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Journal of Clinical Microbiology, November 2001, p. 4076-4081, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.4076-4081.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Analysis of Microsatellite Markers of Candida albicans Used for Rapid Typing

Françoise Botterel,1,2 Christophe Desterke,1 Catherine Costa,1 and Stéphane Bretagne1,*

Laboratoire de Parasitologie-Mycologie, Hôpital H. Mondor (AP-HP) et Université Paris XII, 94010 Créteil,1 and Laboratoire de Parasitologie-Mycologie, Hôpital du Kremlin-Bicêtre (AP-HP), Kremlin-Bicêtre,2 France

Received 22 June 2001/Returned for modification 26 July 2001/Accepted 12 August 2001

To obtain a rapid genotyping method of Candida albicans, three polymorphic microsatellite markers were investigated by multiplex PCR. The three loci, called CDC3, EF3, and HIS3, were chosen because they are on different chromosomes so as to improve the chances of finding polymorphisms. One set of primers was designed for each locus, and one primer of each set was dye-labeled to read PCR signals by using an automatic sequencer. Amplifications were performed directly from the colonies harvested on the agar plate without a sophisticated DNA extraction step. At total of 27 reference strains and 73 clinical independent isolates were tested. The numbers of allelic associations were 10, 22, and 25 for the loci CDC3, EF3, and HIS3, respectively. The combined discriminatory power of the three microsatellites markers was 0.97. The markers were stable after 25 subcultures, and the amplifications were specific for C. albicans. An initial study of 17 clinical isolate pairs, including blood culture and peripheral sites, showed a similar genotype for 15 of them, confirming that candidemia usually originates from the colonizing isolate. Therefore, microsatellite marker analysis with multiplex PCR and automated procedures has a high throughput and should be suitable for large epidemiologic studies of C. albicans.

* Corresponding author. Mailing address: Laboratoire de Parasitologie-Mycologie, Hôpital Henri Mondor, 51 avenue du Maréchal DeLattre de Tassigny, 94010 Créteil Cedex, France. Phone: (33) 01-49-81-36-41. Fax: (33) 01-49-81-36-01. E-mail: bretagne{at}univ-paris12.fr.

Journal of Clinical Microbiology, November 2001, p. 4076-4081, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.4076-4081.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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