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Journal of Clinical Microbiology, August 2006, p. 2977-2982, Vol. 44, No. 8
0095-1137/06/$08.00+0     doi:10.1128/JCM.00687-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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

Associated Regional and University Pathologists, Inc., Institute for Clinical and Experimental Pathology, Salt Lake City, Utah,¹ ARUP Microbiology Laboratory, Salt Lake City, Utah,² Department of Pathology and Laboratory Services, University of Arkansas for Medical Sciences, Little Rock, Arkansas³

Received 31 March 2006/ Returned for modification 15 May 2006/ Accepted 31 May 2006

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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