title: Detection and Quantification of CWD Prions in Fixed Paraffin Embedded Tissues by Real-Time Quaking-Induced Conversion.
abstract: Traditional diagnostic detection of chronic wasting disease (CWD) relies on immunodetection of misfolded CWD prion protein (PrP(CWD)) by western blotting, ELISA, or immunohistochemistry (IHC). These techniques require separate sample collections (frozen and fixed) which may result in discrepancies due to variation in prion tissue distribution and assay sensitivities that limit detection especially in early and subclinical infections. Here, we harness the power of real-time quaking induced conversion (RT-QuIC) to amplify, detect, and quantify prion amyloid seeding activity in fixed paraffin-embedded (FPE) tissue sections. We show that FPE RT-QuIC has greater detection sensitivity than IHC in tissues with low PrP(CWD) burdens, including those that are IHC-negative. We also employ amyloid formation kinetics to yield a semi-quantitative estimate of prion concentration in a given FPE tissue. We report that FPE RT-QuIC has the ability to enhance diagnostic and investigative detection of disease-associated PrP(RES) in prion, and potentially other, protein misfolding disease states.
authors.affiliation: Prion Research Center, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins CO 80523, USA.
authors.forename: Clare E
authors.initials: CE
authors.lastname: Hoover
authors.affiliation: Prion Research Center, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins CO 80523, USA.
authors.forename: Kristen A
authors.initials: KA
authors.lastname: Davenport
authors.affiliation: Prion Research Center, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins CO 80523, USA.
authors.forename: Davin M
authors.initials: DM
authors.lastname: Henderson
authors.affiliation: Prion Research Center, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins CO 80523, USA.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Pulscher
authors.affiliation: Prion Research Center, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins CO 80523, USA.
authors.forename: Candace K
authors.initials: CK
authors.lastname: Mathiason
authors.affiliation: Prion Research Center, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins CO 80523, USA.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Zabel
authors.affiliation: Prion Research Center, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins CO 80523, USA.
authors.forename: Edward A
authors.initials: EA
authors.lastname: Hoover
mesh_terms: D000818:Animals; D006651:Histocytochemistry / Q000379:methods*; D011328:Prions / Q000032:analysis*; D012680:Sensitivity and Specificity; D013997:Time Factors; D034081:Wasting Disease, Chronic / Q000175:diagnosis*
chemical_list: D011328:Prions