title: Molecular discrimination of atypical bovine spongiform encephalopathy strains from a geographical region spanning a wide area in Europe.
abstract: Transmissible spongiform encephalopathy strains can be differentiated by their behavior in bioassays and by molecular analyses of the disease-associated prion protein (PrP) in a posttranslationally transformed conformation (PrPSc). Until recently, isolates from cases of bovine spongiform encephalopathy (BSE) appeared to be very homogeneous. However, a limited number of atypical BSE isolates have recently been identified upon analyses of the disease-associated proteinase K (PK) resistance-associated moiety of PrPSc (PrPres), suggesting the existence of at least two additional BSE PrPres variants. These are defined here as the H type and the L type, according to the higher and lower positions of the nonglycosylated PrPres band in Western blots, respectively, compared to the position of the band in classical BSE (C-type) isolates. These molecular PrPres variants, which originated from six different European countries, were investigated together. In addition to the migration properties and glycosylation profiles (glycoprofiles), the H- and L-type isolates exhibited enhanced PK sensitivities at pH 8 compared to those of the C-type isolates. Moreover, H-type BSE isolates exhibited differences in the binding of antibodies specific for N- and more C-terminal PrP regions and principally contained two aglycosylated PrPres moieties which can both be glycosylated and which is thus indicative of the existence of two PrPres populations or intermediate cleavage sites. These properties appear to be consistent within each BSE type and independent of the geographical origin, suggesting the existence of different BSE strains in cattle. The choice of three antibodies and the application of two pHs during the digestion of brain homogenates provide practical and diverse tools for the discriminative detection of these three molecular BSE types and might assist with the recognition of other variants.
authors: Central Institute for Animal Disease Control (CIDC-Lelystad), 8203 AA 2004, Lelystad. The Netherlands.
authors: Jorg G
authors: JG
authors: Jacobs
authors: Jan P M
authors: JP
authors: Langeveld
authors: Anne-Gaëlle
authors: AG
authors: Biacabe
authors: Pier-Luigi
authors: PL
authors: Acutis
authors: Miroslaw P
authors: MP
authors: Polak
authors: Dolores
authors: D
authors: Gavier-Widen
authors: Anne
authors: A
authors: Buschmann
authors: Maria
authors: M
authors: Caramelli
authors: Cristina
authors: C
authors: Casalone
authors: Maria
authors: M
authors: Mazza
authors: Martin
authors: M
authors: Groschup
authors: Jo H F
authors: JH
authors: Erkens
authors: Aart
authors: A
authors: Davidse
authors: Fred G
authors: FG
authors: van Zijderveld
authors: Thierry
authors: T
authors: Baron
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D015153:Blotting, Western; D001933:Brain Stem / Q000378:metabolism*; D002417:Cattle; D016643:Encephalopathy, Bovine Spongiform / Q000453:epidemiology* / Q000378:metabolism*; D019286:Endopeptidase K / Q000378:metabolism / Q000494:pharmacology; D005060:Europe / Q000453:epidemiology; D014644:Genetic Variation; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D018620:PrPSc Proteins / Q000145:classification* / Q000235:genetics* / Q000378:metabolism; D011328:Prions / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D018620:PrPSc Proteins; D011328:Prions; D019286:Endopeptidase K