A new model for sensitive detection of zoonotic prions by PrP transgenic Drosophila.
A new model for sensitive detection of zoonotic prions by PrP transgenic Drosophila.
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DOI：
10.1016/j.jbc.2021.100878
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发表时间：
2021-08
期刊：
影响因子：
--
通讯作者：
Bujdoso R
中科院分区：
文献类型：
--
作者：
Thackray AM;Andréoletti O;Spiropoulos J;Bujdoso R
朊病毒是一种可传播的蛋白质病原体，在合适的宿主（通常是小鼠）中通过生物测定法最可靠地检测到。然而，小鼠生物测定是缓慢和繁琐的，并且对低滴度的朊病毒感染性相对不敏感。朊病毒可以通过蛋白质错误折叠循环扩增（PMCA）技术在体外进行生物化学检测，该技术扩增疾病相关朊病毒蛋白，但不能检测真正的朊病毒感染性。在这里，我们证明，果蝇转基因牛朊病毒蛋白（PrP）的表达可以作为一个模型系统，用于检测牛朊病毒显着更有效地比小鼠朊病毒生物测定或PMCA。引人注目的是，牛PrP转基因果蝇可以在10 - 12稀释的经典牛海绵状脑病（BSE）接种物中检测到牛朊病毒感染性，这比牛PrP小鼠生物测定法灵敏106倍。牛PrP转基因果蝇检测H型和L型非典型BSE和绵羊传代BSE的敏感性相似。果蝇中牛朊病毒的生物测定在7周内进行，而小鼠朊病毒生物测定需要至少一年来评估相同的接种物。此外，牛PrP转基因果蝇可以检测到经典BSE的水平比PMCA达到的水平低105倍。这些数据表明，PrP转基因果蝇代表了一种新的易于处理的朊病毒生物测定法，用于有效和灵敏地检测哺乳动物朊病毒，包括那些已知的人畜共患病的潜力。
Prions are transmissible protein pathogens most reliably detected by a bioassay in a suitable host, typically mice. However, the mouse bioassay is slow and cumbersome, and relatively insensitive to low titers of prion infectivity. Prions can be detected biochemically in vitro by the protein misfolding cyclic amplification (PMCA) technique, which amplifies disease-associated prion protein but does not detect bona fide prion infectivity. Here, we demonstrate that Drosophila transgenic for bovine prion protein (PrP) expression can serve as a model system for the detection of bovine prions significantly more efficiently than either the mouse prion bioassay or PMCA. Strikingly, bovine PrP transgenic Drosophila could detect bovine prion infectivity in the region of a 10−12 dilution of classical bovine spongiform encephalopathy (BSE) inoculum, which is 106-fold more sensitive than that achieved by the bovine PrP mouse bioassay. A similar level of sensitivity was observed in the detection of H-type and L-type atypical BSE and sheep-passaged BSE by bovine PrP transgenic Drosophila. Bioassays of bovine prions in Drosophila were performed within 7 weeks, whereas the mouse prion bioassay required at least a year to assess the same inoculum. In addition, bovine PrP transgenic Drosophila could detect classical BSE at a level 105-fold lower than that achieved by PMCA. These data show that PrP transgenic Drosophila represent a new tractable prion bioassay for the efficient and sensitive detection of mammalian prions, including those of known zoonotic potential.
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影响因子：
4.4
作者：
Benestad SL;Mitchell G;Simmons M;Ytrehus B;Vikøren T
通讯作者：
Vikøren T
DOI：
10.1073/pnas.0305777101
发表时间：
2004-03-02
影响因子：
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影响因子：
2.7
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