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Comparative Analysis of the Vitek 2 Antifungal Susceptibility System and E-test with the CLSI M27-A3 Broth Microdilution Method for Susceptibility Testing of Indian Clinical Isolates of Cryptococcus neoformans

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Abstract

The emergence of antifungal resistance among Cryptococcus neoformans isolates is a matter of great concern. The Clinical and Laboratory Standards Institute (CLSI) broth microdilution reference method (BMD) for antifungal susceptibility testing of C. neoformans is tedious and time-consuming. Consequently, there is a greater need for a reproducible in vitro susceptibility testing method for use in clinical microbiology laboratories. By random amplified polymorphic DNA analysis, the 62 Indian clinical isolates were characterized as Cryptococcus neoformans var. grubii. We evaluated the susceptibilities of these isolates for amphotericin B (AMB) and fluconazole (FLC) by two commercial techniques, i.e., Vitek 2 and E-test against the CLSI M27-A3 BMD. The essential agreement (EA) between the Vitek 2 and E-test with the reference procedure for FLC was similar (82.2%). For AMB, EA of 92 and 76% was obtained with E-test and Vitek 2. Excellent categorical agreement (CA) (98.3% and 100% by Vitek 2 and E-test, respectively) was obtained for AMB. The CA for FLC was 81 and 77.4% by Vitek 2 and E-test. We conclude that both E-test and Vitek 2 system have acceptable levels of accuracy for susceptibility testing of both the drugs. Both of them could identify fluconazole-resistant strains. Vitek 2 could be used for testing susceptibility of voriconazole and 5-flucytosine also at the same time.

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References

  1. Banerjee U, Datta K, Majumdar T, Gupta K. Cryptococcosis in India: the awakening of a giant? Med Mycol. 2001;39:51–67.

    PubMed  CAS  Google Scholar 

  2. Shih CC, Chen YC, Chang SC, Luh KT, Hsieh WC. Cryptococcal meningitis in non-HIV-infected patients. Q J Med. 2000;93:245–51.

    Article  CAS  Google Scholar 

  3. Antinori S, Ridolfo A, Fasan M, Magni C, Galimberti L, Milazzo L, Sollima S, Adorni F, Giuliani G, Galli M, Corbellino M, Parravicini C. AIDS-associated cryptococcosis: a comparison of epidemiology, clinical features and outcome in the pre- and post-HAART eras. Experience of a single centre in Italy. 2009. HIV Med. 2009;10:6–11.

    Article  PubMed  CAS  Google Scholar 

  4. Friese G, Discher T, Fussle R, Schmalreck A, Lohmeyer J. Development of azole resistance during fluconazole maintenance therapy for AIDS associated cryptococcal disease. AIDS. 2001;15:2344–5.

    Article  PubMed  CAS  Google Scholar 

  5. Mondon P, Petter R, Amalfitano G, Luzzati R, Concia E, Polacheck I, Kwon-Chung KJ. Heteroresistance to fluconazole and voriconazole in Cryptococcus neoformans. Antimicrob Agents Chemother. 1999;49:1856–61.

    Google Scholar 

  6. Silva PR, Rabelo RA, Terra AP, Teixeira DN. Susceptibility to antifungal agents among Cryptococcus neoformans varieties isolated from patients at a university hospital. Rev Soc Bras Med Trop. 2008;41:158–62.

    PubMed  Google Scholar 

  7. Cuenca-Estrella M, Gomez-Lopez A, Alastruey-Izquierdo A, Bernal-Martinez L, Cuesta I, Buitrago MJ, Rodriguez-Tudela JL. Comparison of the Vitek 2 Antifungal susceptibility system with the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial susceptibility testing (EUCAST) broth microdilution reference methods and with the sensititre yeast one and Etest techniques for in vitro detection of antifungal resistance in yeast isolates. J Clin Microbiol. 2010;48:1782–6.

    Article  PubMed  Google Scholar 

  8. Chryssanthou E, Grönfors C, Khanna N. Comparison of broth macrodilution, broth microdilution and E-test susceptibility tests of Cryptococcus neoformans for fluconazole. Mycoses. 1997;40:423–7.

    Article  PubMed  CAS  Google Scholar 

  9. Bourgeois N, Dehandschoewercker L, Bertout S, et al. Antifungal susceptibility of 205 Candida spp. isolated primarily during invasive Candidiasis and comparison of the Vitek 2 system with the CLSI broth microdilution and Etest methods. J Clin Microbiol. 2010;48:154–61.

    Article  PubMed  CAS  Google Scholar 

  10. González MT, González A, Cantón E, Aller AI, Romero A, Pemán J, Quindós G, Martín-Mazuelos E. Evaluation of the VITEK 2 system to test the susceptibility of Candida spp., Trichosporon asahii and Cryptococcus neoformans to amphotericin B, flucytosine, fluconazole and voriconazole: a comparison with the M27–A3 reference method. Med Mycol. 2010;48:710–9.

    Article  Google Scholar 

  11. Kwon-Chung KJ, Polacheck I, Popkin TJ. Melanin lacking mutants of Cryptococcus neoformans and their virulence for mice. J Bacteriol. 1982;150:1414–21.

    PubMed  CAS  Google Scholar 

  12. Franzot SP, Hamdan JS, Currie BP, Casadevall A. Molecular epidemiology of Cryptococcus neoformans in Brazil and the United States evidence for both local genetic difference and a global clonal population structure. J Clin Microbiol. 1997;35:2243–51.

    PubMed  CAS  Google Scholar 

  13. Meyer W, Mitchell TG, Freedman EZ, Vilgalys R. Hybridization probes for conventional DNA fingerprinting used as single primers in the polymerase chain reaction to distinguish strains of Cryptococcus neoformans. J Clin Microbiol. 1993;31:2274–80.

    PubMed  CAS  Google Scholar 

  14. Clinical and Laboratory Standards Institute. Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved Standard third edition, M27-A3.Wayne, PA; 2008.

  15. Lozano-Chiu M, Paetznick VL, Ghannoum MA, Rex JH. Detection of resistance to amphotericin B among Cryptococcus neoformans clinical isolates: performances of three different media assessed by using E-test and National Committee for Clinical Laboratory Standards M27-A methodologies. J Clin Microbiol. 1998;36:2817–22.

    PubMed  CAS  Google Scholar 

  16. Aller AI, Martín-Mazuelos E, Gutiérrez MJ, Bernal S, Chávez M, Recio FJ. Comparison of the Etest and microdilution method for antifungal susceptibility testing of Cryptococcus neoformans to four antifungal agents. J Antimicrob Chemother. 2000;46:997–1000.

    Article  PubMed  CAS  Google Scholar 

  17. Jain N, Wickes BL, Keller SM, Casadevall A, Jain P, Ragan MA, Banerjee U, Fries BC. Molecular epidemiology of clinical Cryptococcus neoformans strains from India. J Clin Microbiol. 2005;43:5733–42.

    Article  PubMed  CAS  Google Scholar 

  18. Mitchell TG, Perfect JR. Cryptococcosis in the era of AIDS-100 years after the discovery of Cryptococcus neoformans. Clin Microbiol Rev. 1995;8:515–48.

    PubMed  CAS  Google Scholar 

  19. Clark RA, Greer DL, Atkinson W, Valainis GT, Hyslop N. Spectrum of Cryptococcus neoformans infection in 68 patients infected with human immunodeficiency virus. Rev Infect Dis. 1990;12:768–77.

    Article  PubMed  CAS  Google Scholar 

  20. Brandt ME, Pfaller MA, Hajjeh RA, Hajjeh RA, Hamill RJ, Pappas PG, Reingold AL, et al. Trends in antifungal drug susceptibility of Cryptococcus neoformans isolates in the United States: 1992 to 1994 and 1996 to 1998. Antimicrob Agents Chemother. 2001;45:3065–9.

    Article  PubMed  CAS  Google Scholar 

  21. Brandt ME, Pfaller MA, Hajjeh RA, Graviss EA, Rees J, Spitzer ED, Pinner RW, Mayer LW, Cryptococcal Disease Active Surveillance Group. Molecular subtypes and antifungal susceptibilities of serial Cryptococcus neoformans isolates in human immunodeficiency virus associated Cryptococcosis. J Infect Dis. 1996;174:812–20.

    Article  PubMed  CAS  Google Scholar 

  22. Spitzer ED, Spitzer SG, Freundlich LF, Casadevall A. Persistence of initial infection in recurrent Cryptococcus neoformans meningitis. Lancet. 1993;341:595–6.

    Article  PubMed  CAS  Google Scholar 

  23. Pfaller M, Zhang J, Messer S, Tumberland M, Mbidde E, Jessup C, Ghannoum M. Molecular epidemiology and antifungal susceptibility of Cryptococcus neoformans isolates from Ugandan AIDS patients. Diagn Microbiol Infect Dis. 1998;32:191–9.

    Article  PubMed  CAS  Google Scholar 

  24. Franzot SP, Hamdan JS. In vitro susceptibilities of clinical and environmental isolates of Cryptococcus neoformans to five antifungal drugs. Antimicrob Agents Chemother. 1996;40:822–4.

    PubMed  CAS  Google Scholar 

  25. Klepser ME, Pfaller MA. Variation in electrophoretic karyotype and antifungal susceptibility of clinical isolates of Cryptococcus neoformans at a university affiliated teaching hospital from 1987 to 1994. J Clin Microbiol. 1998;36:3653–6.

    PubMed  CAS  Google Scholar 

  26. Capoor MR, Mandal P, Deb M, Aggarwal P, Banerjee U. Current scenario of cryptococcosis and antifungal susceptibility pattern in India: a cause for reappraisal. Mycoses. 2008;51:258–65.

    Article  PubMed  CAS  Google Scholar 

  27. Datta K, Jain N, Sethi S, Rattan A, Casadevall A, Banerjee U. Fluconazole and itraconazole susceptibility of clinical isolates of Cryptococcus neoformans at a tertiary care centre in India: a need for care. J Antimicrob Chemother. 2003;52:683–6.

    Article  PubMed  CAS  Google Scholar 

  28. Colombo AL, Barchiesi F, McGough DA, Rinaldi MG. Comparison of E-test and National Committee for Clinical Laboratory Standards broth macrodilution method for azole antifungal susceptibility testing. J Clin Microbiol. 1995;33:535–40.

    PubMed  CAS  Google Scholar 

  29. Espinel-Ingroff A, Pfaller M, Erwin ME, Jones RN. Interlaboratory evaluation of E-test method for testing antifungal susceptibilities of pathogenic yeasts to five antifungal agents by using Casitone agar and solidified RPMI 1640 medium with 2% glucose. J Clin Microbiol. 1996;34:848–52.

    PubMed  CAS  Google Scholar 

  30. Dias AL, Matsumoto FE, Melhem MS, da Silva EG, Auler ME, de Siqueira AM, Paula CR. Comparative analysis of Etest and broth microdilution method (AFST-EUCAST) for trends in antifungal drug susceptibility testing of Brazilian Cryptococcus neoformans isolates. J Med Microbiol. 2006;55:1693–9.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Microbiology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India

    Aarti Tewari, Bijayani Behera & Immaculata Xess

  2. Department of Lab Medicine, Jpna Trauma Centre, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India

    Purva Mathur

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  1. Aarti Tewari
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  2. Bijayani Behera
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  3. Purva Mathur
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  4. Immaculata Xess
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Correspondence to Immaculata Xess.

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Tewari, A., Behera, B., Mathur, P. et al. Comparative Analysis of the Vitek 2 Antifungal Susceptibility System and E-test with the CLSI M27-A3 Broth Microdilution Method for Susceptibility Testing of Indian Clinical Isolates of Cryptococcus neoformans . Mycopathologia 173, 427–433 (2012). https://doi.org/10.1007/s11046-012-9528-9

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