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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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Analysis of goats isolates with ELISA typing test and immunoblot. Two experimental bovine spongiform encephalopathy (BSE) samples in goats analyzed by using the manual typing ELISA (A) gave ratios similar to those of experimental BSE in sheep. Results are the mean of 3 independent experiments. Experimental goat BSE and BSE-like field isolate Ch636 were analyzed by Western blot and compared with scrapie goat isolates (B). Lane 1, untreated negative brain homogenate. Lane 2, proteinase K (PK)–treated negative brain homogenate. Lanes 3–9, PK-treated positive isolates: French scrapie isolate 99–1316 (lane 3); experimental ovine BSE 397 BS (lane 4); experimental BSE goat 2 (lane 5); French goat isolate Ch 636 (campaign 2002) (lane 6); French scrapie goat isolates Ch517 and Ch519 (campaign 2002) (lanes 7 and 8, respectively); Norwegian scrapie isolate (Lavik, lane 9). MW, molecular weight.
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Figure 4: Analysis of goats isolates with ELISA typing test and immunoblot. Two experimental bovine spongiform encephalopathy (BSE) samples in goats analyzed by using the manual typing ELISA (A) gave ratios similar to those of experimental BSE in sheep. Results are the mean of 3 independent experiments. Experimental goat BSE and BSE-like field isolate Ch636 were analyzed by Western blot and compared with scrapie goat isolates (B). Lane 1, untreated negative brain homogenate. Lane 2, proteinase K (PK)–treated negative brain homogenate. Lanes 3–9, PK-treated positive isolates: French scrapie isolate 99–1316 (lane 3); experimental ovine BSE 397 BS (lane 4); experimental BSE goat 2 (lane 5); French goat isolate Ch 636 (campaign 2002) (lane 6); French scrapie goat isolates Ch517 and Ch519 (campaign 2002) (lanes 7 and 8, respectively); Norwegian scrapie isolate (Lavik, lane 9). MW, molecular weight.

Mentions: As shown in Figure 3, only 10 samples (3.8%) provided ratios compatible with experimental BSE (normalized ratio 0.7–1.3), and 28 provided ratios superior to experimental BSE in sheep (normalized ratio >1.3–10.5, Figure 3). Twenty-nine samples, as well as 4 of the 10 samples compatible with experimental BSE and 9 samples with low levels of PrPres, were previously identified as atypical scrapie (open bars, Figure 3). Among the samples that yielded ratios compatible with experimental BSE in sheep, 1 goat isolate, Ch636, was extensively studied because of its BSE-like profile in 3 different Western blot techniques (1). As shown in Figure 3, this sample had a normalized A/A′ratio close to 1, very similar to that for experimental goat BSE (Figure 4, panel A, Table 2). This result is confirmed by the migration pattern of the nonglycosylated band (Figure 4, panel B, lane 6), which appears very similar to that of experimental BSE in sheep (Figure 4, panel B, lane 4) and of experimental goat BSE (lane 5), and different from that of French scrapie goat isolates (lanes 7 and 8).


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)

Analysis of goats isolates with ELISA typing test and immunoblot. Two experimental bovine spongiform encephalopathy (BSE) samples in goats analyzed by using the manual typing ELISA (A) gave ratios similar to those of experimental BSE in sheep. Results are the mean of 3 independent experiments. Experimental goat BSE and BSE-like field isolate Ch636 were analyzed by Western blot and compared with scrapie goat isolates (B). Lane 1, untreated negative brain homogenate. Lane 2, proteinase K (PK)–treated negative brain homogenate. Lanes 3–9, PK-treated positive isolates: French scrapie isolate 99–1316 (lane 3); experimental ovine BSE 397 BS (lane 4); experimental BSE goat 2 (lane 5); French goat isolate Ch 636 (campaign 2002) (lane 6); French scrapie goat isolates Ch517 and Ch519 (campaign 2002) (lanes 7 and 8, respectively); Norwegian scrapie isolate (Lavik, lane 9). MW, molecular weight.
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Related In: Results  -  Collection

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Figure 4: Analysis of goats isolates with ELISA typing test and immunoblot. Two experimental bovine spongiform encephalopathy (BSE) samples in goats analyzed by using the manual typing ELISA (A) gave ratios similar to those of experimental BSE in sheep. Results are the mean of 3 independent experiments. Experimental goat BSE and BSE-like field isolate Ch636 were analyzed by Western blot and compared with scrapie goat isolates (B). Lane 1, untreated negative brain homogenate. Lane 2, proteinase K (PK)–treated negative brain homogenate. Lanes 3–9, PK-treated positive isolates: French scrapie isolate 99–1316 (lane 3); experimental ovine BSE 397 BS (lane 4); experimental BSE goat 2 (lane 5); French goat isolate Ch 636 (campaign 2002) (lane 6); French scrapie goat isolates Ch517 and Ch519 (campaign 2002) (lanes 7 and 8, respectively); Norwegian scrapie isolate (Lavik, lane 9). MW, molecular weight.
Mentions: As shown in Figure 3, only 10 samples (3.8%) provided ratios compatible with experimental BSE (normalized ratio 0.7–1.3), and 28 provided ratios superior to experimental BSE in sheep (normalized ratio >1.3–10.5, Figure 3). Twenty-nine samples, as well as 4 of the 10 samples compatible with experimental BSE and 9 samples with low levels of PrPres, were previously identified as atypical scrapie (open bars, Figure 3). Among the samples that yielded ratios compatible with experimental BSE in sheep, 1 goat isolate, Ch636, was extensively studied because of its BSE-like profile in 3 different Western blot techniques (1). As shown in Figure 3, this sample had a normalized A/A′ratio close to 1, very similar to that for experimental goat BSE (Figure 4, panel A, Table 2). This result is confirmed by the migration pattern of the nonglycosylated band (Figure 4, panel B, lane 6), which appears very similar to that of experimental BSE in sheep (Figure 4, panel B, lane 4) and of experimental goat BSE (lane 5), and different from that of French scrapie goat isolates (lanes 7 and 8).

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