Digging into soil: Effects of soil texture on RT-QuIC performance for environmental prion surveillance.

Chronic wasting disease (CWD) is a fatal neurodegenerative disease caused by infectious prions affecting wild and captive cervids. Transmission occurs directly between hosts or indirectly through contact with prion-contaminated environments. Soils, particularly those rich in clay, are hypothesized to enhance prion stability, retention, and bioavailability. Accurate detection of prions is therefore important for understanding environmental transmission risks. Real-time quaking-induced conversion (RT-QuIC) is a sensitive assay used to detect PrP^CWD in tissue, excreta, and environmental materials. However, RT-QuIC performance across soil textures has not been evaluated. This study assessed RT-QuIC sensitivity and specificity using laboratory-prepared soils spiked with CWD-positive brain homogenate or water controls under a standardized extraction method. Conditional on the extraction method used, results suggest that RT-QuIC performance depends on soil texture, and thus, an optimal time-to-threshold (TTT) cutoff required to balance sensitivity and specificity will also vary with soil texture. RT-QuIC exhibited higher sensitivity and moderate specificity in soils with low clay (<20%) and moderate to high silt content, whereas high-clay soils (>20%) with low to moderate silt content (2%–60%) reduced both sensitivity and specificity, and required shorter TTT cutoffs. These findings highlight the importance of accounting for soil texture in environmental CWD surveillance.