Sunday, February 14, 2016

Antemortem detection of chronic wasting disease prions in nasal brush collections and rectal biopsies from white-tailed deer by real time quaking-induced conversion

Antemortem detection of chronic wasting disease prions in nasal brush collections and rectal biopsies from white-tailed deer by real time quaking-induced conversion


Nicholas J. Haleya#, Chris Siepkera, W. David Walterb, Bruce V. Thomsenc, Justin J. Greenleed, Aaron D. Lehmkuhlc and Jürgen A. Richta


+ Author Affiliations Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, USAa U.S. Geological Survey, Pennsylvania Cooperative Fish and Wildlife Research Unit, Pennsylvania State University, University Park, Pennsylvania, USAb 3USDA, APHIS, VS, STAS, National Veterinary Service Laboratories, Ames, Iowa, USAc Virus and Prion Research Unit, National Animal Disease Center, ARS, USDA, Ames, Iowa, USAd




Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly fifty years ago in Colorado and Wyoming, and has since spread to cervids in 23 states, two Canadian provinces, and the Republic of Korea. The expansion of this disease makes the development of sensitive diagnostic assays and antemortem sampling techniques crucial for the mitigation of spread; this is especially true in cases of relocation/reintroduction of farmed or free-ranging deer and elk, or surveillance studies in private or protected herds where depopulation is contraindicated. This study sought to evaluate the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay using recto-anal mucosa associated lymphoid tissue (RAMALT) biopsies and nasal brush samples collected antemortem from farmed white-tailed deer (n=409). Antemortem findings were then compared to results from ante- and postmortem samples (RAMALT, brainstem and medial retropharyngeal lymph nodes) evaluated using the current gold standard in vitro assay, immunohistochemistry (IHC). We hypothesized the sensitivity of RT-QuIC would be comparable to IHC in antemortem tissues, and would correlate with both genotype and stage of clinical disease. Our results showed that RAMALT testing by RT-QuIC had the highest sensitivity (69.8%) when compared to postmortem testing, with a specificity of >93.9%. These data suggest that RT-QuIC, like IHC, is an effective assay for detection of PrPCWD in rectal biopsies and other antemortem samples, and with further research to identify more sensitive tissues, bodily fluids, or experimental conditions, has potential for large scale and rapid automated testing for CWD diagnosis.




↵#Address correspondence to Nicholas J. Haley, Copyright © 2016, American Society for Microbiology. All Rights Reserved.



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