title: Seeded Amplification of Chronic Wasting Disease Prions in Nasal Brushings and Recto-anal Mucosa-Associated Lymphoid Tissues from Elk by Real-Time Quaking-Induced Conversion.
abstract: Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly 50 years ago in Colorado and Wyoming and has since been detected across North America 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 its spread; this is especially true in cases of relocation/reintroduction or prevalence studies of large or protected herds, where depopulation may be contraindicated. This study evaluated the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay of recto-anal mucosa-associated lymphoid tissue (RAMALT) biopsy specimens and nasal brushings collected antemortem. These findings were compared to results of immunohistochemistry (IHC) analysis of ante- and postmortem samples. RAMALT samples were collected from populations of farmed and free-ranging Rocky Mountain elk (Cervus elaphus nelsoni;n= 323), and nasal brush samples were collected from a subpopulation of these animals (n= 205). We hypothesized that the sensitivity of RT-QuIC would be comparable to that of IHC analysis of RAMALT and would correspond to that of IHC analysis of postmortem tissues. We found RAMALT sensitivity (77.3%) to be highly correlative between RT-QuIC and IHC analysis. Sensitivity was lower when testing nasal brushings (34%), though both RAMALT and nasal brush test sensitivities were dependent on both thePRNPgenotype and disease progression determined by the obex score. These data suggest that RT-QuIC, like IHC analysis, is a relatively sensitive assay for detection of CWD prions in RAMALT biopsy specimens and, with further investigation, has potential for large-scale and rapid automated testing of antemortem samples for CWD.
authors.affiliation: Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, USA nicholas.j.haley@gmail.com.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Haley
authors.affiliation: Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, USA.
authors.forename: Chris
authors.initials: C
authors.lastname: Siepker
authors.affiliation: Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.
authors.forename: Laura L
authors.initials: LL
authors.lastname: Hoon-Hanks
authors.affiliation: Canadian Food Inspection Agency, National and OIE Reference Laboratory for Scrapie and CWD, Ottawa Laboratory Fallowfield, Ottawa, Ontario, Canada.
authors.forename: Gordon
authors.initials: G
authors.lastname: Mitchell
authors.affiliation: U.S. Geological Survey, Pennsylvania Cooperative Fish and Wildlife Research Unit, The Pennsylvania State University, University Park, Pennsylvania, USA.
authors.forename: W David
authors.initials: WD
authors.lastname: Walter
authors.affiliation: TSE/Prion Biochemistry Section, Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana, USA.
authors.forename: Matteo
authors.initials: M
authors.lastname: Manca
authors.affiliation: National Park Service, Biological Resources Division, Fort Collins, Colorado, USA.
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: Monello
authors.affiliation: National Park Service, Biological Resources Division, Fort Collins, Colorado, USA.
authors.forename: Jenny G
authors.initials: JG
authors.lastname: Powers
authors.affiliation: National Park Service, Biological Resources Division, Fort Collins, Colorado, USA.
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Wild
authors.affiliation: Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.
authors.forename: Edward A
authors.initials: EA
authors.lastname: Hoover
authors.affiliation: TSE/Prion Biochemistry Section, Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana, USA.
authors.forename: Byron
authors.initials: B
authors.lastname: Caughey
authors.affiliation: Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, USA.
authors.forename: Jürgen A
authors.initials: JA
authors.lastname: Richt
mesh_terms: D000818:Animals; D001706:Biopsy; D003955:Diagnostic Tests, Routine / Q000379:methods*; D005260:Female; D007413:Intestinal Mucosa / Q000737:chemistry*; D008221:Lymphoid Tissue / Q000737:chemistry*; D008297:Male; D009297:Nasal Mucosa / Q000737:chemistry*; D057089:Pathology, Molecular / Q000379:methods*; D011328:Prions / Q000032:analysis*; D012418:Ruminants; D012680:Sensitivity and Specificity; D013997:Time Factors; D034081:Wasting Disease, Chronic / Q000175:diagnosis*
chemical_list: D011328:Prions