Single-Molecule Counting Coupled to Rapid Amplification Enables Detection of α-Synuclein Aggregates in Cerebrospinal Fluid of Parkinson's Disease Patients.
Single-Molecule Counting Coupled to Rapid Amplification Enables Detection of α-Synuclein Aggregates in Cerebrospinal Fluid of Parkinson's Disease Patients.
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DOI：
10.1002/anie.202014898
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发表时间：
2021-05-17
期刊：
影响因子：
--
通讯作者：
Sierecki E
中科院分区：
文献类型：
--
作者：
Bhumkar A;Magnan C;Lau D;Jun ESW;Dzamko N;Gambin Y;Sierecki E
α-突触核蛋白聚集是帕金森病的标志，也是早期检测和评估疾病进展的有前景的生物标志物。帕金森病分子检测的前景随着基于突触核蛋白种子扩增的检测方法（例如RT‐QuIC或PMCA）的最新发展而实现。在这里，我们采用单分子计数方法，使用简单的3D打印显微镜检测脑脊液（CSF）中的α-突触核蛋白聚集体。单分子方法能够探测RT-QuIC中使用的扩增过程中的早期事件，并精确计数ThT阳性聚集体。重要的是，使用单分子计数还可以对样本进行精细表征，并对患者CSF中存在的蛋白质聚集体进行指纹分析。与对照组相比，个体聚集体的大小和反应性的指纹图谱显示了每个PD患者的独特特征，并可能在未来提供对突触核蛋白病的新见解。建立了一种新的单分子方法用于检测α-突触核蛋白纤维，这是帕金森病（PD）的一种有前途的生物标志物。与直接检测相比，对单个ThT（+）聚集体进行指纹分析和扩增以将检测限提高3个数量级。检测到PD和对照样品之间的行为差异。
α‐Synuclein aggregation is a hallmark of Parkinson's disease and a promising biomarker for early detection and assessment of disease progression. The prospect of a molecular test for Parkinson's disease is materializing with the recent developments of detection methods based on amplification of synuclein seeds (e.g. RT‐QuIC or PMCA). Here we adapted single‐molecule counting methods for the detection of α‐synuclein aggregates in cerebrospinal fluid (CSF), using a simple 3D printed microscope. Single‐molecule methods enable to probe the early events in the amplification process used in RT‐QuIC and a precise counting of ThT‐positive aggregates. Importantly, the use of single‐molecule counting also allows a refined characterization of the samples and fingerprinting of the protein aggregates present in CSF of patients. The fingerprinting of size and reactivity of individual aggregate shows a unique signature for each PD patients compared to controls and may provide new insights on synucleinopathies in the future. A new single‐molecule method was established for the detection of α‐synuclein fibrils, a promising biomarker of Parkinson's disease (PD). Individual ThT(+) aggregates were fingerprinted and amplified to improve detection limits by 3 orders of magnitude, compared to direct detection. A difference of behaviour was detected between PD and control samples.
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影响因子：
7.4
作者：
Kumar, Manjeet;Hong, Yuning;Carver, John A.
通讯作者：
Carver, John A.
DOI：
10.1016/j.bbapap.2010.04.001
发表时间：
2010-07
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Biancalana M;Koide S
通讯作者：
Koide S
影响因子：
168.9
作者：
Chartier-Harlin, MC;Kachergus, J;Destée, A
通讯作者：
Destée, A
影响因子：
3.4
作者：
Lesage, Suzanne;Houot, Marion;Brice, Alexis
通讯作者：
Brice, Alexis
影响因子：
8.6
作者：
Kang, Un Jung;Boehme, Amelia K.;Soto, Claudio
通讯作者：
Soto, Claudio