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A Simple Versatile and Sensitive Cell-Based Assay for Prions from Various Species

机译:一种简单通用和灵敏的基于细胞的各种物种Pr病毒检测方法

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摘要

Detection and quantification of prion infectivity is a crucial step for various fundamental and applied aspects of prion research. Identification of cell lines highly sensitive to prion infection led to the development of cell-based titration procedures aiming at replacing animal bioassays, usually performed in mice or hamsters. However, most of these cell lines are only permissive to mouse-adapted prions strains and do not allow titration of prions from other species. In this study, we show that epithelial RK13, a cell line permissive to mouse and bank vole prion strains and to natural prion agents from sheep and cervids, enables a robust and sensitive detection of mouse and ovine-derived prions. Importantly, the cell culture work is strongly reduced as the RK13 cell assay procedure designed here does not require subcultivation of the inoculated cultures. We also show that prions effectively bind to culture plastic vessel and are quantitatively detected by the cell assay. The possibility to easily quantify a wider range of prions, including rodent experimental strains but also natural agents from sheep and cervids, should prompt the spread of cell assays for routine prion titration and lead to valuable information in fundamental and applied studies.
机译:对and病毒感染性的检测和定量是of病毒研究的各个基础和应用方面的关键步骤。对to病毒感染高度敏感的细胞系的鉴定导致了基于细胞的滴定程序的发展,该程序旨在替代通常在小鼠或仓鼠中进行的动物生物测定。然而,大多数这些细胞系仅允许小鼠适应的mouse病毒株,并且不能滴定其他物种的病毒。在这项研究中,我们表明,上皮细胞RK13(一种允许小鼠和岸田病毒毒株和羊和宫颈天然natural病毒制剂允许的细胞系)能够对小鼠和绵羊来源的病毒进行稳健而灵敏的检测。重要的是,细胞培养工作大大减少,因为此处设计的RK13细胞分析程序不需要对培养的培养物进行继代培养。我们还显示病毒有效结合到培养塑料容器,并通过细胞测定法进行定量检测。容易量化更广泛范围的of病毒的可能性,包括啮齿动物实验株,还有绵羊和宫颈的天然制剂,应促使常规assay病毒滴定的细胞测定法普及,并在基础和应用研究中获得有价值的信息。

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