α-突触核蛋白实时震颤诱导转化技术在帕金森病中国患者脑和皮肤标本中的应用

Application of α-Syn Real-Time Quaking-Induced Conversion for Brain and Skin Specimens of the Chinese Patients With Parkinson's Disease.

作者信息

Chen Dong-Dong, Jiao Ling, Huang Yue, Xiao Kang, Gao Li-Ping, Chen Cao, Shi Qi, Dong Xiao-Ping

机构信息

State Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Zhejiang University, Beijing, China.

Department of Neurology, The Affiliated Hospital of Guizhou Medical University, Guiyang, China.

出版信息

Front Aging Neurosci. 2022 Jul 1;14:898516. doi: 10.3389/fnagi.2022.898516. eCollection 2022.

DOI:10.3389/fnagi.2022.898516

PMID:35847665

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9283982/

Abstract

The real-time quaking-induced conversion (RT-QuIC) assay has been developed and used as an diagnostic tool for Parkinson's disease (PD). In this study, we established α-Syn RT-QuIC using recombinant human α-Syn as the substrate. All 5 brain homogenates of neuropathological PD cases and 13 skin homogenates of clinical PD cases showed positive results, whereas all the samples of negative controls remain negative. Meantime, randomly selected 6 skin samples of PD cases and 6 skin samples of sCJD cases showed negative in opposite prion RT-QuIC and α-Syn RT-QuIC. Our α-Syn RT-QuIC showed dose-dependent manner between the lag times and peak ThT fluorescent values. Additionally, the detecting limitation was about 10 dilution for brain tissues and 10 for skins. Those data indicate a reliable specificity and good sensitivity of the established α-Syn RT-QuIC in identifying and amplifying the misfolded α-Syn in brain and skin tissues of patients with PD.

摘要

实时震颤诱导转化（RT-QuIC）检测方法已被开发出来，并用作帕金森病（PD）的诊断工具。在本研究中，我们以重组人α-突触核蛋白为底物建立了α-突触核蛋白RT-QuIC检测方法。所有5例神经病理学确诊的PD病例的脑匀浆和13例临床确诊的PD病例的皮肤匀浆均显示阳性结果，而所有阴性对照样本均为阴性。同时，随机选取的6例PD病例的皮肤样本和6例克雅氏病（sCJD）病例的皮肤样本在相反的朊病毒RT-QuIC和α-突触核蛋白RT-QuIC检测中均呈阴性。我们的α-突触核蛋白RT-QuIC在延迟时间和ThT荧光峰值之间呈现剂量依赖性。此外，脑组织的检测限约为10倍稀释，皮肤的检测限约为10倍稀释。这些数据表明，所建立的α-突触核蛋白RT-QuIC在识别和扩增PD患者脑和皮肤组织中错误折叠的α-突触核蛋白方面具有可靠的特异性和良好的敏感性。