New Molecular Test Simultaneously Detects Three Major Fungal Infections

Serious fungal infections associated with soil exposure remain difficult to diagnose promptly, especially in regions where Histoplasma, Blastomyces, and Coccidioides are endemic. Many patients present with symptoms resembling routine respiratory illnesses, and current diagnostic tools rely on culturing, which can take weeks, or antibody tests that often fail in early disease. These delays can put both healthy and immunocompromised individuals at risk of worsening illness. A new molecular test now offers a faster, more accurate way to detect these infections simultaneously.

The multiplex real-time PCR assay, developed by researchers at Indiana University Health (Indianapolis, IN, USA)and the Indiana University School of Medicine (Indianapolis, IN, USA), can detect three major pathogenic fungi from a single clinical specimen. Target sequences included ITS1 for Histoplasma, BAD1 for Blastomyces, and the A2/PRA gene for Coccidioides.

Image: The multiplex molecular assay rapidly identifies three major fungal pathogens causing serious respiratory disease (Photo courtesy of Adobe Stock)

The assay is designed to bypass traditional culture requirements, avoiding delays caused by the fungi’s slow growth and morphological switching. Its workflow also eliminates the need for labor-intensive DNA-extraction steps, reducing hands-on time and minimizing risk when handling highly infectious organisms. The platform is compatible with systems already present in many clinical diagnostic laboratories.

The work, presented at the AMP 2025 Annual Meeting & Expo, compared PCR performance against standard culture-based and antigen-based methods. Testing demonstrated 100% sensitivity across all clinical and reference samples evaluated, with the assay correctly identifying every positive specimen. Specificity also reached 100%, with no false signals from other fungi or contaminants.

The simplified workflow further accelerated processing time by eliminating extraction steps while maintaining analytical precision. These findings suggest the assay can deliver accurate identification long before culture results become available. Overall, the validation highlights its reliability as a front-line molecular diagnostic tool for endemic mycoses.

Improved early detection could enable clinicians to begin antifungal therapy sooner, reduce misdiagnosis, and prevent severe disease progression in high-risk patients. The ability to test for Histoplasma, Blastomyces, and Coccidioides in a single run could streamline workflows in laboratories across endemic regions. Because the platform aligns with equipment already widely used, adoption could be rapid and cost-efficient.

This approach may also serve as a definitive species-level identification method after culture growth when needed. Ongoing clinical validation aims to confirm performance in broader patient populations and across specimen types. Further work will determine the assay’s role in future diagnostic guidelines for fungal infections.

“Our assay has the potential to significantly improve turnaround time and diagnostic confidence for infections that have historically been difficult to detect quickly,” said Dr. Kenneth Gavina, PhD, project lead at Indiana University School of Medicine. “While additional clinical validation is underway, this assay shows strong potential to fill a major gap in fungal diagnostics.”

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