Masujin Kentaro, Orrú Christina D, Miyazawa Kohtaro, Groveman Bradley R, Raymond Lynne D, Hughson Andrew G, Caughey Byron
Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA Influenza and Prion Disease Research Center, National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Japan.
Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
J Clin Microbiol. 2016 Mar;54(3):676-86. doi: 10.1128/JCM.02731-15. Epub 2016 Jan 6.
Prion diseases of cattle include the classical bovine spongiform encephalopathy (C-BSE) and the atypical H-type BSE (H-BSE) and L-type BSE (L-BSE) strains. Although the C- and L-BSE strains can be detected and discriminated by ultrasensitive real-time quaking-induced conversion (RT-QuIC) assays, no such test has yet been described for the detection of H-BSE or the discrimination of each of the major bovine prion strains. Here, we demonstrate an RT-QuIC assay for H-BSE that can detect as little as 10(-9) dilutions of brain tissue and neat cerebrospinal fluid samples from clinically affected cattle. Moreover, comparisons of the reactivities with different recombinant prion protein substrates and/or immunoblot band profiles of proteinase K-treated RT-QuIC reaction products indicated that H-, L-, and C-BSE have distinctive prion seeding activities and can be discriminated by RT-QuIC on this basis.
牛的朊病毒病包括经典牛海绵状脑病（C-BSE）以及非典型H型BSE（H-BSE）和L型BSE（L-BSE）毒株。尽管C型和L型BSE毒株可通过超灵敏实时震颤诱导转化（RT-QuIC）检测法进行检测和区分，但尚未有用于检测H-BSE或区分各主要牛朊病毒毒株的此类检测方法。在此，我们展示了一种针对H-BSE的RT-QuIC检测法，该方法能够检测出临床患病牛脑组织和纯脑脊液样本低至10⁻⁹稀释度的样本。此外，对不同重组朊病毒蛋白底物的反应性以及蛋白酶K处理后的RT-QuIC反应产物的免疫印迹条带图谱进行比较表明，H型、L型和C型BSE具有独特的朊病毒播种活性，并可据此通过RT-QuIC进行区分。
