Detection of CWD in cervids by RT-QuIC assay of third eyelids
RESEARCH ARTICLE
Detection of CWD in cervids by RT-QuIC assay of third eyelids
Sarah K. Cooper1 , Clare E. Hoover1¤ , Davin M. HendersonID1 , Nicholas J. Haley2 , Candace K. Mathiason1 , Edward A. Hoover1 * 1 Prion Research Center, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, United States of America, 2 Department of Microbiology and Immunology, Midwestern University, Glendale, Arizona, United States of America ¤ Current address: AstraZeneca, Waltham, Massachusetts, United States of America * edward.hoover@colostate.edu
Abstract
The diagnosis of chronic wasting disease (CWD) relies on demonstration of the diseaseassociated misfolded CWD prion protein (PrPCWD) in brain or retropharyngeal lymph node tissue by immunodetection methods, e.g. ELISA and immunohistochemistry (IHC). The success of these methods relies on a quality sample of tissues, which requires both anatomical knowledge and considerable dissection to collect. As the prevalence of CWD continues to increase globally, the development of fast and cost-effective methods to detect the disease is vital to facilitate CWD detection and surveillance. To address these issues, we have evaluated third eyelids from CWD-infected deer and elk using real-time quaking induced conversion (RT-QuIC). We identified prion seeding activity in third eyelids in 24 of 25 (96%) CWDinfected white-tailed deer (Odocoileus virginianus). We detected RT-QuIC positivity in the third eyelid as early as 1 month after experimental CWD exposure. In addition, we identified prion seeding activity in third eyelids of 18 of 25 (72%) naturally exposed asymptomatic CWD-positive rocky mountain elk (Cervus canadensis nelson). We compared CWD detection by RT-QuIC and IHC in third eyelid, retropharyngeal lymph node, and brain in 10 deer in early symptomatic stage of disease. IHC detected PrPCWD deposition in third eyelid lymphoid follicles in 5 of 10 deer (50%) whereas third eyelids of all 10 animals were positive by RT-QuIC. This difference reflected in part a lower requirement for lymphoid follicle presence for seeding activity detection by RT-QuIC. In conclusion, RT-QuIC analysis of the third eyelid, an easily accessed tissue, has potential to advance CWD detection and testing compliance.
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In conclusion, we demonstrate that RT-QuIC performed on third eyelid tissue can be used to detect CWD in deer and elk, including those in pre-symptomatic stages of infection. As third eyelid is an easily accessible tissue, it has potential to aid in surveillance and screening programs
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posted by Terry S. Singeltary Sr. at
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