Scrapie versus Chronic Wasting Disease in White-Tailed Deer.

White-tailed deer are susceptible to scrapie (WTD scrapie) after oronasal inoculation with the classical scrapie agent from sheep. Deer affected by WTD scrapie are difficult to differentiate from deer infected with chronic wasting disease (CWD). To assess the transmissibility of the WTD scrapie agent and tissue phenotypes when further passaged in white-tailed deer, we oronasally inoculated wild-type white-tailed deer with WTD scrapie agent. We found that WTD scrapie and CWD agents were generally similar, although some differences were noted. The greatest differences were seen in bioassays of cervidized mice that exhibited significantly longer survival periods when inoculated with WTD scrapie agent than those inoculated with CWD agent. Our findings establish that white-tailed deer are susceptible to WTD scrapie and that the presence of WTD scrapie agent in the lymphoreticular system suggests the handling of suspected cases should be consistent with current CWD guidelines because environmental shedding may occur.

Large-scale validation of skin prion seeding activity as a biomarker for diagnosis of prion diseases.

Definitive diagnosis of sporadic Creutzfeldt-Jakob disease (sCJD) relies on the examination of brain tissues for the pathological prion protein (PrPSc). Our previous study revealed that PrPSc-seeding activity (PrPSc-SA) is detectable in skin of sCJD patients by an ultrasensitive PrPSc seed amplification assay (PrPSc-SAA) known as real-time quaking-induced conversion (RT-QuIC). A total of 875 skin samples were collected from 2 cohorts (1 and 2) at autopsy from 2-3 body areas of 339 cases with neuropathologically confirmed prion diseases and non-sCJD controls. The skin samples were analyzed for PrPSc-SA by RT-QuIC assay. The results were compared with demographic information, clinical manifestations, cerebrospinal fluid (CSF) PrPSc-SA, other laboratory tests, subtypes of prion diseases defined by the methionine (M) or valine (V) polymorphism at residue 129 of PrP, PrPSc types (#1 or #2), and gene mutations in deceased patients. RT-QuIC assays of the cohort #1 by two independent laboratories gave 87.3% or 91.3% sensitivity and 94.7% or 100% specificity, respectively. The cohort #2 showed sensitivity of 89.4% and specificity of 95.5%. RT-QuIC of CSF available from 212 cases gave 89.7% sensitivity and 94.1% specificity. The sensitivity of skin RT-QuIC was subtype dependent, being highest in sCJDVV1-2 subtype, followed by VV2, MV1-2, MV1, MV2, MM1, MM1-2, MM2, and VV1. The skin area next to the ear gave highest sensitivity, followed by lower back and apex of the head. Although no difference in brain PrPSc-SA was detected between the cases with false negative and true positive skin RT-QuIC results, the disease duration was significantly longer with the false negatives [12.0 ± 13.3 (months, SD) vs. 6.5 ± 6.4, p < 0.001]. Our study validates skin PrPSc-SA as a biomarker for the detection of prion diseases, which is influenced by the PrPSc types, PRNP 129 polymorphisms, dermatome sampled, and disease duration.

Transmission of classical scrapie using lymph node inoculum.

Scrapie is a fatal, transmissible neurodegenerative disease that affects sheep and goats. Replication of PrPSc in the lymphoid tissue allows for the scrapie agent to be shed into the environment. Brain and retropharyngeal lymph node (RPLN) from a sheep inoculated with the classical scrapie agent was used to compare infectivity of these tissues. Nine Cheviot sheep were used in this study, randomly assigned into two groups based on inocula. Group one (n = 4) received 1 mL of 10% brain homogenate and consisted of all VRQ/VRQ PRNP genotypes. Group two (n = 5) had three sheep receive 1 mL of a 10% RPLN homogenate (13-7), and two sheep receive 0.5 mL of a 10% RPLN homogenate (13-7) because of availability. Sheep in group two were also VRQ/VRQ genotyped. Brain and lymph tissues were tested by histopathology, immunohistochemistry, western blot, enzyme immunoassay, and conformational stability for PrPSc accumulation. Both groups displayed clinical signs of ataxia, moribund, head tremors, circling, and lethargy prior to euthanizing at an average of 16.2 mpi (months post inoculation) (group one) or 19.56 mpi (group two). Additionally, brainstem tissue from both groups displayed the same apparent molecular mass by western blot examination. Spongiform lesion profiling and PrPSc accumulation in brain and lymph tissues were similar in both groups. Conformational stability results displayed no significant difference in obex or RPLN tissue. Overall, these data suggest lymph nodes containing the classical scrapie agent are infectious to sheep, aiding in the understanding of sheep scrapie transmission.