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Journal of Clinical Microbiology, July 2001, p. 2513-2517, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2513-2517.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Standardization of Antifungal Susceptibility Variables for a Semiautomated Methodology

Juan L. Rodríguez-Tudela,* Manuel Cuenca-Estrella, Teresa M. Díaz-Guerra, and Emilia Mellado

Servicio de Micología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, 28220 Majadahonda, Spain

Received 22 January 2001/Returned for modification 2 April 2001/Accepted 13 May 2001

Recently, the methodology that will serve as a basis of the standard for antifungal susceptibility testing of fermentative yeasts of the European Committee on Antibiotic Susceptibility Testing has been described. This procedure employs a spectrophotometric method for both inoculum adjustment and endpoint determination. However, the utilization of a spectrophotometer requires studies for standardization. The present work analyzes the following parameters: (i) accuracy of inoculum preparation, (ii) correlation between optical density and CFU per milliliter, (iii) influence of the wavelength on the endpoint determination, and (iv) influence of the dimethyl sulfoxide concentration on the growth kinetics. The main results can be summarized as follows: (i) inoculum preparation following the methodology recommended by the National Committee for Clinical Laboratory Standards is an exact procedure; (ii) the relationship between optical density and CFU per milliliter is linear (coefficient of determination, r2 = 0.84); (iii) MICs obtained by means of spectrophotometric readings at different wavelengths are identical (for amphotericin B, an intraclass correlation coefficient of 0.98 was obtained; for fluconazole, the intraclass correlation coefficient was 1); and (iv) a 2% concentration of dimethyl sulfoxide produces a significantly slower and lower growth curve of Candida spp. than other concentrations.

* Corresponding author. Mailing address: Servicio de Micología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra Majadahonda-Pozuelo km. 2, 28220 Majadahonda, Spain. Phone: 34 91 5097961. Fax: 34 91 5097966. E-mail: juanl.rodriguez-tudela{at}isciii.es.

Journal of Clinical Microbiology, July 2001, p. 2513-2517, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2513-2517.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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