Approaches to Scrapie diagnosis by applying immunohistochemistry and rapid tests on central nervous and lymphoreticular systems

Monleón, Eva; Monzón, Marta; Hortells, Paloma; Bolea, Rosa; Acín, Cristina; Vargas, Francisco Suso; Badiola, Juan José

doi:10.1016/j.jviromet.2005.01.013

Cited by 18 publications

(16 citation statements)

References 33 publications

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“…In this context, the ability of early and safe detection is vital for scrapie diagnosis to have real value. Particularly in animals destined for human consumption, where there is an additional requirement to determine the presence of the prion, hence our choice to use CNS from apparently healthy and clinically asymptomatic slaughter house goats that are destined for consumption [6,16].…”

Section: Discussionmentioning

confidence: 99%

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Differential Diagnosis of Scrapie in Post Mortem Goats by immunohistochemistry

Farías¹,

Frohlich²,

Sanhueza³

et al. 2017

JDVAR

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Scrapie is a neurodegenerative disease of slow and lethal progression affecting sheep, goats and mouflons, which is part of a group of pathologies produced by prions or transmissible spongiform diseases (TSEs), these affect both humans and animals. Based on the fact that immunohistochemistry (IHC) is the internationally recognized method for the diagnosis of classical scrapie in goats, a species considered as a reservoir for the disease that may be zoonotic, we aimed to detect and differentiate it from other lesions or pathologies of the central nervous system. Chile in relation to bovine spongiform encephalopathy (BSE), is classified as a disease-free country. Scrapie is actively monitored for the sheep, but passively for the caprine species, which is why this study was carried out in slaughtered goats in the central north of the country (IV Region), to provide information to the surveillance system for this disease. Fifty brains of goats older than 2 years were used, regardless of race or gender. The obex was taken from each of them, two serial histological sections were obtained, one used for the staining of Hematoxylin and Eosin (H/E) (50 cuts), which allowed us to observe lesions and to evaluate the aptitude of the samples. The other one was used for immunohistochemistry (IHC) (50 cuts), which allows to locate prions in situ, using a commercial kit and the technique described by Manning et al. [1]. For a correct diagnosis, 4 controls were used, 2 positive (1 ovine and 1 caprine) and 2 negative (1 ovine and 1 caprine), provided by an International Reference Center. Analysis of the 50 H/E histological sections indicated that all were eligible for the IHC technique, and non-specific lesions (38/50) were found, such as focal haemorrhage, perivascular infiltrate and pigments, all of which are not attributable to prions. With the IHC technique, all the homologous histological sections of the samples and the negative controls were classified as negative to classic scrapie, due to the fact that they did not present immunostaining characterized by a red granular precipitate, which was present in the positive controls used. The results obtained in this work validate the application of the IHC technique at the level of obex, as a method of diagnosis of classic goat scrapie in the country.

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Section: Discussionmentioning

confidence: 99%

“…In other species such as cattle or sheep, other diagnostic tests, such as the western blot or the ELISA test, these ones have not been widely used in goats [1,6].…”

Section: Introductionmentioning

confidence: 99%

Differential Diagnosis of Scrapie in Post Mortem Goats by immunohistochemistry

Farías¹,

Frohlich²,

Sanhueza³

et al. 2017

JDVAR

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“…Historically, scrapie has been considered nonpathogenic for humans (4), but because of the possibility of natural BSE transmission, which has been evidenced in a French goat, the European Union decided that TSE-affected sheep and goats must be kept out of the food chain to avoid possible presence of BSE (5).…”

Section: Introductionmentioning

confidence: 99%

“…Histologically, 2 types of characteristics have been described: 1) PrP Sc deposition in central nervous and lymphoreticular systems and 2) histopathologic lesions in specific areas of the central nervous system, mainly including vacuolation of neuronal perikarya and grey matter neuropils and also neuronal degeneration and neuronal loss (especially Bdark[ neurons), an increase of reactive glial cells (predominantly astrocytes), and amyloidosis (4,6,7).…”

Section: Introductionmentioning

confidence: 99%

Changes in the Expression Pattern of the Nitrergic System of Ovine Cerebellum Affected by Scrapie

Fernández

Serrano

Rodrigo

et al. 2007

Journal of Neuropathology and Experimental Neurology

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The constitutive and inducible isoforms of nitric oxide synthase (NOS) and the end-product of nitration, nitrotyrosine, were analyzed by immunohistochemistry, Western blotting, and enzymatic activity in sheep at different stages of the prion disease, scrapie. Four groups were studied: 1) nonaffected (control), 2) preclinical, 3) clinical, and 4) terminal. Constitutive neuronal NOS (nNOS) was the most abundant isoform present in cerebellar neurons of the sheep. Expression of nNOS increased in preclinical animals but diminished in the late stages of the disease. The Purkinje cells that usually are not immunoreactive for this protein became immunopositive in the clinical phase. In unaffected sheep, the inducible isoform (iNOS) was slightly positive in the Purkinje cells. As the disease progressed, the immunoreactivity of Purkinje neurons for iNOS increased. At the final stages, numerous iNOS-positive microglial cells were found in the molecular layer. There was a basal level of protein nitration in the cerebellum of unaffected sheep, especially in the molecular layer. As the disease progressed, the distal prolongations of the Purkinje cells and the astroglia became immunoreactive for nitrotyrosine. Our results suggest that the nitrergic system reacts to the progression of spongiform diseases and may be part of their pathogenesis mechanism.

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“…Immunohistochemistry was done using the L42 f antibody against PrP sc as described previously. 3 Briefly, the applied pretreatments included immersion in formic acid for 15 min, PK e treatment for 15 min at 37uC (4 mg/ml), and hydrated autoclaving. After incubating with the primary antibody (1:500 dilution for 30 min), a polymer-bound secondary antibody visualization system was applied, g and 3,39-diaminobenzidine was used as the chromogen substrate.…”

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confidence: 99%

Rapid Chromatographic Immunoassay Study of Brain PrP^sc Distribution in Classical Scrapie

Bolea

Vidal²,

Marín

et al. 2010

J VET Diagn Invest

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In the current study, a rapid chromatographic immunoassay submitted for registration in Europe was used to analyze PrP(sc) in 13 different areas of brain from 10 confirmed classical scrapie cases. The levels of PrP(sc) in the different areas of brain were plotted to draw a brain PrP(sc) distribution curve. This curve was compared with the brain PrP(sc) distribution curve obtained from immunoblotting and immunohistochemistry tests on the same samples. The distribution of PrP(sc) in different areas of the brain was similar, irrespective of the test applied, indicating that any of the 3 tests could be used for the characterization of classical cases of scrapie.

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Approaches to Scrapie diagnosis by applying immunohistochemistry and rapid tests on central nervous and lymphoreticular systems

Cited by 18 publications

References 33 publications

Differential Diagnosis of Scrapie in Post Mortem Goats by immunohistochemistry

Differential Diagnosis of Scrapie in Post Mortem Goats by immunohistochemistry

Changes in the Expression Pattern of the Nitrergic System of Ovine Cerebellum Affected by Scrapie

Rapid Chromatographic Immunoassay Study of Brain PrP^sc Distribution in Classical Scrapie

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Approaches to Scrapie diagnosis by applying immunohistochemistry and rapid tests on central nervous and lymphoreticular systems

Cited by 18 publications

References 33 publications

Differential Diagnosis of Scrapie in Post Mortem Goats by immunohistochemistry

Differential Diagnosis of Scrapie in Post Mortem Goats by immunohistochemistry

Changes in the Expression Pattern of the Nitrergic System of Ovine Cerebellum Affected by Scrapie

Rapid Chromatographic Immunoassay Study of Brain PrPsc Distribution in Classical Scrapie

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Rapid Chromatographic Immunoassay Study of Brain PrP^sc Distribution in Classical Scrapie