title: Experience with the MicroSeq D2 large-subunit ribosomal DNA sequencing kit for identification of filamentous fungi encountered in the clinical laboratory.
abstract: 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.
authors.affiliation: Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota 55905, USA.
authors.forename: Leslie
authors.initials: L
authors.lastname: Hall
authors.forename: Sherri
authors.initials: S
authors.lastname: Wohlfiel
authors.forename: Glenn D
authors.initials: GD
authors.lastname: Roberts
mesh_terms: D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D009181:Mycoses / Q000145:classification / Q000175:diagnosis*; D011933:Reagent Kits, Diagnostic
chemical_list: D004271:DNA, Fungal; D004275:DNA, Ribosomal; D011933:Reagent Kits, Diagnostic