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Journal of Clinical Microbiology, April 2000, p. 1599-1608, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Evaluation of Phenotypic Markers for Selection and Identification of Candida dubliniensis

Kathrin Tintelnot,1,* Gerhard Haase,2 Michael Seibold,1 Frank Bergmann,3 Maren Staemmler,1 Tatjana Franz,1 and Dieter Naumann1

Robert Koch-Institute1 and Medizinische Klinik (Schwerpunkt Infektiologie) der Charité, Campus Virchow-Klinikum, Humboldt University,3 Berlin, and Institute of Medical Microbiology, University Hospital RWTH Aachen, Aachen,2 Germany

Received 16 June 1999/Returned for modification 2 October 1999/Accepted 27 December 1999

Candida dubliniensis is often associated with C. albicans in cultures. Easy-to-perform selective isolation procedures for these closely related species do not exist. Therefore, we evaluated previously described discriminatory phenotypic markers for C. dubliniensis. A total of 150 oral rinses from human immunodeficiency virus (HIV)-infected patients were cultured on CHROMagar Candida. Dark green colonies described as being indicative of C. dubliniensis and other green colonies, 170 in total, were isolated. Chlamydospore formation, intracellular beta -D-glucosidase activity, ability to grow at 42°C, carbohydrate assimilation pattern obtained by the API ID 32C, and Fourier transform infrared (FT-IR) spectroscopy were used for phenotypic characterization. Sequencing of the 5' end of the nuclear large-subunit (26S) ribosomal DNA gene was used for definitive species identification for C. dubliniensis. C. dubliniensis was found in 34% of yeast-colonized HIV-infected patients. The color of the colonies on CHROMagar Candida proved to be insufficient for selecting C. dubliniensis, since only 30 of 53 proven C. dubliniensis isolates showed a dark green color in primary cultures. The described typical chlamydospore formation can give only some indication of C. dubliniensis. The assimilation pattern proved to be insufficient to discriminate C. dubliniensis from C. albicans. All C. dubliniensis strains showed no or highly restricted growth at 42°C and a lack of beta -D-glucosidase activity. Unfortunately, atypical C. albicans strains can also exhibit these phenotypic traits. FT-IR spectroscopy combined with hierarchical clustering proved to be as reliable as genotyping for discriminating the two species.

* Corresponding author. Mailing address: Robert Koch-Institute, Nordufer 20, D-13353 Berlin, Germany. Phone: 49 18 88754 2208. Fax: 49 18 88754 2614. E-mail: tintelnotk{at}rki.de.

Journal of Clinical Microbiology, April 2000, p. 1599-1608, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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