Tonsil biopsy to detect chronic wasting disease in white-tailed deer (Odocoileus virginianus) by immunohistochemistry

Authors: Schneider, David; Lehmkuhl, Aaron; SPRAKER, TERRY - Colorado State University; DITTMAR, ROBERT - Texas Parks And Wildlife; LOCKWOOD, MITCH - Texas Parks And Wildlife; ROLLO, SUSAN - Texas Animal Health Commission; NICHOLS, TRACY - Animal And Plant Health Inspection Service (APHIS)

Submitted to: PLOS ONE
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/13/2023
Publication Date: 3/30/2023
Citation: Schneider, D.A., Lehmkuhl, A.D., Spraker, T.R., Dittmar, R.O., Lockwood, M.A., Rollo, S., Nichols, T.A. 2023. Tonsil biopsy to detect chronic wasting disease in white-tailed deer (Odocoileus virginianus) by immunohistochemistry. PLOS ONE. 18(3). Article e0282356. https://doi.org/10.1371/journal.pone.0282356.
DOI: https://doi.org/10.1371/journal.pone.0282356

Interpretive Summary: Chronic wasting disease (CWD) continues to spread in wild and farmed cervid populations. Live-animal testing of farmed cervids is of considerable interest to producers and regulatory agencies as an early detection tool. Choices for live animal samples are limited but include biopsy of the tonsil. The diagnostic sensitivity of CWD immunohistochemistry for tonsil biopsies from white-tailed deer was determined in this study. The overall sensitivity was 92% for deer in a late infection stage but was only 55% for deer in an early infection stage. The results indicate that the diagnostic sensitivity of tonsil biopsy for detecting early cases of CWD by immunohistochemistry is severely limited in white-tailed deer.

Technical Abstract: Chronic wasting disease (CWD) continues to spread in wild and farmed cervid populations. Early antemortem CWD testing of farmed cervids is of considerable interest to producers and regulatory agencies as a tool to combat this spread. The tissues accessible for antemortem sampling are limited and include biopsy of the tonsil and recto-anal mucosa-associated lymphoid tissue (RAMALT). The sensitivity to detect CWD by immunohistochemistry (IHC) —the regulatory gold standard—using biopsy samples of RAMALT from naturally infected white-tailed deer (WTD) has been determined by several studies. However, similar information is lacking for tonsil biopsy. In this study, two-bite tonsil biopsies from 79 naturally infected farmed WTD were used to determine the diagnostic sensitivity of tonsil IHC compared to the official CWD status based on results from the medial retropharyngeal lymph nodes and obex. IHC detection of CWD by tonsil biopsy was compared to the result and follicle metrics from the contralateral whole tonsil. The sensitivity of two-bite tonsil biopsy for detecting CWD by IHC was 72% overall. When the stage of infection was considered, the sensitivity was 92% for deer in late preclinical infection but only 55% for early preclinical infection. For deer with early preclinical infection, the sensitivity for deer homozygous for the prion protein gene (PRNP) coding for glycine at codon 96 (GG) was 66% but only 30% when heterozygous for the serine substitution (GS). The results indicate that the sensitivity of twobite tonsil biopsy in WTD, and consequently its potential utility as an antemortem diagnostic, is limited during early infection, especially in WTD heterozygous for the serine substitution at PRNP codon 96.