Discrimination between L-type and C-type bovine spongiform encephalopathy by the strain-specific reactions of real-time quaking-induced conversion
Discrimination between L-type and C-type bovine spongiform encephalopathy by the strain-specific reactions of real-time quaking-induced conversion
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通过实时震动诱导转换的菌株特异性反应区分 L 型和 C 型牛海绵状脑病
DOI：
10.1016/j.bbrc.2020.03.183
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发表时间：
2020
影响因子：
3.1
通讯作者：
Atarashi Ryuichiro
中科院分区：
文献类型：
--
作者：
Ubagai Kaori;Fukuda Shigeo;Mori Tsuyoshi;Takatsuki Hanae;Taguchi Yuzuru;Kageyama Soichi;Nishida Noriyuki;Atarashi Ryuichiro
实时振荡诱导转换（RT-QUIC）检测使用大肠杆菌衍生的纯化重组朊蛋白（rPrP），使我们能够从标本中扩增痕量的异常形式的PrP（PrPSc）。该技术可用于人类和动物朊病毒疾病的早期诊断和朊病毒污染的评估。在本研究中，我们证明了非典型形式的牛海绵状脑病（BSE），l-BSE和经典BSE（C-BSE）之间的RT-QUIC反应存在菌株特异性差异。小鼠rPrP（rMoPrP）在来自I-BSE或C-BSE的PrPScseed存在下有效地转化为淀粉样纤维，而仓鼠rPrP（rHaPrP）仅在I-BSE中转化，而不是C-BSE。这些特征在第二轮反应中得以保留，但在随后的几轮中逐渐减弱，到第五轮时完全丧失，这很可能是由于RT-QUIC中非特异性rPrP淀粉样蛋白原纤维的选择性生长优势。我们的发现进一步增强了使用RT-QUIC对朊病毒菌株的区分，并进一步加深了我们对朊病毒菌株分子基础的理解。
Real-time quaking-induced conversion (RT-QUIC) assays usingEscherichia coli-derived purified recombinant prion protein (rPrP) enable us to amplify a trace amount of the abnormal form of PrP (PrPSc) from specimens. This technique can be useful for the early diagnosis of both human and animal prion diseases and the assessment of prion contamination. In the present study, we demonstrated that there are strain-specific differences in the RT-QUIC reactions between an atypical form of bovine spongiform encephalopathy (BSE),l-BSE, and classical BSE (C-BSE). Whereas mouse rPrP (rMoPrP) was efficiently converted to amyloid fibrils in the presence of PrPScseed derived from eitherl-BSE or C-BSE, hamster rPrP (rHaPrP) was converted only inl-BSE, not C-BSE. These characteristics were preserved in the second round reaction, but gradually weakened in the subsequent rounds and were completely lost by the fifth round, most likely due to the selective growth advantage of nonspecific rPrP amyloid fibrils in the RT-QUIC. Our findings further enhance the discrimination of prion strains using RT-QUIC, and further our understanding of the molecular basis of prion strains.