species were identified as 100 C. albicans, 25 C. glabrata, 7 C. krusei, 43 C. parapsilosis, and 25 C. tropicalis. All isolates were susceptible to VORI. High resistance rate (22,5%) was detected for AMB antifungal by reference microdilution method. Between two methods, the best categorical ratio was calculated as VORI (99%), FLU (84.9%), and AMB (72,5%) respectively. Voriconazole exhibited better activity in vitro than fluconazole, even in isolates fluconazole resistant and disk diffusion method can be considered as alternative antifungal sensitivity method for VORI.

Keywords:

Broth microdilution, Antifungal susceptibility, Antifungal,

PDF

___

[1] Quindós G .Epidemiology of candidaemia and invasive candidiasis. A changing face. Rev Iberoam Micol. 2014;31:42-8
[2] Eschenauer GA, Nguyen MH, Clancy CJ. Is Fluconazole or an Echinocandin the Agent of Choice for Candidemia. Ann Pharmacother. 2015;49:1068-74.
[3] Furlaneto MC, Rota JF, Quesada RM,et al. Species distribution and in vitro fluconazole susceptibility of clinical Candida isolates in a Brazilian tertiary-care hospital over a 3-year period. Rev Soc Bras Med Trop. 2011 ;44:595-9.
[4] Perea S, Patterson TF. Antifungal resistance in pathogenic fungi. Clin Infect Dis. 2002; 35:1073-80.
[5] Malani AN, Kerr LE, Kauffman CA. Voriconazole: How to Use This Antifungal Agent and What to Expect. Semin Respir Crit Care Med. 2015;36:786-95.
[6] Hamill RJ. Amphotericin B formulations: a comparative review of efficacy and toxicity. Drugs. 2013 ;73:919-34
[7]Clinical Laboratory Standards Institute (CLSI), 2008. Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeast; Approved Standard-Third Edition. CLSI document M27-A3.
[8] Clinical and Laboratory Standards Institute (CLSI). Reference method for antifungal disc diffusion susceptibility testing of yeasts. In Approved Guideline. (M44-A2), 2nd ed.; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2009
[9] Kiraz N, Dag I, Oz Y, Yamac M, Kiremitci A, Kasifoglu N.Correlation between broth microdilution and disk diffusion methods for antifungal susceptibility testing of caspofungin, voriconazole, amphotericin B, itraconazole and fluconazole against Candida glabrata. J Microbiol Methods.2010;82:136-40.
[10] Arendrup M, Lundgren B, Jensen I.M, Hansen BS, Frimodt-Møller N.Comparison of Etest and a tablet diffusion test with the NCCLS broth microdilution method for fluconazole and amphotericin B susceptibility testing of Candida isolates. J. Antimicrob. Chemother.2001; 47:521–526.
[11] Diekema DJ, Meser S.A, Hollis R.J, Boyken LB, Tendolkar S, Kroeger J,et al.Evaluation of Etest and disk diffusion methods compared with broth microdilution antifungal susceptibility testing of clinical isolates of Candida spp. against posaconazole, J Clin Microbiol. 2007;45:1322-4.
[12] Azevedo AC, Bizerra FC, da Matta DA, de Almeida LP, Rosas R, Colombo AL. In vitro susceptibility of a large collection of Candida Strains against fluconazole and voriconazole by using the CLSI disk diffusion assay. Mycopathologia. 2011;171:411-6.
[13] Quindós G, Sánchez-Vargas LO, Villar-Vidal M, Eraso E, Alkorta M, Hernández-Almaraz JL.Activities of fluconazole and voriconazole against bloodstream isolates of Candida glabrata and Candida krusei: a 14-year study in a Spanish tertiary medical centre. Int J Antimicrob Agents. 2008 ;31:266-71.
[14] Noake T, Kuriyama T, White PL, et al.Antifungal susceptibility of Candida species using the Clinical and Laboratory Standards Institute disk diffusionand broth microdilution methods. J Chemother. 2007;19:283-7.
[15] Arikan S. Current status of antifungal susceptibility testing methods. Med Mycol. 2007; 45:569-87.
[16] Wang H, Xiao M, Chen SC, et al. In vitro susceptibilities of yeast species to fluconazole and voriconazole as determined by the 2010 National China Hospital Invasive Fungal Surveillance Net (CHIF-NET) study. J Clin Microbiol. 2012; 50:3952-9.
[17] Fukuoka T, Johnston D A, Winslow C A, de Groot MJ, Burt C, Hitchcock CA.et al.Genetic basis for differential activities of fluconazole and voriconazole against Candida krusei. Antimicrob. Agents Chemother. 2003. 47:1213–1219.
[18] Pfaller MA, Diekema DJ, Gibbs DL, Newell VA, Meis JF, Gould IM, et al.Results from the ARTEMIS DISK Global Antifungal Surveillance Study, 1997 to 2005: an 8.5-year analysis of susceptibilities of Candidaspecies and other yeast species to fluconazole and voriconazole determined by CLSI standardized disk diffusion testing. J. Clin. Microbiol.45:1735–1745.
[19] Pfaller MA , Boyken LB, Hollis RJ, Kroeger J, Messer SA, Tendolkar S, et al.Validation of 24-Hour Fluconazole MIC Readings versus the CLSI 48-Hour Broth Microdilution Reference Method: Results from a Global Candida Antifungal Surveillance Program J Clin Microbiol. 2008 ;46:3585-90.
[20] Barry AL, Phaller MA, Rennie RP, Fuchs PC, Brown SD. Precision and accuracy of fluconazole susceptibility testing by broth microdilution, Etest, and disk diffusion methods. Antimicrob Agents Chemother. 2002 ;46:1781-4.
[21] Espinel-Ingroff A, Canton E, Gibbs D, Wang A. Correlation of Neo-Sensitabs Tablet Diffusion Assay Results on Three Different Agar Media with CLSI Broth Microdilution M27-A2 and Disk Diffusion M44-A Results for Testing Susceptibilities of Candida spp. and Cryptococcus neoformans to Amphotericin B, Caspofungin, Fluconazole, Itraconazole, and Voriconazole J Clin Microbiol. 2007;45:858-64.