Postmortem diagnosis of preclinical and clinical scrapie in sheep by the detection of disease‐associated PrP in their rectal mucosa

González, Lorenzo; Dagleish, Mark P.; Bellworthy, S. J.; Sisó, Sílvia; Stack, M.J.; Chaplin, Melanie J.; Davis, L.A.; Hawkins, S. A. C.; Hughes, John T.; Jeffrey, Martin

doi:10.1136/vr.158.10.325

Cited by 66 publications

(73 citation statements)

References 30 publications

(4 reference statements)

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“…Previous work had suggested that examination of LRS tissues in addition to brain provides better estimates of scrapie infection in sheep [13,25,28]. Our results are a noticeable example that this is also the case in goats and that, in agreement with those earlier sheep studies, the medial retropharyngeal lymph node would be the sample of choice, together with the obex, for post-mortem surveillance.…”

Section: Implications For Surveillance Of Goat Tsesupporting

confidence: 88%

“…Post-mortem detection of PrP d in the RAMALT was achieved in a low percentage [13]), and similar ones in chronic wasting disease of cervids [21,33]. Whether this is a feature of goat scrapie in general or a peculiarity of this outbreak is unknown but, in any case, the frequency of detection of PrP d in the RAMALT rises to 84.0% when goats with widespread LRS involvement -but still at preclinical stagesare considered.…”

Section: Implications For Surveillance Of Goat Tsementioning

confidence: 77%

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High prevalence of scrapie in a dairy goat herd: tissue distribution of disease-associated PrP and effect ofPRNPgenotype and age

González¹,

Martin²,

Sisó³

et al. 2009

Vet. Res.

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-Following a severe outbreak of clinical scrapie in 2006-2007, a large dairy goat herd was culled and 200 animals were selected for post-mortem examinations in order to ascertain the prevalence of infection, the effect of age, breed and PRNP genotype on the susceptibility to scrapie, the tissue distribution of diseaseassociated PrP (PrP d ), and the comparative efficiency of different diagnostic methods. As determined by immunohistochemical (IHC) examinations with Bar224 PrP antibody, the prevalence of preclinical infection was very high (72/200; 36.0%), with most infected animals being positive for PrP d in lymphoreticular system (LRS) tissues (68/72; 94.4%) compared to those that were positive in brain samples (38/72; 52.8%). The retropharyngeal lymph node and the palatine tonsil showed the highest frequency of PrP d accumulation (87.3% and 84.5%, respectively), while the recto-anal mucosa-associated lymphoid tissue (RAMALT) was positive in only 30 (41.7%) of the infected goats. However, the efficiency of rectal and palatine tonsil biopsies taken shortly before necropsy was similar. The probability of brain and RAMALT being positive directly correlated with the spread of PrP d within the LRS. The prevalence of infection was influenced by PRNP genetics at codon 142 and by the age of the goats: methionine carriers older than 60 months showed a much lower prevalence of infection (12/78; 15.4%) than those younger than 60 months (20/42; 47.6%); these last showed prevalence values similar to isoleucine homozygotes of any age (40/80; 50.0%). Two of seven goats with definite signs of scrapie were negative for PrP d in brain but positive in LRS tissues, and one goat showed biochemical and IHC features of PrP d different from all other infected goats. The results of this study have implications for surveillance and control policies for scrapie in goats.scrapie / goat / prion disease / transmissible spongiform encephalopathy

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Section: Implications For Surveillance Of Goat Tsesupporting

confidence: 88%

Section: Implications For Surveillance Of Goat Tsementioning

confidence: 77%

High prevalence of scrapie in a dairy goat herd: tissue distribution of disease-associated PrP and effect ofPRNPgenotype and age

González¹,

Martin²,

Sisó³

et al. 2009

Vet. Res.

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“…Several authors have reported the use of WB to identify the PrP Sc in samples of ovine brain (O'ROURKE et al, 1998;GONZÁLEZ et al, 2006;BILHEUDE et al, 2007). However, for antemortem diagnostics using samples of lymphoid tissues, this method presents a few limitations, owing to the fact that the harvested sample shows a low amount of lymphoid follicles, thus reducing the chance of obtaining detectable amounts of PrP Sc , and increasing the probability of false negative results (LEAL et al, 2012).…”

Section: Western Blottingmentioning

confidence: 99%

“…However, these procedures are not easily performed in the field and do not allow for repeated resampling. The description of constitutive lymphoid tissue in the sheep rectum (ALEKSANDERSEN; NICANDER; LANDSVERK, 1991) prompted the use of rectal mucosa samples for the post-mortem diagnosis of scrapie (GONZÁLEZ et al, 2006), since accumulation of PrP Sc occurs in such tissue. Moreover, samples from the rectal mucosa can also be obtained using a simple and rapid biopsy technique that does not require specific instruments, sedation, or general anesthesia and that can be performed several times without any detrimental effects on the animal.…”

Section: Introductionmentioning

confidence: 99%

Monoclonal and polyclonal antibodies for ante- and post-mortem detection of PrPSc in sheep

Raksa¹,

Costa

González

et al. 2015

Sem. Ci. Agr.

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Scrapie is a disease that affects sheep and goats and is characterized by the accumulation of an abnormal isoform (PrP Sc ) of the cellular prion protein, PrP C , in the central nervous system (CNS) and in lymphoid tissues. Detection of PrP Sc in these tissues can be attempted by a variety of techniques, including immunohistochemistry (IHC) and western blotting (WB), for which a wide range of monoclonal and polyclonal antibodies are commercially available. The objective of this study was to test and compare the efficacy of monoclonal antibodiesF89/160.1.5, F99/97.6.1, and P4 and polyclonal antibodies M52 and R486 in the detection of PrP Sc in lymphoid and CNS tissue samples by using IHC. Positive and negative control samples of sheep brain and tonsils were provided by the Animal Health and Veterinary Laboratories Agency (AHVLA, UK). The IHC examination of CNS samples with both monoclonal and polyclonal antibodies confirmed the granular deposition of PrP Sc in the neurons of the positive control tissues. However, while the monoclonal antibodies did not produce positive reactions in the negative controls, the polyclonal antibodies showed some non-specific staining. The testing of positive control tonsil samples with polyclonal and monoclonal antibodies identified positive control-specific reactions, whereas the negative control tissues were IHC-negative with all antibodies, although P4 and the polyclonal antibodies produced some background staining. In summary, although the polyclonal antibodies may be more accessible, in this study their use is not advisable because of possible false positive reactions. The polyclonal antibody M52 was able to identify PrP C in brain and spleen samples by WB but other lymphoid tissues were negative. ResumoScrapie é uma doença que afeta ovinos e caprinos, sendo definida pelo acúmulo de uma isoforma anormal (PrP Sc ) da proteína priônica celular (PrP C ) no Sistema Nervoso Central (SNC) e em tecidos linfoides. Para as técnicas de eleição do diagnóstico de scrapie, imunohistoquímica (IHQ) e western blotting (WB), podem ser utilizados uma vasta gama de anticorpos monoclonais e policlonais comercialmente disponíveis. O objetivo deste estudo foi testar e comparar a eficácia dos anticorpos monoclonais disponíveis comercialmente, F89/160.1.5, F99/97.6.1 e P4, e os anticorpos policlonais R486 e M52, para a identificação da presença da PrP Sc em amostras de tecido linfoide e SNC através da técnica de IHQ. Foram utilizadas nas avaliações de IHQ amostras positivas e negativas de cérebro e tonsila palatina de ovinos, cedidas pelo Animal Health Veterinary Laboratory Agency (AHVLA), Reino Unido. Para WB, foram utilizadas amostras de encéfalo, baço, linfonodo, terceira pálpebra e mucosa retal de ovino. As análises IHQ utilizando anticorpos monoclonais e policlonais em amostras positivas de cérebro confirmaram a deposição da PrP Sc em neurônios, caracterizada por marcações de aspecto granular intraneural. Na amostra negativa de cérebro, os anticorpos monoclonais não identificaram marcaçõe...

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“…PrP Sc has been found in tissues that could be involved in the natural dissemination of the infectious agent i.e., routes that could lead to exit of the infectious agent from the animal, and result in either environmental contamination or direct transmission. These tissues include the lympho-reticular system of the gut [40,103,115], chronically inflamed mammary tissue associated with lymphocyctic mastitis [73], kidney tissue [90], salivary glands [104], nictitating membrane [77], and placentae [2,81,101].…”

Section: Alternative Disease Modelsmentioning

confidence: 99%

Approaches to investigating transmission of spongiform encephalopathies in domestic animals using BSE as an example

et al. 2008

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-Bovine spongiform encephalopathy was a novel spongiform encephalopathy, in an hitherto unaffected species, that had characteristics of a point source epidemic, with an agent that could have been incorporated into a wide variety of feedstuffs and iatrogenically administered to naïve populations, and there was early evidence that it was not restricted to bovines. It was vital to establish, albeit experimentally, which other species might be affected, and whether the epidemic could be maintained by natural transmission, if the source was removed. In contrast, scrapie has been endemic throughout Great Britain for centuries, is maintained naturally (even if we don't know exactly how) and has a known host range. The principles, process and integration of evidence from different types of studies, however, are similar for both of these transmissible spongiform encephalopathies (TSE) and can be applied to any emerging or suspected spongiform encephalopathy. This review discusses the experimental approaches used to determine TSE transmissibility and infectivity and how they relate to natural disease and control measures.

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Postmortem diagnosis of preclinical and clinical scrapie in sheep by the detection of disease‐associated PrP in their rectal mucosa

Cited by 66 publications

References 30 publications

High prevalence of scrapie in a dairy goat herd: tissue distribution of disease-associated PrP and effect ofPRNPgenotype and age

High prevalence of scrapie in a dairy goat herd: tissue distribution of disease-associated PrP and effect ofPRNPgenotype and age

Monoclonal and polyclonal antibodies for ante- and post-mortem detection of PrPSc in sheep

Approaches to investigating transmission of spongiform encephalopathies in domestic animals using BSE as an example

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