Dong C-F, Huang Y-X, An R, Chen J-M, Wang X-F, Shan B, Lei Y-J, Han L, Zhang B-Y, Han J, Dong X-P
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Prevention and Control, Chinese Center for Disease Control and Prevention, Ying-Xin Rd 100, Beijing 100052, China.
Zoonoses Public Health. 2007;54(8):328-36. doi: 10.1111/j.1863-2378.2007.01062.x.
Transmissible spongiform encephalopathies, also termed prion diseases, are fatal neurodegenerative disorders that affect both humans and animals, which are characterized by presences of protease-resistance disease-associated prion protein (PrP(Sc)) in brains. In the present study, we optimized the Western blot assay for PrP(Sc) with a precipitation procedure of streptomycin sulphate. After incubated with suitable amount of streptomycin sulphate, the detective sensitivity for PrP(Sc) was remarkably improved. The precipitation of PrP(Sc) was obviously influenced by pH value in the solution. Employs of PrP(Sc) stock sample into various mimic specimens, including normal hamster brain homogenate, human cerebrospinal fluid and urine, demonstrated that streptomycin precipitation markedly increased the detective sensitivity of PrP(Sc), regardless in low concentration or in large volume. In addition, the PrP(Sc) from a human brain tissue of familiar Creutzfeldt-Jakob disease (fCJD) was efficiently precipitated with streptomycin sulphate. As a sensitive, specific, rapid and flexible protocol for PrP(Sc), the protocol in this study has the potential, alone or combined with other techniques, to detect low levels of PrP(Sc) in the specimens not only from central nerve system, but also from peripheral organs or fluids.
传染性海绵状脑病，也称为朊病毒病，是一种致命的神经退行性疾病，可影响人类和动物，其特征是大脑中存在抗蛋白酶的疾病相关朊病毒蛋白（PrP(Sc)）。在本研究中，我们通过硫酸链霉素沉淀法优化了PrP(Sc)的蛋白质印迹检测方法。在与适量硫酸链霉素孵育后，对PrP(Sc)的检测灵敏度显著提高。PrP(Sc)的沉淀明显受溶液pH值的影响。将PrP(Sc)储备样品应用于各种模拟标本，包括正常仓鼠脑匀浆、人脑脊液和尿液，结果表明，无论浓度低还是体积大，硫酸链霉素沉淀均显著提高了PrP(Sc)的检测灵敏度。此外，来自家族性克雅氏病（fCJD）人脑组织的PrP(Sc)能被硫酸链霉素有效沉淀。作为一种针对PrP(Sc)的灵敏、特异、快速且灵活的方法，本研究中的方法单独或与其他技术结合使用，有潜力检测不仅来自中枢神经系统，还来自外周器官或体液的标本中的低水平PrP(Sc)。
