title: Failure to detect abnormal prion protein and scrapie-associated fibrils 6 wk after intracerebral inoculation of genetically susceptible sheep with scrapie agent.
abstract: Detection of the scrapie-associated protease-resistant prion protein (PrPres) in sheep brains in the early phase after intracerebral inoculation of the scrapie agent has not been documented. Fourteen 4-mo-old, genetically susceptible lambs (QQ homozygous at codon 171 of the PrP gene) were obtained for this study. Twelve lambs were inoculated intracerebrally with a brain suspension from sheep naturally affected with scrapie, and 2 served as uninoculated controls. Two inoculated animals were euthanized at each of 6 times postinoculation (1 h to 6 wk), and their brains were collected for histopathological study, for detection of PrPres by the Western blot technique and an immunohistochemical (IHC) method, and for the detection of scrapie-associated fibrils (SAF) by negatively stained electron microscopy (EM). Microscopic lesions associated with introduction of the inoculum were seen in the brains of inoculated animals at all 6 times. However, both the Western blot and IHC techniques did not detect PrPres after the initial 3 d postinoculation, nor did EM detect SAF in any of the samples. From these findings, it is presumed that until host amplification has occurred, the concentration of PrPres in inoculum is insufficient for detection by currently available techniques.
authors.affiliation: National Animal Disease Center, Agricultural Research Service, US Department of Agriculture, Ames, Iowa 50010, USA. ahamir@nadc.ars.usda.gov
authors.forename: Amir N
authors.initials: AN
authors.lastname: Hamir
authors.forename: Janice M
authors.initials: JM
authors.lastname: Miller
authors.forename: Mick J
authors.initials: MJ
authors.lastname: Stack
authors.forename: Melanie J
authors.initials: MJ
authors.lastname: Chaplin
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western / Q000662:veterinary; D001921:Brain / Q000648:ultrastructure; D005260:Female; D020022:Genetic Predisposition to Disease; D015151:Immunoblotting / Q000662:veterinary; D007267:Injections / Q000662:veterinary; D008297:Male; D018622:PrP 27-30 Protein / Q000302:isolation & purification / Q000648:ultrastructure; D018620:PrPSc Proteins / Q000302:isolation & purification* / Q000648:ultrastructure*; D011237:Predictive Value of Tests; D012608:Scrapie / Q000175:diagnosis* / Q000235:genetics*; D012756:Sheep; D013997:Time Factors
chemical_list: D018620:PrPSc Proteins; D018622:PrP 27-30 Protein