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Identification of Medically Important Candida and Non-Candida Yeast Species by an Oligonucleotide Array

Institute of Biomedical Engineering,¹ Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, Taiwan, Republic of China,⁴ Institute of Molecular and Cellular Biology, University of Leeds, United Kingdom,² Host-Parasite Interaction Study Group, UPRES-EA 3142, Laboratory of Parasitology and Mycology, Angers University Hospital, Angers, France³

Received 12 March 2007/ Returned for modification 30 March 2007/ Accepted 7 May 2007

The incidence of yeast infections has increased in the recent decades, with Candida albicans still being the most common cause of infections. However, infections caused by less common yeasts have been widely reported in recent years. Based on the internal transcribed spacer 1 (ITS 1) and ITS 2 sequences of the rRNA genes, an oligonucleotide array was developed to identify 77 species of clinically relevant yeasts belonging to 16 genera. The ITS regions were amplified by PCR with a pair of fungus-specific primers, followed by hybridization of the digoxigenin-labeled PCR product to a panel of oligonucleotide probes immobilized on a nylon membrane for species identification. A collection of 452 yeast strains (419 target and 33 nontarget strains) was tested, and a sensitivity of 100% and a specificity of 97% were obtained by the array. The detection limit of the array was 10 pg of yeast genomic DNA per assay. In conclusion, yeast identification by the present method is highly reliable and can be used as an alternative to the conventional identification methods. The whole procedure can be finished within 24 h, starting from isolated colonies.

Published ahead of print on 16 May 2007.

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