|McGarvey, Jeffery - Jeff|
Submitted to: Journal of Immunological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/15/2018
Publication Date: 2/17/2018
Citation: Hnasko, R.M., Lin, A.V., McGarvey, J.A., Stanker, L.H. 2018. Enhanced detection of infectious prions by direct ELISA from the brains of asymptomatic animals using DRM2-118 monoclonal antibody and Gdn-HCl. Journal of Immunological Methods. 456:38-43. https://doi.org/10.1016/j.jim.2018.02.010.
Interpretive Summary: We have developed novel tools and methods that improve the detection of infectious prions by immunoassay. Improved sample preparation methods along with simple immunoassay modifications result in increased prion detection allowing for disease confirmation before the onset of disease symptoms or other pathological changes. The use of these methodologies will enhance ongoing prion disease surveillance efforts to improve animal health and the safety of agricultural products.
Technical Abstract: In this report we describe improved methods for the detection of infectious prions by immunoassay for the diagnosis of transmissible spongiform encephalopathies (TSEs) from asymptomatic animals. Tissue samples obtained as part of ongoing TSE surveillance efforts are often unsuitable for histopathological assessment or lack defined pathological characteristics of late stage disease. The etiological agents of TSEs are abnormal infectious prions (PrPSc) and they can be conclusively identified by immunoassay via their resistance to proteolytic degradation. In the absence of histopathological detail from suspect animals the confirmation of TSE disease requires sensitive immunoassays for the detection of infectious prion proteins to adequately determine the incidence of disease. Our data demonstrates enhanced prion detection from brains of asymptomatic animals by immunoassay using prion enrichment with lipid rafts along with a modified immunoassay that uses the chaotropic agent guanidine-HCl to increase antibody epitope binding.