Evaluating the Diagnostic Efficacy of Using Pooled Samples for Chronic Wasting Disease Testing and Surveillance

Disease monitoring informs the opportunities for intervention by natural resource agencies tasked with managing chronic wasting disease (CWD) in wild cervids. However, allocating funds toward testing can reduce those available for education, outreach, and disease reduction. Implementation of more efficient testing strategies can help meet both an expanding need by resource managers and a burgeoning demand from the hunting public in North America. Here, we evaluated the efficacy of pooled testing using the enzyme-linked immunosorbent assay (ELISA), the current screening test used by veterinary diagnostic laboratories in the United States, and real-time quaking-induced conversion (RT-QuIC), an amplification assay that is being evaluated by the U.S. Department of Agriculture but is not yet approved or commercially available. The samples used in this study consisted of medial retropharyngeal lymph nodes (RPLNs) routinely collected by the Iowa Department of Natural Resources during the 2019-2020 surveillance season. The test pools contained tissue from one positive deer diluted in tissue from an increasing number of undetected deer, with each individual contributing an equal tissue volume. ELISA remained positive with pooling thresholds of 1:1, 1:2, 1:4, and 1:9 at a standard volume of tissue homogenate, whereas RT-QuIC remained positive with pooling thresholds of 1:1, 1:2, 1:4, 1:9, 1:19, and 1:49 at a 0.02% tissue dilution. Our results suggest that pooled testing can reduce diagnostic costs multi-fold, and RT-QuIC can be a viable screening test compatible with current field collection standards.