Streamlined alpha-synuclein RT-QuIC assay for various biospecimens in Parkinson’s disease and dementia with Lewy bodies

作者：Connor Bargar, Wen Wang, Steven A. Gunzler, Alexandra LeFevre, Zerui Wang, Alan J. Lerner, Neena Singh, Curtis Tatsuoka, Brian S. Appleby, Xiongwei Zhu, Rong Xu, Vahram Haroutunian, Wen‐Quan Zou, Jiyan Ma, Shu G. Chen · 发表于：Acta Neuropathologica Communications · 年份：2021 · DOI：10.1186/s40478-021-01175-w · 被引用次数：221 · 研究领域：Parkinson's Disease Mechanisms and Treatments、Neurological disorders and treatments、Alzheimer's disease research and treatments

Abstract Definitive diagnosis of Parkinson’s disease (PD) and dementia with Lewy bodies (DLB) relies on postmortem finding of disease-associated alpha-synuclein (αSyn D ) as misfolded protein aggregates in the central nervous system (CNS). The recent development of the real-time quaking induced conversion (RT-QuIC) assay for ultrasensitive detection of αSyn D aggregates has revitalized the diagnostic values of clinically accessible biospecimens, including cerebrospinal fluid (CSF) and peripheral tissues. However, the current αSyn RT-QuIC assay platforms vary widely and are thus challenging to implement and standardize the measurements of αSyn D across a wide range of biospecimens and in different laboratories. We have streamlined αSyn RT-QuIC assay based on a second generation assay platform that was assembled entirely with commercial reagents. The streamlined RT-QuIC method consisted of a simplified protocol requiring minimal hands-on time, and allowing for a uniform analysis of αSyn D in different types of biospecimens from PD and DLB. Ultrasensitive and specific RT-QuIC detection of αSyn D aggregates was achieved in million-fold diluted brain homogenates and in nanoliters of CSF from PD and DLB cases but not from controls. Comparative analysis revealed higher seeding activity of αSyn D in DLB than PD in both brain homogenates and CSF. Our assay was further validated with CSF samples of 214 neuropathologically confirmed cases from tissue repositories (88 PD, 58 DLB, and 68 ...