Amfoterisin B, flukonazol ve vorikonazollünün kan kültürlerinden izole edilen Candida türlerine karşı antifungal aktivitesinin mikrodilüsyon ve disk diffüzyon yöntemleri ile belirlenmesi

Öz

Amaç: Candida türlerinin neden olduğu invaziv fungal enfeksiyonlar önemli ölçüde artmıştır. Kandidemiler sadece mortalite ile ilişkili olmayıp, aynı zamanda hastanede kalış süresinin uzamasına ve tıbbi bakım maliyetinin artmasına da sebep olmaktadır. Candida albicans ve non-albicans Candida türlerinin neden olduğu kandidemilerde, hastaların klinik bulgularının aynı olmasına rağmen bu türlerin antifungal ilaçlara duyarlılıkları da farklıdır. Bu çalışmada; Candida kan izolatlarının antifungal duyarlılık verilerini sunmaktadır.Yöntem: 35 Candida albicans, 25 Candida tropicalis, 15 Candida parapsilosis, 8 Candida glabrata, 4 Candida krusei ve 3 Candida kefyr olmak üzere toplam 90 Candida izolatı incelendi. Bu türlerin amfoterisin B, flukonazol ve vorikonazole karşı antifungal duyarlılıkları %2 glukozla zenginleştirilmiş RPMI 1640 agar besiyeri kullanılarak mikrodilüsyon yöntemi CLSI M27-A3 ve 0,5 µg/mL metilen mavisi ve %2 glukozla zenginleştirilmiş MuellerHinton agar besiyeri kullanılarak disk diffüzyon CLSI M44-A2 yöntemi ile belirlendi. Bulgular:Candida izolatları mikrodilüsyon yöntemi ile %87,7 oranında, disk diffüzyon yöntemi ile %82,2 oranında flukonazole duyarlı bulundu. Mikrodilüsyon yöntemi ile bu izolatların %6,6’sı %8,5 C. albicans, %4,0 C. tropicalis ve % 25,0 C. glabrata flukonazole karşı doza bağlı duyarlı, %5,5 ise dirençli olarak belirlendi. Dirençli suşların dördü C. krusei %100 , biri ise C. albicans %2,8 idi. Disk diffüzyon yöntemi ile Candida suşlarının %12,2’si doza bağlı duyarlı %17,1 C. albicans, %12,0 C. tropicalis and %25,0 C. glabrata olarak, %5,5’i [bir izolat C. albicans %2,8 , dört izolat C. krusei %100 ] ise flukonazole dirençli olarak belirlendi. Bütün suşlar amfoterisin B ve vorikonazole duyarlı olarak bulundu.Sonuç: Amfoterisin B ve vorikonazollün Candida izolatlarına olduğu artmaktadır ki bundan dolayı fungal patojenlerin duyarlılık profillerini değerlendirmek giderek önem kazanmaktadır.karşı belirlendi. in-vitro aktivitesinin yüksek direnç Antifungal ilaçlara

Anahtar Kelimeler

• Antifungal duyarlılık, flukonazol, kan izolatları

Activities of amphotericin B, fluconazole and voriconazole against Candida bloodstream isolates determined by broth microdilution and disk diffusion methods

Abstract

Objective: Invasive fungal infections caused by Candida species have increased significantly. Candidemia is not only associated with a mortality, but also extends the duration of hospital stay and increases the cost for medical care. Although, the clinical presentations of the patients with candidemia caused by Candida albicans and non-albicans Candida species are indistinguishable, the susceptibilities to antifungal agents of these species are different. This study presents data on antifungal susceptibility profiles of Candida bloodstream isolates.Method: We tested a total of 90 strains, including 35 strains of Candida albicans, 25 strains of Candida tropicalis, 15 strains of Candida parapsilosis, 8 strains of Candida glabrata, 4 strains of Candida krusei and 3 strains of Candida kefyr. Susceptibility to amphotericin B, fluconazole and voriconazole was determined by Clinical Laboratory Standards Institute broth microdilution method CLSI M27-A3 using RPMI 1640 as test medium supplemented with 2% glucose and disk diffusion methods were performed according to CLSI M44-A2 using methylene blue 0.5 µg/mL and glucose 2% enriched Mueller-Hinton agar. Results: In this study, 87.7%, 82.2% of Candida isolates were susceptible to fluconazole with the broth Mikrodilüsyon yöntemi ile bu izolatların %6,6’sı %8,5 C. albicans, %4,0 C. tropicalis ve % 25,0 C. glabrata flukonazole karşı doza bağlı duyarlı, %5,5 ise dirençli olarak belirlendi. Dirençli suşların dördü C. krusei %100 , biri ise C. albicans %2,8 idi. Disk diffüzyon yöntemi ile Candida suşlarının %12,2’si doza bağlı duyarlı %17,1 C. albicans, %12,0 C. tropicalis and %25,0 C. glabrata olarak, %5,5’i [bir izolat C. albicans %2,8 , dört izolat C. krusei %100 ] ise flukonazole dirençli olarak belirlendi. Bütün suşlar amfoterisin B ve vorikonazole duyarlı olarak bulundu.Sonuç: Amfoterisin B ve vorikonazollün Candida izolatlarına olduğu artmaktadır ki bundan dolayı fungal patojenlerin duyarlılık profillerini değerlendirmek giderek önem kazanmaktadır.karşı belirlendi. in-vitro aktivitesinin yüksek direnç Antifungal ilaçlara

Keywords

• Antifungal susceptibility, fluconazole, bloodstream isolates

Kaynakça

1. 1. Godoy P, Tiraboschi IN, Severo LC, Bustamante B, Calvo B, de Almeda LP, et al. Species distribution and antifungal susceptibility profile of Candida spp. bloodstream isolates from Latin America Hospitals. Mem Inst Oswaldo Cruz, 2003; 98 (3): 401-5.
2. 2. da Matta DA, de Almeida LP, Machado AM, Azevedo AC, Kusano EJ, Travassos NF, et al. Antifungal susceptibility of 1000 Candida bloodstream isolates to 5 antifungal drugs: results of a multicenter study conducted in Sao Paulo, Brazil, 1995-2003. Diagn Microbiol Infect Dis, 2007; 57 (4): 399-404.
3. 3. Al-Jasser AM, Elkhizzi NA. Distribution of Candida species among bloodstream isolates. Saudi Med J, 2004; 25 (5): 566-9.
4. 4. Peman J, Canton E, Gobernado M. Epidemiology and antifungal susceptibility of Candida species isolated from blood: results of a 2-year multicenter study in Spain. Eur J Clin Microbiol Infect Dis, 2005; 24: 23-30.
5. 5. Cheng MF, Yang YL, Yao TJ, Lin JY, Liu JS, Tang RB, et al. Risk factors for fatal candidemia caused by Candida albicans and non-albicans Candida species. BMC Infect Dis, 2005; 5: 22.
6. 6. Oberoi JK, Wattal C, Goel N, Raveendran R, Datta S, Prasad K. Non-albicans Candida species in bloodstream infections in a tertiary care hospital at New Delhi, India. Indian J Med Res, 2012; 136 (6): 997-1003.
7. 7. Yang YL, Wang AH, Wang CW, Cheng WT, Li SY, Lo HJ, et al. Susceptibilities to amphotericin B and fluconazole of Candida species in Taiwan surveillance of antimicrobial resistance of yeasts 2006. Diagn Microbiol Infect Dis, 2008; 61 (2): 175-80.
8. 8. Nakamura T, Takahashi H. Epidemiological study of Candida infections in blood: susceptibilities of Candida spp. to antifungal agents, and clinical features associated with the candidemia. J Infect Chemother, 2006; 12 (3): 132-8.

9. 9. Badiee P, Alborzi A. Susceptibility of clinical Candida species isolates to antifungal agents by E-test, Southern Iran: A five year study. Iran J Microbiol, 2011; 3 (4): 183-8.
10. 10. Azevedo AC, Bizerra FC, de 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 (6): 411-6.
11. 11. Moracea G, Polonelli L. Voriconazole activity against clinical yeast isolates: a multicentre Italian study. Int J Antimicrob Ag, 2005; 26: 247-53.
12. 12. Anonmyous. References method for broth dilution antifungal susceptibility testing of yeasts: USA: Clinical and Laboratory Standards Institute, Wayne, PA. Approved Standard, 3rd ed., CLSI document M27-A3, 2008.
13. 13. Anonmyous. Method for antifungal disk diffusion susceptibility testing of yeasts: USA: Clinical and Laboratory Standards Institute, Wayne, PA. Approved Guideline-2nd ed., M44-A2, 2009.
14. 14. Negri M, Henriques M, Svidzinski TIE, Paula CR, Oliveira R. Correlation between E-test disk diffusion, and microdilution methods for antifungal susceptibility testing of Candida species from infection and colonization. J Clin Lab Anal, 2009; 23: 324-30.
15. 15. Mandras N, Tullio V, Allizond V, Scalas D, Branche G, Roana J, et al. In vitro activities of fluconazole and voriconazole against clinical isolates of Candida spp. determined by disk diffusion testing in Turin, Italy. Antimicrob Agents Chemother. 2009; 53 (4): 1657-59.
16. 16. Pfaller MA, Diekema DJ, Gibbs DL, Newell VA, Ellis D, Tullio V, et al. Results from the ARTEMIS DISK global antifungal surveillance study, 1997 to 2007: a 10.5 year analysis of susceptibilities of Candida species to fluconazole and voriconazole as determined by CLSI standardized disk diffusion. J Clin Microbiol, 2010; 48 (4): 1366-77.
17. 17. Swinne D, Watelle M, Flaes MV, Nolard N. In vitro activities of voriconazole (UK-109, 496), fluconazole, itraconazole and amphotericin B against 132 non-albicans bloodstream yeast isolates (CANARI study). Mycoses, 2004; 47: 177-83.
18. 18. Gonzalez GM, Elizondo M, Ayala J. Trends in species distribution and susceptibility of bloodstream isolates of Candida collected in Monterrey, Mexico, to seven antifungal agents: results of a 3-year (2004 to 2007) surveillance study. J Clin Microbiol, 2008; 40(9): 2902-5.
19. 19. Hajjeh RA, Sofair AN, Harrison LH, Lyon GM, Arthington-Skaggs BA, Mirza SA, et al. Incidence of bloodstream infections due to Candida species and in vitro susceptibilities of isolates collected from 1998 to 2000 in a population-based active surveillance program. J Clin Microbiol, 2004; 42 (4): 1519-27.
20. 20. Takakura S, Fujihara N, Saito T, Kudo T, Linuma Y, Lchiyama S, et al. National surveillance of species distribution in blood isolates of Candida species in Japan and their susceptibility to six antifungal agents including voriconazole and micafungin. J Antimicrob Chemother, 2004; 53: 283-9.
21. 21. Mokaddas EM, Al-Sweih NA, Khan ZU. Species distribution and antifungal susceptibility of Candida bloodstream isolates in Kuwait: a 10-year study. J Med Microbiol, 2007; 56: 255-9.
22. 22. Samra Z, Yardeni M, Peled N, Bishara J. Species distribution and antifungal susceptibility of Candida bloodstream isolates in a tertiary medical in Israel. Eur J Clin Microbiol Infect Dis, 2005; 24; 592-5.
23. 23. Martin D, Perasat F, Piens MA, Picot S. Candida species distribution in bloodstream cultures in Lyon, France, 1998-2001. Eur J Clin Microbiol Infect Dis, 2005; 24; 329-33.
24. 24. Pfaller MA, Boyken L, Hollis RJ, Kroeger J, Messer SA, Tendolkar S, et al. Comparison of results of flukonazole and voriconazole disk diffusion testing for Candida spp. with results from a central reference laboratory in the ARTEMIS DISK global antifungal surveillance program. Diag Microbiol Infect Dis, 2009; 65: 27-34.
25. 25. Doczi I, Dosa E, Hajdu E, Nagy E. Aetiology and antifungal susceptibility of yeast bloodstream infections in a Hungarian University hospital between 1996 and 2000. J Med Microbiol, 2002; 51: 677-81.
26. 26. Lee MK, Kim HR, Kang JO, Kim MN, Kim EC, Kim JS, et al. Susceptibility and trailing growth of Candida albicans to fluconazole: results of a Korean multicenter study. Mycoses, 2007; 50: 148-9.
27. 27. Mallie M, Bastide JM, Blancard A, Bonnin A, Bretagne S, Cambon M, et al. In vitro susceptibility testing of Candida and Aspergillus spp. to voriconazole and other antifungal agents using E-test®: results of a French multicentre study. Int J Antimicrob Ag, 2005; 25 (4): 321-28.
28. 28. Messer SA, Jones RN, Fritsche TR. International surveillance of Candida spp. and Aspergillus spp: report from SENTRY antimicrobial surveillance program (2003). J Clin Microbiol, 2006; 44 (5): 1782-87.
29. 29. Pfaller MA, Diekema DJ. Twelve years of fluconazole in clinical practice: global trends in species distribution and fluconazole susceptibility of bloodstream isolates of Candida group. Clin Microbiol Infect, 2004; 10 (1): 11–23.
30. 30. Cuenca-Estrella M, Rodriguez D, Almirante B, Morgan J, Planes AM, Almeda M, et al. In vitro susceptibilities of bloodstream isolates of Candida species to six antifungal agents: results from a population-based active surveillance programme, Barcelona, Spain, 2002-2003. J Antimicrob Chemother, 2005; 55: 194-99.
31. 31. Kibbler CC, Seaton S, Barnes RA, Gransden WR, Holliman RE, Johnson EM, et al. Management and outcome of bloodstream infections due to Candida species in England and Wales. J Hospital Infect, 2003; 54: 18-24.