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Journal of Clinical Microbiology, February 2004, p. 622-626, Vol. 42, No. 2
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.2.622-626.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Experience with the MicroSeq D2 Large-Subunit Ribosomal DNA Sequencing Kit for Identification of Filamentous Fungi Encountered in the Clinical Laboratory

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

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

Received 19 August 2003/ Returned for modification 27 October 2003/ Accepted 13 November 2003

Described herein is our experience with the MicroSeq D2 large-subunit rDNA sequencing kit for the identification of filamentous fungi encountered in the mycology laboratory at the Mayo Clinic. A total of 234 filamentous fungi recovered from clinical specimens were used in the evaluation. All were identified by using phenotypic characteristics as observed macroscopically and microscopically on any medium or a combination of media, which included Sabouraud's dextrose, inhibitory mold, cornmeal, Czapek-Dox, potato dextrose, and V8 juice agars; all isolates were sequenced using the MicroSeq D2 large-subunit rDNA sequencing kit. Of the of 234 isolates, 158 were correctly identified to the appropriate genus or genus and species by using nucleic acid sequencing. Sequences for 70 (29.9%) of the isolates (27 genera) were not included in the MicroSeq library. Of the 80 dematiaceous and 154 hyaline fungi sequenced, 65 and 51.2%, respectively, gave results concordant with those determined by phenotypic identification. Nucleic acid sequencing using the MicroSeq D2 large-subunit rDNA sequencing kit offers promise of being an accurate identification system; however, the associated library needs to include more of the clinically important genera and species.

* Corresponding author. Mailing address: Division of Clinical Microbiology, Mayo Clinic, 200 First St. SW, Rochester, MN 55905. Phone: (507) 284-3704. Fax: (507) 284-4272. E-mail: roberts.glenn{at}mayo.edu.

Journal of Clinical Microbiology, February 2004, p. 622-626, Vol. 42, No. 2
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.2.622-626.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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