Multiplex PCR detects Histoplasma, Blastomyces, and Coccidioides in a single run.

Designed using primers for ITS1, BAD1, and A2/PRA—regions unique to each fungus.

Bypasses culture (weeks) and serology, which may be delayed or inconclusive.

Achieved 100% agreement with standard testing in 32 clinical isolates.

Demonstrated 100% analytical specificity against nondimorphic fungi and controls.

Uses simplified lysis, eliminating full extraction and reducing biosafety risk.

Early validation suggests it could meet CAP requirements for fungal identification.