Detection of Prions

朊病毒蛋白的病理亚型（PrPres）可以作为朊病毒疾病的标志物，但需要更多的实际测试来进行临床前诊断和敏感检测许多朊病毒感染。之前我们已经证明，震荡诱导转化（QuIC）实验可以检测瘙痒病感染仓鼠脑组织中PrPres的亚飞图水平，并区分正常和瘙痒病感染仓鼠的脑脊液（CSF）样本。在2009财年，我们对具有医学和农业意义的朊病毒疾病：人类变异型克雅氏病（vCJD）和羊痒病进行了快速反应。人类和绵羊脑匀浆样品的朊病毒阳性和阴性在1-2天内进行区分，灵敏度为10-100飞图PrPres。更重要的是，在短短32小时内，我们就能区分出羊瘙痒病感染和未感染羊的脑脊液样本。这是首次在痒病阳性衍生的脑脊液中检测到PrPres。这种新方法能够相对快速和灵敏地检测人CJD和羊痒病PrPres，并可能促进实用的临床前诊断试验的发展。

The pathological isoform of the prion protein (PrPres) can serve as a marker for prion diseases, but more practical tests are needed for preclinical diagnosis and sensitive detection of many prion infections. Previously we showed that the quaking-induced conversion (QuIC) assay can detect sub-femtogram levels of PrPres in scrapie-infected hamster brain tissue and distinguish cerebral spinal fluid (CSF) samples from normal and scrapie-infected hamsters. During FY2009, we adapted the QuIC reaction to prion diseases of medical and agricultural interest: human variant Creutzfeldt-Jakob disease (vCJD) and sheep scrapie. Prion-positive and negative brain homogenate samples from humans and sheep were discriminated within 1-2 days with a sensitivity of 10-100 femtograms PrPres. More importantly, in as little as 32h we were able to distinguish CSF samples from scrapie-infected and uninfected sheep. This is the first time that PrPres is detected in scrapie positive derived CSF. This new method enables the relatively rapid and sensitive detection of human CJD and sheep scrapie PrPres and may facilitate the development of practical preclinical diagnostic tests.