McCutcheon S, Hunter N, Houston F
Institute for Animal Health, Compton, Newbury, Berkshire, RG20 7NN, UK.
J Immunol Methods. 2005 Mar;298(1-2):119-28. doi: 10.1016/j.jim.2005.01.012.
The diagnosis of prion diseases, such as scrapie and BSE, has traditionally relied upon the identification of the disease-associated form of the prion protein, PrP(Sc), based on its resistance to digestion by proteinase K (PK). A more recent development is the conformation-dependent immunoassay (CDI), which distinguishes between PrP Sc and normal PrP (PrP C) based on their differing solubility in guanidine hydrochloride rather than resistance or sensitivity to PK. We have developed a CDI-formatted sandwich immunoassay for the measurement of PrP Sc in sheep brain, which discriminates between clinically affected scrapie cases (natural or experimental) and uninfected controls of the same PrP genotype. Using this method, we have shown for the first time that, in sheep, the PrP genotype has a significant influence on the amount of PrP Sc deposited in the brains of animals experimentally infected with scrapie.
诸如羊瘙痒病和牛海绵状脑病等朊病毒疾病的诊断，传统上依赖于对与疾病相关的朊病毒蛋白形式PrP(Sc)的鉴定，这是基于其对蛋白酶K（PK）消化的抗性。最近的一项进展是构象依赖性免疫测定（CDI），它基于PrP Sc和正常PrP（PrP C）在盐酸胍中不同的溶解性来区分它们，而不是基于对PK的抗性或敏感性。我们开发了一种用于测量羊脑中PrP Sc的CDI格式夹心免疫测定法，该方法能区分临床受影响的瘙痒病病例（自然感染或实验感染）和相同PrP基因型的未感染对照。使用这种方法，我们首次表明，在绵羊中，PrP基因型对实验感染瘙痒病的动物大脑中沉积的PrP Sc量有显著影响。
