Discrimination between Scrapie and Bovine Spongiform Encephalopathy in Sheep by Molecular Size, Immunoreactivity, and Glycoprofile of Prion Protein

Thuring, C. M. A.; Erkens, J. H. F.; Jacobs, J.G.; Bossers, Alex; Keulen, L.J.M. van; Garssen, G.J.; Zijderveld, F.G. van; Ryder, Stephen; Groschup, Martin H.; Sweeney, T.; Langeveld, J.P.M.

doi:10.1128/jcm.42.3.972-980.2004

Cited by 110 publications

(106 citation statements)

References 45 publications

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“…In this regard, antibodies recognizing an epitope between residues 82 and 96 (ie, not detecting type 2) appeared straightforward tools to discriminate cases of animal-derived TSE strains. 38,[40][41][42][43] Using these antibodies on human CJD isolates, two groups of investigators recently found that all type 2 samples show at least some associated protein migrating similarly to PrP Sc type 1 and reached the conclusion Western blot profiles of PrP Sc in artificially mixed sCJD subtypes with little amount of PrP Sc type 1 (5% MM1 added to 95% MM2-cortical). Protease digestion was performed at different increasing PK concentrations; membranes were probed with 3F4 and the 'type 1-selective' antibody, 12B2.…”

Section: Discussionmentioning

confidence: 99%

A refined method for molecular typing reveals that co-occurrence of PrPSc types in Creutzfeldt–Jakob disease is not the rule

Notari

Capellari

Langeveld

et al. 2007

Laboratory Investigation

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Molecular typing in Creutzfeldt-Jakob disease (CJD) relies on the detection of distinct protease-resistant prion protein (PrP Sc ) core fragments, which differ in molecular mass or glycoform ratio. However, the definition and correct identification of CJD cases with a co-occurrence of PrP Sc types remains a challenge. With antibodies recognizing a linear epitope in the octapeptide repeat PrP region, supposed to distinguish between the two major PrP Sc isoforms (ie, types 1 and 2), it was recently shown that all type 2 cases display an associated band with a type 1 migration pattern, which led to the conclusion that multiple PrP Sc types regularly coexist in CJD. We studied brain samples from 53 sporadic CJD and 4 variant CJD subjects using a high-resolution electrophoresis, a wide range of proteinase K (PK) activities, the 'type 1-selective' antibody 12B2, and several unselective antibodies. We found that the type 1-like band detected by 12B2 in all CJD subtypes associated with PrP Sc type 2 is not a PK-resistant PrP Sc core but rather matches the physicochemical properties of partially cleaved fragments, which result from the several PK cleavage sites included in the N-terminal portion of PrP Sc . Furthermore, using gels with high resolution and a relatively high PK activity, we were able to increase the detection sensitivity of either type 1 or 2, when coexisting, to amount corresponding to 3-5% of the total PrP Sc signal (ie, weak band of one type/total PrP Sc ). Our results show that there are many pitfalls associated with the use of 'type 1 selective' antibodies for CJD typing studies and that co-occurrence of PrP Sc types in CJD is not the rule. The present results further validate the rationale basis of current CJD classification and the qualitative nature of molecular typing in CJD.

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Section: Discussionmentioning

confidence: 99%

A refined method for molecular typing reveals that co-occurrence of PrPSc types in Creutzfeldt–Jakob disease is not the rule

Notari

Capellari

Langeveld

et al. 2007

Laboratory Investigation

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“…For antigen retrieval, 4 mm tissue sections were immersed in formic acid for 5 min at 20 uC followed by autoclaving in 0.2 % citrate retrieval solution (pH 6.8) at 121 uC for 30 min. After washing in tap water and quenching in hydrogen peroxide (3 % in methanol) for 20 min, tissue sections were blocked with the MOM kit liquid protein concentrate solution (Vector Laboratories) at a dilution of 1 : 20 for 60 min and incubated overnight at 23 uC with 1 mg 2G11 ml 21 (Novus Biologicals) or 0.2 mg SAF84 ml 21 (SPI Bio), which are both mouse mAbs recognizing aa 153-158 (Thuring et al, 2004) and aa 166-172 (Jacobs et al, 2011) of ovine PrP, respectively. The subsequent steps of the IHC procedure were performed using an immunoperoxidase Elite ABC kit (Vector Laboratories).…”

Section: Methodsmentioning

confidence: 99%

Archival search for historical atypical scrapie in sheep reveals evidence for mixed infections

Chong

Kennedy

Goldmann

et al. 2015

Journal of General Virology

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“…Comparison of the signals obtained for both antibodies provides an easy and fast identifi cation of the BSE strain because binding of the anti-N terminal antibody is almost completely suppressed. This approach has been used to characterize unusual BSE cases in cattle (29) and naturally infected scrapie sheep (14,30).…”

mentioning

confidence: 99%

Rapid Typing of Transmissible Spongiform Encephalopathy Strains with Differential ELISA

Simon

Nugier

Morel

et al. 2008

Emerg. Infect. Dis.

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Discrimination between Scrapie and Bovine Spongiform Encephalopathy in Sheep by Molecular Size, Immunoreactivity, and Glycoprofile of Prion Protein

Cited by 110 publications

References 45 publications

A refined method for molecular typing reveals that co-occurrence of PrPSc types in Creutzfeldt–Jakob disease is not the rule

A refined method for molecular typing reveals that co-occurrence of PrPSc types in Creutzfeldt–Jakob disease is not the rule

Archival search for historical atypical scrapie in sheep reveals evidence for mixed infections

Rapid Typing of Transmissible Spongiform Encephalopathy Strains with Differential ELISA

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