Researchers from Indiana University developed a rapid multiplex real-time PCR assay capable of detecting Histoplasma, Blastomyces, and Coccidioides directly from clinical specimens. Presented at AMP 2025, the test bypasses slow culture and often inconclusive serology by targeting ITS1, BAD1, and A2/PRA gene regions unique to each fungus. Running on the DiaSorin LIAISON MDX platform with a simplified lysis method, the assay showed 100% agreement with standard-of-care testing across 32 specimens and demonstrated complete analytical specificity. Early data suggest it meets CAP requirements and could sharply improve turnaround times for endemic fungal diagnoses, particularly in immunocompromised patients.