Ann Dermatol.  2015 Dec;27(6):715-720. 10.5021/ad.2015.27.6.715.

Antifungal Susceptibility Testing with Etest for Candida Species Isolated from Patients with Oral Candidiasis

• Song YB
• Suh MK
• Ha GY
• Kim H

• KMID: 2157448
• DOI: http://doi.org/10.5021/ad.2015.27.6.715

Abstract

BACKGROUND
The necessity of performing antifungal susceptibility tests is recently increasing because of frequent cases of oral candidiasis caused by antifungal-resistant Candida species. The Etest (BioMerieux, Marcy l'Etoile, France) is a rapid and easy-to-perform in vitro antifungal susceptibility test.
OBJECTIVE
The purpose of this study was to determine the minimal inhibitory concentrations (MICs) of antifungal agents by using the Etest for Candida species isolated from patients with oral candidiasis.
METHODS
Forty-seven clinical isolates of Candida species (39 isolates of Candida albicans, 5 isolates of C. glabrata, and 3 isolates of C. tropicalis) were tested along with a reference strain (C. albicans ATCC 90028). The MIC end points of the Etest for fluconazole, itraconazole, voriconazole, and amphotericin B susceptibility were read after the 24-hour incubation of each isolate on RPMI 1640 agar.
RESULTS
All Candida isolates were found susceptible to voriconazole and amphotericin B. However, all five isolates of C. glabrata were resistant to itraconazole, among which two isolates were also resistant to fluconazole.
CONCLUSION
This study revealed that the Etest represented a simple and efficacious method for antifungal susceptibility testing of Candida species isolated from oral candidiasis patients. Therefore, voriconazole and amphotericin B should be recommended as effective alternatives for the treatment of oral candidiasis.

Keyword

Candida; Etest; Oral candidiasis

MeSH Terms

Agar
Amphotericin B
Antifungal Agents
Candida albicans
Candida*
Candidiasis, Oral*
Fluconazole
Humans
Itraconazole
Agar
Amphotericin B
Antifungal Agents
Fluconazole
Itraconazole

Figure

• 

  Fig. 1 The Etest result of a Candida albicans isolate tested against fluconazole, itraconazole, voriconazole, and amphotericin B.

Reference

1. Kundu RV, Garg A. Yeast infections: candidiasis, tinea (pityriasis) versicolor, and Malassezia (Pityrosporum) folliculitis. In : Goldsmith LA, Katz SI, Gilchrest BA, Paller AS, Loffell DJ, Wolff K, editors. Fitzpatrick's dermatology in general medicine. 8th ed. New York: McGraw-Hill;2012. p. 2298–2307.

2. Ahn HH, Park SD, Kim KM, Park CJ, Kim HW, Kim JP, et al. Infectious skin diseases. Korean Dermatological Association. Textbook of dermatology. 6th ed. Seoul: Daehan Medical Books;2014. p. 430–437.

3. Marcos-Arias C, Eraso E, Madariaga L, Carrillo-Muñoz AJ, Quindós G. In vitro activities of new triazole antifungal agents, posaconazole and voriconazole, against oral Candida isolates from patients suffering from denture stomatitis. Mycopathologia. 2012; 173:35–46.

4. Carvalhinho S, Costa AM, Coelho AC, Martins E, Sampaio A. Susceptibilities of Candida albicans mouth isolates to antifungal agents, essentials oils and mouth rinses. Mycopathologia. 2012; 174:69–76.

5. Nweze EI, Ogbonnaya UL. Oral Candida isolates among HIV-infected subjects in Nigeria. J Microbiol Immunol Infect. 2011; 44:172–177.

6. Koga-Ito CY, Lyon JP, Resende MA. Comparison between E-test and CLSI broth microdilution method for antifungal susceptibility testing of Candida albicans oral isolates. Rev Inst Med Trop Sao Paulo. 2008; 50:7–10.

7. Enwuru CA, Ogunledun A, Idika N, Enwuru NV, Ogbonna F, Aniedobe M, et al. Fluconazole resistant opportunistic oro-pharyngeal Candida and non-Candida yeast-like isolates from HIV infected patients attending ARV clinics in Lagos, Nigeria. Afr Health Sci. 2008; 8:142–148.

8. Mareş M, Mareş M, Rusu M. Antifungal susceptibility of 95 yeast strains isolated from oral mycoses in HIV-negative and HIV-positive patients. Bacteriol Virusol Parazitol Epidemiol. 2008; 53:41–42.

9. Costa CR, de Lemos JA, Passos XS, de Araújo CR, Cohen AJ, Souza LK, et al. Species distribution and antifungal susceptibility profile of oral candida isolates from HIV-infected patients in the antiretroviral therapy era. Mycopathologia. 2006; 162:45–50.

10. Bagg J, Sweeney MP, Davies AN, Jackson MS, Brailsford S. Voriconazole susceptibility of yeasts isolated from the mouths of patients with advanced cancer. J Med Microbiol. 2005; 54:959–964.

11. Belazi M, Velegraki A, Koussidou-Eremondi T, Andreadis D, Hini S, Arsenis G, et al. Oral Candida isolates in patients undergoing radiotherapy for head and neck cancer: prevalence, azole susceptibility profiles and response to antifungal treatment. Oral Microbiol Immunol. 2004; 19:347–351.

12. Tapia C, González P, Pereira A, Pérez J, Noriega LM, Palavecino E. Antifungal susceptibility for Candida albicans isolated from AIDS patients with oropharyngeal and esophageal candidiasis: experience with Etest. Rev Med Chil. 2003; 131:515–519.

13. Davies A, Brailsford S, Broadley K, Beighton D. Resistance amongst yeasts isolated from the oral cavities of patients with advanced cancer. Palliat Med. 2002; 16:527–531.

14. Silva Mdo R, Costa MR, Miranda AT, Fernandes Ode F, Costa CR, Paula CR. Evaluation of Etest and macrodilution broth method for antifungal susceptibility testing of Candida sp strains isolated from oral cavities of AIDS patients. Rev Inst Med Trop Sao Paulo. 2002; 44:121–125.

15. Kirkpatrick WR, Revankar SG, Mcatee RK, Lopez-Ribot JL, Fothergill AW, McCarthy DI, et al. Detection of Candida dubliniensis in oropharyngeal samples from human immunodeficiency virus-infected patients in North America by primary CHROMagar candida screening and susceptibility testing of isolates. J Clin Microbiol. 1998; 36:3007–3012.

16. Lass-Flörl C, Perkhofer S, Mayr A. In vitro susceptibility testing in fungi: a global perspective on a variety of methods. Mycoses. 2010; 53:1–11.

17. Arikan S. Current status of antifungal susceptibility testing methods. Med Mycol. 2007; 45:569–587.

18. Shin JH. Antifungal drug susceptibility. Hanyang Med Rev. 2006; 26:79–85.

19. Sanitá PV, Mima EG, Pavarina AC, Jorge JH, Machado AL, Vergani CE. Susceptibility profile of a Brazilian yeast stock collection of Candida species isolated from subjects with Candida-associated denture stomatitis with or without diabetes. Oral Surg Oral Med Oral Pathol Oral Radiol. 2013; 116:562–569.

20. Negri M, Henriques M, Svidzinski TI, Paula CR, Oliveira R. Correlation between Etest, disk diffusion, and microdilution methods for antifungal susceptibility testing of Candida species from infection and colonization. J Clin Lab Anal. 2009; 23:324–330.

21. Samaranayake LP, Keung Leung W, Jin L. Oral mucosal fungal infections. Periodontol 2000. 2009; 49:39–59.

22. Lee SH, Kim SW, Bang YJ. A study on the distribution of oral candidal isolates in diabetics. Korean J Med Mycol. 2002; 7:139–148.

23. Silva S, Henriques M, Hayes A, Oliveira R, Azeredo J, Williams DW. Candida glabrata and Candida albicans co-infection of an in vitro oral epithelium. J Oral Pathol Med. 2011; 40:421–427.

24. Coco BJ, Bagg J, Cross LJ, Jose A, Cross J, Ramage G. Mixed Candida albicans and Candida glabrata populations associated with the pathogenesis of denture stomatitis. Oral Microbiol Immunol. 2008; 23:377–383.

25. Kim YJ, Suh MK, Ha GY. Azole antifungal susceptibility testing of candida species using E test. Korean J Dermatol. 2001; 39:654–659.

26. Ranque S, Lachaud L, Gari-Toussaint M, Michel-Nguyen A, Mallié M, Gaudart J, et al. Interlaboratory reproducibility of Etest amphotericin B and caspofungin yeast susceptibility testing and comparison with the CLSI method. J Clin Microbiol. 2012; 50:2305–2309.

27. Buchta V, Vejsova M, Vale-Silva LA. Comparison of disk diffusion test and Etest for voriconazole and fluconazole susceptibility testing. Folia Microbiol (Praha). 2008; 53:153–160.

28. Morace G, Borghi E, Iatta R, Amato G, Andreoni S, Brigante G, et al. Antifungal susceptibility of invasive yeast isolates in Italy: the GISIA3 study in critically ill patients. BMC Infect Dis. 2011; 11:130.

29. Pfaller MA, Andes D, Arendrup MC, Diekema DJ, Espinel-Ingroff A, Alexander BD, et al. Clinical breakpoints for voriconazole and Candida spp. revisited: review of microbiologic, molecular, pharmacodynamic, and clinical data as they pertain to the development of species-specific interpretive criteria. Diagn Microbiol Infect Dis. 2011; 70:330–343.

30. Scott LJ, Simpson D. Voriconazole: a review of its use in the management of invasive fungal infections. Drugs. 2007; 67:269–298.