Demographic characteristics of scrapie-affected holdings identified by active and passive surveillance schemes in Great Britain: 2002–2005. PrP genotype: A flock-level risk factor for scrapie? Preventive Veterinary Medicine, 92, 309–323. Preventive Veterinary Medicine, 109, 179–184. BSE and variant CJD: Emerging science, public pressure and the vagaries of policy-making BSE and variant CJD: Emerging science, public pressure and the vagaries of policy-making. Preventive Veterinary Medicine, 105, 255–264. Geographical BSE risk assessment and its impact on disease detection and dissemination. Salman, M., Silano, V., Heim, D., & Kreysa, J. Journal of Molecular Biology, 412, 660–673. Toward the molecular basis of inherited prion diseases: NMR structure of the human prion protein with V210I mutation. The aim of this study was to develop a duplex real-time PCR assay followed by a melt curve step for sensitive, rapid, specific, and cost-effective detection of bovine and caprine species based on the amplicon melting peak in ruminant feeds to prevent Transmissible Spongiform Encephalopathies.īiljan, I., llc, G., Giachin, G., Raspadori, A., Zhukov, I., Plavec, J., et al. Duplex analysis of the reference samples showed that the detection limit of the assay was 0.003 % for bovine and 0.005 % for caprine species. Gene products of caprine and bovine produced two distinct melting peaks simultaneously at 82 and 86.8 ☌, respectively. DNA was isolated from heat-treated meats (133 ☌/3 bar for 20 min) mixtures of bovine and caprine and was used to optimize the assay. The method merges the use of bovine ( Bos taurus) and caprine ( Capra hircus) specific primers that amplify small fragments (bovine 96 bp and caprine 142 bp) of the mitochondrial 16S rRNA and 12S rRNA genes, respectively. A duplex real-time PCR assay with melting curve analysis, using the EvaGreen fluorescence dye, was developed for rapid and reliable identification of bovine and caprine in ruminant feeds.
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