Early detection of abnormal prion protein in genetic human prion diseases now possible using real-time QUIC assay.
Early detection of abnormal prion protein in genetic human prion diseases now possible using real-time QUIC assay.
复制标题
DOI：
10.1371/journal.pone.0054915
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Nishida N
中科院分区：
文献类型：
--
作者：
Sano K;Satoh K;Atarashi R;Takashima H;Iwasaki Y;Yoshida M;Sanjo N;Murai H;Mizusawa H;Schmitz M;Zerr I;Kim YS;Nishida N
遗传性朊病毒病（gPrD）的明确诊断需要病理学证实。迄今为止，诊断依赖于在脑脊液（CSF）中发现生物标志物14-3-3蛋白和总tau（t-tau）蛋白，但许多研究人员报告说，这些标志物在gPrD中没有充分升高，特别是在Gerstmann-Sträussler-Scheinker综合征（GSS）中。我们最近开发了一种新的体外扩增技术，命名为“实时quaking-induced转换（RT-QUIC）”，以检测从散发性克雅氏病（sCJD）患者的CSF中的朊蛋白的异常形式。在本研究中，我们的目的是研究生物标志物的存在，并评估RT-QUIC测定在gPrD患者中的作用，因为RT-QUIC作为gPrD诊断工具的效用尚未确定。从gPrD患者中获得56份CSF样本，包括20例P102 L突变的GSS，12例致死性家族性失眠症（FFI; D178 N）和24例遗传性CJD（gCJD），其中22例E200 K突变和2例V203 I突变。我们将所有CSF样品进行RT-QUIC测定，通过Western印迹分析14-3-3蛋白，并使用ELISA试剂盒测量t-tau蛋白。RT-QUIC的检测灵敏度如下：GSS（78%）、FFI（100%）、gCJD E200 K（87%）和gCJD V203 I（100%）。另一方面，生物标志物的检测灵敏度相当低：GSS（11%），FFI（0%），gCJD E200 K（73%）和gCJD V203 I（67%）。因此，与生物标志物检测相比，RT-QUIC具有更高的检测灵敏度，特别是在GSS和FFI患者中。RT-QUIC测定比检测gPrD患者的生物标志物更敏感。因此，当患者或患者家属不同意进行基因检测时，RT-QUIC方法可用作诊断工具，或者在基因检测结果呈阳性的情况下确认诊断。
The definitive diagnosis of genetic prion diseases (gPrD) requires pathological confirmation. To date, diagnosis has relied upon the finding of the biomarkers 14-3-3 protein and total tau (t-tau) protein in the cerebrospinal fluid (CSF), but many researchers have reported that these markers are not sufficiently elevated in gPrD, especially in Gerstmann-Sträussler-Scheinker syndrome (GSS). We recently developed a new in vitro amplification technology, designated “real-time quaking-induced conversion (RT-QUIC)”, to detect the abnormal form of prion protein in CSF from sporadic Creutzfeldt-Jakob disease (sCJD) patients. In the present study, we aimed to investigate the presence of biomarkers and evaluate RT-QUIC assay in patients with gPrD, as the utility of RT-QUIC as a diagnostic tool in gPrD has yet to be determined. 56 CSF samples were obtained from gPrD patients, including 20 cases of GSS with P102L mutation, 12 cases of fatal familial insomnia (FFI; D178N), and 24 cases of genetic CJD (gCJD), comprising 22 cases with E200K mutation and 2 with V203I mutation. We subjected all CSF samples to RT-QUIC assay, analyzed 14-3-3 protein by Western blotting, and measured t-tau protein using an ELISA kit. The detection sensitivities of RT-QUIC were as follows: GSS (78%), FFI (100%), gCJD E200K (87%), and gCJD V203I (100%). On the other hand the detection sensitivities of biomarkers were considerably lower: GSS (11%), FFI (0%), gCJD E200K (73%), and gCJD V203I (67%). Thus, RT-QUIC had a much higher detection sensitivity compared with testing for biomarkers, especially in patients with GSS and FFI. RT-QUIC assay is more sensitive than testing for biomarkers in gPrD patients. RT-QUIC method would thus be useful as a diagnostic tool when the patient or the patient's family does not agree to genetic testing, or to confirm the diagnosis in the presence of a positive result for genetic testing.
登录
查看更多内容
DOI：
10.1097/00005072-199810000-00010
发表时间：
1998-10-01
影响因子：
3.2
作者：
Piccardo, P;Dlouhy, SR;Ghetti, B
通讯作者：
Ghetti, B
p102l遗传性prion病的表型异质性和国际遗传疾病的遗传修饰。
影响因子：
14.5
作者：
Webb, T. E. F.;Poulter, M.;Beck, J.;Uphill, J.;Adamson, G.;Campbell, T.;Linehan, J.;Powell, C.;Brandner, S.;Pal, S.;Siddique, D.;Wadsworth, J. D.;Joiner, S.;Alner, K.;Petersen, C.;Hampson, S.;Rhymes, C.;Treacy, C.;Storey, E.;Geschwind, M. D.;Nemeth, A. H.;Wroe, S.;Collinge, J.;Mead, S.
通讯作者：
Mead, S.
影响因子：
3.5
作者：
Popova, S. N.;Tarvainen, I.;Alafuzoff, I.
通讯作者：
Alafuzoff, I.
影响因子：
82.9
作者：
Atarashi, Ryuichiro;Satoh, Katsuya;Nishida, Noriyuki
通讯作者：
Nishida, Noriyuki
影响因子：
5
作者：
Satoh, Katsuya;Tobiume, Minoru;Sata, Testutaro
通讯作者：
Sata, Testutaro