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脑脊液中α-突触核蛋白检测新方法的验证

Validation of a new assay for α-synuclein detection in cerebrospinal fluid.

作者信息

Førland Marthe Gurine, Öhrfelt Annika, Oftedal Linn Silje, Tysnes Ole-Bjørn, Larsen Jan Petter, Blennow Kaj, Zetterberg Henrik, Alves Guido, Lange Johannes

出版信息

Clin Chem Lab Med. 2017 Feb 1;55(2):254-260. doi: 10.1515/cclm-2016-0409.

Abstract

BACKGROUND

Abnormal α-synuclein aggregation and deposition is the pathological hallmark of Parkinson's disease (PD) and dementia with Lewy bodies (DLB), but is also found in Alzheimer disease (AD). Therefore, there is a gaining interest in α-synuclein in cerebrospinal fluid (CSF) as potential biomarker for these neurodegenerative diseases. To broaden the available choices of α-synuclein measurement in CSF, we developed and validated a new assay for detecting total α-synuclein.

METHODS

This novel ELISA uses commercially available antibodies and is based on electrochemiluminescence technology. The assay protocol is straightforward, with short and simple incubation steps, and requires only small amounts of CSF. We validated this assay for precision, parallelism, dilution linearity, specificity, and spike recovery. We further compared it to the newly validated α-synuclein assay from BioLegend by analyzing a set of 50 CSF samples with both assays.

RESULTS

The new assay quantifies α-synuclein in CSF with a lower limit of detection of 36.3 pg/mL and shows no cross-reactivity with human β- and γ-synuclein. Results of dilution linearity, parallelism, spike recovery, and precision classify this assay as well suited for α-synuclein detection in human CSF samples.

CONCLUSIONS

We present a novel assay based on freely available components to quantify total α-synuclein in CSF as an additional method for α-synuclein as a biomarker in neurodegenerative diseases. The assay convinces with its simple and convenient protocol paired with high sensitivity.

摘要

背景

α-突触核蛋白异常聚集和沉积是帕金森病(PD)和路易体痴呆(DLB)的病理标志,但在阿尔茨海默病(AD)中也有发现。因此,脑脊液(CSF)中的α-突触核蛋白作为这些神经退行性疾病的潜在生物标志物越来越受到关注。为了拓宽脑脊液中α-突触核蛋白测量的可用选择,我们开发并验证了一种检测总α-突触核蛋白的新方法。

方法

这种新型酶联免疫吸附测定(ELISA)使用市售抗体,基于电化学发光技术。检测方案简单明了,孵育步骤简短,仅需少量脑脊液。我们对该方法的精密度、平行性、稀释线性、特异性和加标回收率进行了验证。我们通过用这两种方法分析一组50份脑脊液样本,进一步将其与BioLegend新验证的α-突触核蛋白检测方法进行比较。

结果

新方法对脑脊液中的α-突触核蛋白进行定量,检测下限为36.3 pg/mL,与人β-突触核蛋白和γ-突触核蛋白无交叉反应。稀释线性、平行性、加标回收率和精密度的结果表明,该方法非常适合检测人脑脊液样本中的α-突触核蛋白。

结论

我们提出了一种基于免费可用组件的新型检测方法,用于定量脑脊液中的总α-突触核蛋白,作为神经退行性疾病中α-突触核蛋白作为生物标志物的另一种方法。该检测方法以其简单方便的方案和高灵敏度令人信服。

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