A single step multiplex immunofluorometric assay for differential diagnosis of BSE and scrapie

Sheep prions with molecular properties intermediate between classical scrapie, BSE and CH1641–scrapie

“…Diagnosis targeted on the N-terminal proteolytic processing of PrP Sc is being an effective tool in discriminating TSE strains either without the use of protease digestion as in immunohistochemistry (IHC) [11][12][13] or with additional digestion using proteinase K as performed in Western blotting, bead-based multiplex assays and ELISA assays. [14][15][16][17][18] Of these techniques, Western blotting is able to both recognize PrP and provide essential information on different molecular PrP Sc properties adding to the phenotypical palette of parameters for TSE strain typing. In this way, several types have been discovered in ruminants.…”

“…8,[12][13][14][15][16][22][23][24][25][26][27][28][29][30] However, at least in small ruminants there are potentially several phenotypes present in the same host which pose basic questions about the nature of the agent, the source of the BSE epidemic and how to recognize the presence of BSE. [31][32][33][34][35][36] Recently we introduced 2 unique and powerful multiplex TSE differentiation non-enzymatic assays using fluorescence detection: one bead-based assay for TSE-typing in sheep 18,37 and one probing technique for Western blots. 38 These 2 techniques were effective to differentiate in brain homogenates between classical scrapie, CH1641, BSE, and Nor98/atypical scrapie in one test using 3 antibodies in a single mixture.…”

Differentiation of ruminant transmissible spongiform encephalopathy isolate types, including bovine spongiform encephalopathy and CH1641 scrapie

“…Until now, several sections or blots are needed to evaluate partial removal of epitopes and variations in PrP glycoprofile. In Western blots (WB), BSE in sheep can be discriminated from classical scrapie in sheep (Baron et al, 2000;Hill et al, 1998;Nonno et al, 2003;Stack et al, 2002;Tang et al, 2010;Thuring et al, 2004) based on differences in PrP res length in the region of ovine PrP residues 80-99. CH1641 scrapie is an unusual and rare form of sheep scrapie (Foster & Dickinson, 1988), which has N-terminal PrP res features resembling those of BSE, while its PrP res glycosylation characteristics seem slightly different; furthermore its pathological and transmission properties appeared deviant from BSE (Baron et al, 2000;Hope et al, 1999;Jeffrey et al, 2006a;Somerville et al, 1997;Stack et al, 2002).…”

Prion Protein-Specific Antibodies that Detect Multiple TSE Agents with High Sensitivity

“…This is of both functional and biological significance, since data emanating from the use of antibodies with different epitopes within PrP have yielded significant advancements in prion biology relating to: diagnostic regimes for animal TSE disease (using 6H4 [22], [24], [51], [52] or human TSEs (using KG9 singly or in combination with other antibodies i.e. 3F4, ICMS35, BG4 [53]–[56]); immunological characterisation of pathology associated with established and emerging TSEs, such as immunophenotyping of atypical scrapie using an array of antibodies [2] or identification of the shared characteristic of Nor98 scrapie and human Gerstmann-Straussler-Scheinker disease (GSS) [6]; the use of sequences in the prion protein which are species specific (using 3F4 or antisera raised against synthetic peptides [14], [25], [57]); the discrimination of TSE strains in large and small animal models, for example natural scrapie, BSE and CH1641 scrapie (using P4 and 66.94ba [58], 6H4/P4 [59], P4 [60]), using triplex immunostaining with L42, 12B2 and SAF84 [61] and using multiple antibody panels [5], [41], [62]–[67]; the characterisation and classification of human prion disease [68]–[71]; the characterisation of structural elements within PrP, such as deciphering accessibility and exposure of epitopes following the conversion of PrP C to PrP Sc or those that are PrP Sc -specific i.e.…”