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Rapid typing of transmissible spongiform encephalopathy strains with differential ELISA.

Simon S, Nugier J, Morel N, Boutal H, Créminon C, Benestad SL, Andréoletti O, Lantier F, Bilheude JM, Feyssaguet M, Biacabe AG, Baron T, Grassi J - Emerging Infect. Dis. (2008)

Bottom Line: Based on the particular biochemical properties of the abnormal prion protein (PrPsc) associated with BSE, and particularly the increased degradation induced by proteinase K in the N terminal part of PrPsc, we have developed a rapid ELISA designed to distinguish BSE from other scrapie strains.This assay clearly discriminates experimental ovine BSE from other scrapie strains and was used to screen 260 transmissible spongiform encephalopathy (TSE)-infected small ruminant samples identified by the French active surveillance network (2002/2003).In this context, this test has helped to identify the first case of natural BSE in a goat and can be used to classify TSE isolates based on the proteinase K sensitivity of PrPsc.

View Article: PubMed Central - PubMed

Affiliation: Commissariat à l'Energie Atomique, Gif-sur-Yvette, France.

ABSTRACT
The bovine spongiform encephalopathy (BSE) agent has been transmitted to humans, leading to variant Creutzfeldt-Jakob disease. Sheep and goats can be experimentally infected by BSE and have been potentially exposed to natural BSE; however, whether BSE can be transmitted to small ruminants is not known. Based on the particular biochemical properties of the abnormal prion protein (PrPsc) associated with BSE, and particularly the increased degradation induced by proteinase K in the N terminal part of PrPsc, we have developed a rapid ELISA designed to distinguish BSE from other scrapie strains. This assay clearly discriminates experimental ovine BSE from other scrapie strains and was used to screen 260 transmissible spongiform encephalopathy (TSE)-infected small ruminant samples identified by the French active surveillance network (2002/2003). In this context, this test has helped to identify the first case of natural BSE in a goat and can be used to classify TSE isolates based on the proteinase K sensitivity of PrPsc.

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Related in: MedlinePlus

Principle of the 2-site immunometric typing assay. A) EIA screening test. In these conditions (Bio-Rad, Hercules, CA, USA), after mild proteinase K (PK) digestion, denatured PK-resistant prion protein (PrPres) is captured by the solid phase–immobilized antibody SAF34, recognizing the octarepeat region, and shown by the Bar224 tracer antibody, directed against the core of the protein. B) EIA typing test. In this test, the PrPres-containing sample is treated in 2 sets of PK conditions. In the first set of conditions (PK treatment in A reagent, Bio-Rad screening condition), the octarepeat region is maintained in scrapie- and bovine spongiform encephalopathy (BSE)–associated PrPres; in the second set of conditions (high PK concentration in A′reagent), BSE- and labile strain–associated PrPres is more sensitive to PK digestion than the classic PrPres associated with scrapie strains. Calculation of the ratio in the 2 conditions differentiates BSE and labile strains from classic scrapie strains. ab, antibody.
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Figure 1: Principle of the 2-site immunometric typing assay. A) EIA screening test. In these conditions (Bio-Rad, Hercules, CA, USA), after mild proteinase K (PK) digestion, denatured PK-resistant prion protein (PrPres) is captured by the solid phase–immobilized antibody SAF34, recognizing the octarepeat region, and shown by the Bar224 tracer antibody, directed against the core of the protein. B) EIA typing test. In this test, the PrPres-containing sample is treated in 2 sets of PK conditions. In the first set of conditions (PK treatment in A reagent, Bio-Rad screening condition), the octarepeat region is maintained in scrapie- and bovine spongiform encephalopathy (BSE)–associated PrPres; in the second set of conditions (high PK concentration in A′reagent), BSE- and labile strain–associated PrPres is more sensitive to PK digestion than the classic PrPres associated with scrapie strains. Calculation of the ratio in the 2 conditions differentiates BSE and labile strains from classic scrapie strains. ab, antibody.

Mentions: The capture antibody recognizes an amino terminal epitope, while the tracer antibody binds to the core of the protein (Figure 1, panel A). In the first set of conditions, the PK digestion is performed in a control medium (mixture of detergents and chaotropic agents) to preserve the N-terminal epitope (Figure 1, panels A and B, PK treatment in A conditions). By varying the conditions of PK treatment, PrPsc associated with the BSE strain appears more sensitive to PK than most of the other prion strains. Conditions of PK treatment can thus be defined for selectively deleting the epitope recognized by the capture antibody in BSE strain while preserving it in most scrapie strains (Figure 1, panel B, PK in A′conditions), in agreement with previous reports (5,10,23,28).


Rapid typing of transmissible spongiform encephalopathy strains with differential ELISA.

Simon S, Nugier J, Morel N, Boutal H, Créminon C, Benestad SL, Andréoletti O, Lantier F, Bilheude JM, Feyssaguet M, Biacabe AG, Baron T, Grassi J - Emerging Infect. Dis. (2008)

Principle of the 2-site immunometric typing assay. A) EIA screening test. In these conditions (Bio-Rad, Hercules, CA, USA), after mild proteinase K (PK) digestion, denatured PK-resistant prion protein (PrPres) is captured by the solid phase–immobilized antibody SAF34, recognizing the octarepeat region, and shown by the Bar224 tracer antibody, directed against the core of the protein. B) EIA typing test. In this test, the PrPres-containing sample is treated in 2 sets of PK conditions. In the first set of conditions (PK treatment in A reagent, Bio-Rad screening condition), the octarepeat region is maintained in scrapie- and bovine spongiform encephalopathy (BSE)–associated PrPres; in the second set of conditions (high PK concentration in A′reagent), BSE- and labile strain–associated PrPres is more sensitive to PK digestion than the classic PrPres associated with scrapie strains. Calculation of the ratio in the 2 conditions differentiates BSE and labile strains from classic scrapie strains. ab, antibody.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2570920&req=5

Figure 1: Principle of the 2-site immunometric typing assay. A) EIA screening test. In these conditions (Bio-Rad, Hercules, CA, USA), after mild proteinase K (PK) digestion, denatured PK-resistant prion protein (PrPres) is captured by the solid phase–immobilized antibody SAF34, recognizing the octarepeat region, and shown by the Bar224 tracer antibody, directed against the core of the protein. B) EIA typing test. In this test, the PrPres-containing sample is treated in 2 sets of PK conditions. In the first set of conditions (PK treatment in A reagent, Bio-Rad screening condition), the octarepeat region is maintained in scrapie- and bovine spongiform encephalopathy (BSE)–associated PrPres; in the second set of conditions (high PK concentration in A′reagent), BSE- and labile strain–associated PrPres is more sensitive to PK digestion than the classic PrPres associated with scrapie strains. Calculation of the ratio in the 2 conditions differentiates BSE and labile strains from classic scrapie strains. ab, antibody.
Mentions: The capture antibody recognizes an amino terminal epitope, while the tracer antibody binds to the core of the protein (Figure 1, panel A). In the first set of conditions, the PK digestion is performed in a control medium (mixture of detergents and chaotropic agents) to preserve the N-terminal epitope (Figure 1, panels A and B, PK treatment in A conditions). By varying the conditions of PK treatment, PrPsc associated with the BSE strain appears more sensitive to PK than most of the other prion strains. Conditions of PK treatment can thus be defined for selectively deleting the epitope recognized by the capture antibody in BSE strain while preserving it in most scrapie strains (Figure 1, panel B, PK in A′conditions), in agreement with previous reports (5,10,23,28).

Bottom Line: Based on the particular biochemical properties of the abnormal prion protein (PrPsc) associated with BSE, and particularly the increased degradation induced by proteinase K in the N terminal part of PrPsc, we have developed a rapid ELISA designed to distinguish BSE from other scrapie strains.This assay clearly discriminates experimental ovine BSE from other scrapie strains and was used to screen 260 transmissible spongiform encephalopathy (TSE)-infected small ruminant samples identified by the French active surveillance network (2002/2003).In this context, this test has helped to identify the first case of natural BSE in a goat and can be used to classify TSE isolates based on the proteinase K sensitivity of PrPsc.

View Article: PubMed Central - PubMed

Affiliation: Commissariat à l'Energie Atomique, Gif-sur-Yvette, France.

ABSTRACT
The bovine spongiform encephalopathy (BSE) agent has been transmitted to humans, leading to variant Creutzfeldt-Jakob disease. Sheep and goats can be experimentally infected by BSE and have been potentially exposed to natural BSE; however, whether BSE can be transmitted to small ruminants is not known. Based on the particular biochemical properties of the abnormal prion protein (PrPsc) associated with BSE, and particularly the increased degradation induced by proteinase K in the N terminal part of PrPsc, we have developed a rapid ELISA designed to distinguish BSE from other scrapie strains. This assay clearly discriminates experimental ovine BSE from other scrapie strains and was used to screen 260 transmissible spongiform encephalopathy (TSE)-infected small ruminant samples identified by the French active surveillance network (2002/2003). In this context, this test has helped to identify the first case of natural BSE in a goat and can be used to classify TSE isolates based on the proteinase K sensitivity of PrPsc.

Show MeSH
Related in: MedlinePlus