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Journal of Clinical Microbiology, March 2006, p. 693-699, Vol. 44, No. 3
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.3.693-699.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of Medically Important Yeast Species by Sequence Analysis of the Internal Transcribed Spacer Regions

Shiang Ning Leaw,1 Hsien Chang Chang,1 Hsiao Fang Sun,2 Richard Barton,3 Jean-Philippe Bouchara,4 and Tsung Chain Chang5*

Institute of Biomedical Engineering,1 Institute of Molecular Medicine,2 Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, Taiwan, Republic of China,5 School of Biochemistry and Microbiology, University of Leeds, Leeds, United Kingdom,3 Host-Parasite-Interaction Study Group (UPRES-EA 3142), Laboratory of Parasitology and Mycology, University Hospital, Angers, France4

Received 18 July 2005/ Returned for modification 27 October 2005/ Accepted 13 December 2005

Infections caused by yeasts have increased in previous decades due primarily to the increasing population of immunocompromised patients. In addition, infections caused by less common species such as Pichia, Rhodotorula, Trichosporon, and Saccharomyces spp. have been widely reported. This study extensively evaluated the feasibility of sequence analysis of the rRNA gene internal transcribed spacer (ITS) regions for the identification of yeasts of clinical relevance. Both the ITS1 and ITS2 regions of 373 strains (86 species), including 299 reference strains and 74 clinical isolates, were amplified by PCR and sequenced. The sequences were compared to reference data available at the GenBank database by using BLAST (basic local alignment search tool) to determine if species identification was possible by ITS sequencing. Since the GenBank database currently lacks ITS sequence entries for some yeasts, the ITS sequences of type (or reference) strains of 15 species were submitted to GenBank to facilitate identification of these species. Strains producing discrepant identifications between the conventional methods and ITS sequence analysis were further analyzed by sequencing of the D1-D2 domain of the large-subunit rRNA gene for species clarification. The rates of correct identification by ITS1 and ITS2 sequence analysis were 96.8% (361/373) and 99.7% (372/373), respectively. Of the 373 strains tested, only 1 strain (Rhodotorula glutinis BCRC 20576) could not be identified by ITS2 sequence analysis. In conclusion, identification of medically important yeasts by ITS sequencing, especially using the ITS2 region, is reliable and can be used as an accurate alternative to conventional identification methods.

* Corresponding author. Mailing address: Department of Medical Laboratory Science and Biotechnology, School of Medicine, National Cheng Kung University, 1 University Road, Tainan 701, Taiwan, Republic of China. Phone: 886-6-2353535, ext. 5790. Fax: 886-6-2363956. E-mail: tsungcha{at}mail.ncku.edu.tw.

Journal of Clinical Microbiology, March 2006, p. 693-699, Vol. 44, No. 3
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.3.693-699.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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