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Journal of Clinical Microbiology, November 2003, p. 5099-5102, Vol. 41, No. 11
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.11.5099-5102.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Experience with the MicroSeq D2 Large-Subunit Ribosomal DNA Sequencing Kit for Identification of Commonly Encountered, Clinically Important Yeast Species

Leslie Hall, Sherri Wohlfiel, and Glenn D. Roberts*

Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota 55905

Received 8 April 2003/ Returned for modification 24 June 2003/ Accepted 4 August 2003

Experience with a MicroSeq D2 large-subunit (LSU) ribosomal DNA (rDNA) sequencing kit for identification of yeast species commonly encountered in the mycology laboratory at Mayo Clinic is described here. A total of 131 isolates of yeasts recovered from clinical specimens were included in the study. Phenotypic methods used for initial identification included germ tube formation, urease production, microscopic morphological features on cornmeal agar, and an API 20C AUX system; all isolates were sequenced using a MicroSeq D2 LSU rDNA sequencing kit. Nucleic acid sequencing identified 93.9% of the isolates to the correct genus and species. A total of 100 of the isolates (representing 19 species of Candida) were sequenced, and 98% gave results concordant with identifications made by the API 20C AUX system; distance scores ranged from 0 to 1.88%, with an average value of 0.23%. Candida dubliniensis was not included in the MicroSeq database and was identified as Candida albicans. A total of 32 isolates representing 9 other genera (including Cryptococcus, Filobasidium, Kloeckera, Malassezia, Pichia, Sporidiobolus, Rhodotorula, Zygosaccharomyces, and Trichosporon) were included, and 81.3% showed concordant results when phenotypic and sequencing results were compared. Most discrepancies were attributed to the lack of inclusion of the species in the MicroSeq or API 20C AUX database. The MicroSeq D2 LSU rDNA sequencing kit appears to be accurate and useful for the identification of yeasts that might be seen in a clinical laboratory.

* Corresponding author. Mailing address: Division of Clinical Microbiology, Mayo Clinic, 200 First St. SW, Rochester, MN 55905. Phone: (507) 284-3704. Fax: (507) 284-9859. E-mail: roberts.glenn{at}mayo.edu.

Journal of Clinical Microbiology, November 2003, p. 5099-5102, Vol. 41, No. 11
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.11.5099-5102.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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