Identification of Histoplasma capsulatum , Blastomyces dermatitidis , and Coccidioides Species by Repetitive-Sequence-Based PCR

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Abstract
The performance of repetitive-sequence-based PCR (rep-PCR) using the DiversiLab system for identification of Coccidioides species, Blastomyces dermatitidis , and Histoplasma capsulatum was assessed by comparing data obtained to colony morphology and microscopic characteristics and to nucleic acid probe results. DNA from cultures of 23 Coccidioides , 24 B. dermatitidis , 24 H. capsulatum , 3 Arthrographis , and 2 Malbranchea isolates was extracted using a microbial DNA isolation kit as recommended by Bacterial Barcodes, Inc. Rep-PCR and probe results agreed for 97.2% of the dimorphic fungi when ≥85% similarity was used as the criterion for identification. Two H. capsulatum isolates were not identified, but no isolates were misidentified. From 43 of those cultures (15 Coccidioides , 14 B. dermatitidis , 14 H. capsulatum , 3 Arthrographis , and 2 Malbranchea ), DNA also was extracted using an IDI lysis kit, a simpler method. Rep-PCR and probe results agreed for 97.7% of the dimorphic fungi when a criterion of ≥90% similarity was used for identification. One H. capsulatum isolate could not be identified; no isolates were misidentified. Using ≥85% similarity for identification resulted in one misidentification. These data suggest that the DiversiLab system can be used to identify Coccidioides and B. dermatitidis and, possibly, H. capsulatum isolates.

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