Identification of Histoplasma capsulatum, Blastomyces dermatitidis, and Coccidioides species by repetitive-sequence-based PCR.
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 > or =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 > or =90% similarity was used for identification. One H. capsulatum isolate could not be identified; no isolates were misidentified. Using > or =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.
Pounder
June I
JI
Associated Regional and University Pathologists, Inc., ARUP Laboratories, Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT 84108. june.pounder@aruplab.com
Hansen
Dewey
D
Woods
Gail L
GL
D001758:Blastomyces / Q000145:classification* / Q000166:cytology / Q000235:genetics / Q000254:growth & development; D016000:Cluster Analysis; D003045:Coccidioides / Q000145:classification* / Q000166:cytology / Q000235:genetics / Q000254:growth & development; D016172:DNA Fingerprinting / Q000379:methods*; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D006658:Histoplasma / Q000145:classification* / Q000166:cytology / Q000235:genetics / Q000254:growth & development; D006801:Humans; D008853:Microscopy; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012680:Sensitivity and Specificity
D004271:DNA, Fungal