Comparison of Immunohistochemistry and Two Rapid Tests for Detection of Abnormal Prion Protein in Different Brain Regions of Sheep with Typical Scrapie

Bolea, Rosa; Monleón, Eva; Schiller, Irene; Raeber, A.; Acín, Cristina; Monzón, Marta; Martín-Burriel, Inmaculada; Struckmeyer, Thomas; Oesch, Bruno; Badiola, Juan José

doi:10.1177/104063870501700511

Cited by 28 publications

(25 citation statements)

References 11 publications

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“…PRNP genotypes were determined according to Acin et al (2004). The diagnosis was confirmed using a rapid test and immunohistochemistry to detect PrP Sc (Bolea et al 2005). These animals belong to a flock of sheep, conserved for research purposes by the Prion Research Centre of the University of Zaragoza and where several SC cases have appeared in the last 4 years.…”

Section: Chromosomal Locationmentioning

confidence: 99%

“…All these animals belong to six flocks of sheep, conserved for research purposes by the Prion Research Centre of the University of Zaragoza and where several SC cases have appeared in the last 4 years. PRNP genotypes and SC diagnosis were assessed as explained before (Acin et al 2004;Bolea et al 2005). To determine if the SNP's genotype distribution found in this group of animals was representative of other breeds or exclusive of the Rasa Aragonesa group, an additional genotyping of 38 ARQ/ ARQ sheep belonging to the Manchega and Assaf breeds, which had no contact with SC, was carried out.…”

Section: Polymorphism Detectionmentioning

confidence: 99%

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Structural and functional analysis of the HSP90AA1 gene: distribution of polymorphisms among sheep with different responses to scrapie

Marcos-Carcavilla¹,

Calvo²,

González³

et al. 2005

Cell Stress Chaper

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Scrapie is a transmissible spongiform encephalopathy in sheep and goats. Susceptibility to this neurodegenerative disease is mainly controlled by point mutations at the PRNP locus. Other genes, apart from PRNP, have been reported to modulate resistance/susceptibility to scrapie. On the basis of several studies in Alzheimer and different transmissible spongiform encephalopathy models, HSP90AA1 was chosen as a putative positional and functional candidate gene that might be involved in the polygenic variance mentioned above. In the present work, the ovine HSP90AA1 gene including the promoter and other regulatory regions has been isolated and characterized. Several sequence polymorphisms have also been identified. FISH-mapping localized the HSP90AA1 gene on ovine chromosome OAR19q24dist, which was confirmed by linkage analysis. This chromosome region has been shown to include a quantitative trait loci (QTL) for scrapie incubation period in sheep. Expression analyses were carried out in spleen and cerebellum samples. No differences in the expression of the HSP90AA1 gene were found in any of these tissues (p>0.05) between control and infected animal samples. Nevertheless, association analyses revealed that several polymorphisms in the 5′ and 3′ regions of the HSP90AA1 gene were differentially distributed among animals with different responses to scrapie infection. Thus, results presented here support the hypothesis that HSP90AA1 could be a positional and functional candidate gene modulating the response to scrapie in sheep.

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Section: Chromosomal Locationmentioning

confidence: 99%

Section: Polymorphism Detectionmentioning

confidence: 99%

Structural and functional analysis of the HSP90AA1 gene: distribution of polymorphisms among sheep with different responses to scrapie

Marcos-Carcavilla¹,

Calvo²,

González³

et al. 2005

Cell Stress Chaper

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“…Res (Bolea et al, 2005). More recently, in vitro amplification methods [serial protein misfolding cyclic amplification (sPMCA) (Saborio et al, 2001), amyloid seeding assay (ASA) (Colby et al, 2007) and real-time quaking-induced conversion (RT-QuIC) (Atarashi et al, 2011a)], all of which employ conversion of substrate PrP C to the disease-specific conformation, have expanded the sensitivity and mechanistic possibilities of prion detection assays.…”

Section: Detection Of Prpmentioning

confidence: 99%

Quantitative assessment of prion infectivity in tissues and body fluids by real-time quaking-induced conversion

Henderson

Davenport

Haley

et al. 2015

Journal of General Virology

103

134

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Prions are amyloid-forming proteins that cause transmissible spongiform encephalopathies through a process involving the templated conversion of the normal cellular prion protein (PrP C ) to a pathogenic misfolded conformation. Templated conversion has been modelled in several in vitro assays, including serial protein misfolding amplification, amyloid seeding and real-time quakinginduced conversion (RT-QuIC). As RT-QuIC measures formation of amyloid fibrils in real-time, it can be used to estimate the rate of seeded conversion. Here, we used samples from deer infected with chronic wasting disease (CWD) in RT-QuIC to show that serial dilution of prion seed was linearly related to the rate of amyloid formation over a range of 10 "3 to 10 "8 mg. We then used an amyloid formation rate standard curve derived from a bioassayed reference sample (CWD+ brain homogenate) to estimate the prion seed concentration and infectivity in tissues, body fluids and excreta. Using these methods, we estimated that urine and saliva from CWD-infected deer both contained 1-5 LD 50 per 10 ml. Thus, over the 1-2 year course of an infection, a substantial environmental reservoir of CWD prion contamination accumulates.

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“…The absence of falsepositive results and the identification of 2 BSE cases in poor-quality samples confirm the excellent performance of WB on samples from fallen stock, as previously reported. 4,6,14 On the other hand, ELISA and ICA exhibited false-positive rates as high as 84.30 and 8.48 per 100,000 negative samples tested, respectively. The ELISA on fallen stock gave an overall false-positive rate statistically higher than both WB and ICA and also about 90 times higher than the rate observed on slaughterhouse samples.…”

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confidence: 99%

Evaluation of Three Rapid Diagnostic Tests Used in Bovine Spongiform Encephalopathy Monitoring in Italy

et al. 2009

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Abstract. In 2001, a compulsory active surveillance system was started in the European Union to assess the prevalence of bovine spongiform encephalopathy (BSE) in the cattle population. The aim of the current study was to report on the field performances of 3 rapid tests: a Western blot (WB), a chemiluminescence enzymelinked immunosorbent assay (ELISA), and an immunochromatographic assay, routinely used at 3 laboratories of the Istituto Zooprofilattico Sperimentale of Lombardia and Emilia Romagna, over 8 years of BSE monitoring activity. A total of 2,802,866 samples from slaughtered animals and 202,453 samples from fallen stock were tested by 1 of 3 tests. Positive results of the rapid tests were confirmed by histopathological examination, immunohistochemistry, and confirmatory WB. The field performances (i.e., initial reactive and false-positive rates) and practical aspects regarding resources and applicability of the tests to high-throughput routine testing laboratories were evaluated. The 3 tests proved to be reliable tools when applied to slaughtered samples, showing no or very low false-positive rates (,1 per 100,000 negative samples tested) and low retesting frequencies (0.02-0.26%). When samples from fallen stock were analyzed, performances of the immunochromatographic assay, and especially the chemiluminescence ELISA, were negatively affected, resulting in higher false-positive and retesting rates. On the other hand, both tests are less expensive, much easier to use, provide more rapid results, and adapt well to application in routine laboratories as compared with WB. In the authors' experience, the immunochromatographic assay was a good compromise between performance and convenience.

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Comparison of Immunohistochemistry and Two Rapid Tests for Detection of Abnormal Prion Protein in Different Brain Regions of Sheep with Typical Scrapie

Cited by 28 publications

References 11 publications

Structural and functional analysis of the HSP90AA1 gene: distribution of polymorphisms among sheep with different responses to scrapie

Structural and functional analysis of the HSP90AA1 gene: distribution of polymorphisms among sheep with different responses to scrapie

Quantitative assessment of prion infectivity in tissues and body fluids by real-time quaking-induced conversion

Evaluation of Three Rapid Diagnostic Tests Used in Bovine Spongiform Encephalopathy Monitoring in Italy

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