Wednesday, September 18, 2013
Rapid Antemortem Detection of CWD Prions in Deer Saliva
Davin M. Henderson equal contributor, equal contributor Contributed equally to this work with: Davin M. Henderson, Matteo Manca
Affiliation: Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University (CSU), Fort Collins, Colorado, United States of America
Chronic wasting disease (CWD) is an efficiently transmitted prion disease of cervids, now identified in 22 United States, 2 Canadian provinces and Korea. One hallmark of CWD is the shedding of infectious prions in saliva, as demonstrated by bioassay in deer. It is also clear that the concentration of prions in saliva, blood, urine and feces is much lower than in the nervous system or lymphoid tissues. Rapid in vitro detection of CWD (and other) prions in body fluids and excreta has been problematic due to the sensitivity limits of direct assays (western blotting, ELISA) and the presence of inhibitors in these complex biological materials that hamper detection. Here we use real-time quaking induced conversion (RT-QuIC) to demonstrate CWD prions in both diluted and prion-enriched saliva samples from asymptomatic and symptomatic white-tailed deer. CWD prions were detected in 14 of 24 (58.3%) diluted saliva samples from CWD-exposed white-tailed deer, including 9 of 14 asymptomatic animals (64.2%). In addition, a phosphotungstic acid enrichment enhanced the RT-QuIC assay sensitivity, enabling detection in 19 of 24 (79.1%) of the above saliva samples. Bioassay in Tg[CerPrP] mice confirmed the presence of infectious prions in 2 of 2 RT-QuIC-positive saliva samples so examined. The modified RT-QuIC analysis described represents a non-invasive, rapid ante-mortem detection of prions in complex biologic fluids, excreta, or environmental samples as well as a tool for exploring prion trafficking, peripheralization, and dissemination.
Citation: Henderson DM, Manca M, Haley NJ, Denkers ND, Nalls AV, et al. (2013) Rapid Antemortem Detection of CWD Prions in Deer Saliva. PLoS ONE 8(9): e74377. doi:10.1371/journal.pone.0074377
Editor: Ilia V Baskakov, University of Maryland School of Medicine, United States of America
Received: June 11, 2013; Accepted: July 31, 2013; Published: September 11, 2013
Copyright: © 2013 Henderson et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: Supported by NIH NINDS grant R01 NS-061902 and by grant D12ZO-045 from the Morris Animal Foundation. This work was also supported in part by the Intramural Research Program of the NIAID. We gratefully acknowledge the Sardinia Regional Government for the financial support of Matteo Manca through his Ph.D. scholarship (P.O.R. Sardegna F.S.E. Operational Programme of the Autonomous Region of Sardinia, European Social Fund 2007-2013). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Tuesday, September 17, 2013
USAHA 116TH ANNUAL MEETING October 18 – 24, 2012 CWD, Scrapie, BSE, TSE prion (September 17, 2013)