Detection of prions in blood
Detection of prions in blood
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DOI：
10.1038/nm1286
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发表时间：
2005-09-01
期刊：
影响因子：
82.9
通讯作者：
Soto, C
中科院分区：
文献类型：
--
作者：
Castilla, J;Saá, P;Soto, C
朊病毒疾病是由一种被称为朊病毒的非常规传染因子引起的，主要由错误折叠的朊病毒蛋白（PrPSc）组成(1)。开发高灵敏度的血液中PrPSc生化检测方法是减少疾病传播的首要任务(2)。本研究表明，蛋白质错误折叠循环扩增（PMCA）技术(3)可以自动化和优化，用于PrPSc的高效扩增。我们表明140个PMCA循环导致灵敏度比标准检测方法增加6600倍。连续两轮的PMCA循环使灵敏度提高了1000万倍，并且能够检测到8000个等效的PrPSc分子。值得注意的是，序列PMCA能够以89%的灵敏度和100%的特异性检测痒病仓鼠血样中的PrPSc。这些发现代表了PrPSc首次在血液中被生化检测到，为开发一种非侵入性的方法来早期诊断朊病毒疾病提供了希望。
Prion diseases are caused by an unconventional infectious agent termed prion, composed mainly of the misfolded prion protein ( PrPSc)(1). The development of highly sensitive assays for biochemical detection of PrPSc in blood is a top priority for minimizing the spread of the disease(2). Here we show that the protein misfolding cyclic amplification ( PMCA) technology(3) can be automated and optimized for high- efficiency amplification of PrPSc. We show that 140 PMCA cycles leads to a 6,600- fold increase in sensitivity over standard detection methods. Two successive rounds of PMCA cycles resulted in a 10 million - fold increase in sensitivity and a capability to detect as little as 8,000 equivalent molecules of PrPSc. Notably, serial PMCA enables detection of PrPSc in blood samples of scrapie- afflicted hamsters with 89% sensitivity and 100% specificity. These findings represent the first time that PrPSc has been detected biochemically in blood, offering promise for developing a noninvasive method for early diagnosis of prion diseases.