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通过减少磁珠数量提高单分子阵列免疫检测技术（Simoa）的灵敏度，有助于对干血斑中磷酸化tau蛋白181位点（pTau-181）进行定量分析。

Increasing the sensitivity of Simoa via bead count reduction facilitates the quantification of pTau-181 in dried plasma spots.

作者信息

Mohaupt Pablo, Vialaret Jérôme, Hirtz Christophe, Lehmann Sylvain

机构信息

LBPC-PPC Université de Montpellier IRMB CHU de Montpellier INM INSERM Montpellier France.

出版信息

Alzheimers Dement (N Y). 2024 Mar 11;10(1):e12456. doi: 10.1002/trc2.12456. eCollection 2024 Jan-Mar.

DOI:10.1002/trc2.12456
PMID:38476928
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10927909/
Abstract

INTRODUCTION

The exclusion of affected populations from Alzheimer's disease (AD) clinical research limits our understanding of disease heterogeneity and its impact on clinical care. While micro sampling with dried plasma spots (DPS) can promote inclusivity by enabling sample collection in remote areas, current techniques lack the sensitivity required for the quantification of phosphorylated tau at Thr181 (pTau-181) in DPS extracts.

METHODS

We developed an assay for pTau-181 with reduced bead count and improved bead read efficiency (BRE) using a prototype Simoa instrument. This novel assay's performance was evaluated against standard pTau-181 assays on two Simoa platforms, and DPS extracts were tested for pTau-181 quantification feasibility.

RESULTS

The novel assay quantifies pTau-181 at concentrations up to 16x lower than traditional pTau-181 assays on HD-X and SR-X platforms. DPS extracts tested with our low-bead assay were quantified considerably above the lower limit of quantification (LLOQ), indicating the suitability of this assay for future DPS extract measurements.

DISCUSSION

Implementing DPS sampling and pTau-181 quantification could increase participation from underrepresented groups in AD research. However, additional assay optimization and an in-depth study of preanalytical sample stability are essential for the transition to clinical applicability.

摘要

引言

将患病人群排除在阿尔茨海默病（AD）临床研究之外，限制了我们对疾病异质性及其对临床护理影响的理解。虽然使用干血斑（DPS）进行微量采样能够通过在偏远地区采集样本促进包容性，但目前的技术缺乏对DPS提取物中苏氨酸181位点磷酸化tau蛋白（pTau-181）进行定量所需的灵敏度。

方法

我们使用一台原型Simoa仪器开发了一种用于pTau-181的检测方法，该方法减少了磁珠数量并提高了磁珠读取效率（BRE）。在两个Simoa平台上，将这种新检测方法的性能与标准pTau-181检测方法进行了比较，并测试了DPS提取物进行pTau-181定量的可行性。

结果

与HD-X和SR-X平台上的传统pTau-181检测方法相比，这种新检测方法能够在低至16倍的浓度下对pTau-181进行定量。用我们的低磁珠检测方法测试的DPS提取物的定量结果大大高于定量下限（LLOQ），表明该检测方法适用于未来的DPS提取物测量。

讨论

实施DPS采样和pTau-181定量可以增加代表性不足群体参与AD研究的程度。然而，要向临床应用过渡，还需要进一步优化检测方法，并深入研究分析前样本稳定性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/ce1d12e56a6a/TRC2-10-e12456-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/5b5294017409/TRC2-10-e12456-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/1b5ee68fc696/TRC2-10-e12456-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/38038941b231/TRC2-10-e12456-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/66e3362e4a9b/TRC2-10-e12456-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/ce1d12e56a6a/TRC2-10-e12456-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/5b5294017409/TRC2-10-e12456-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/1b5ee68fc696/TRC2-10-e12456-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/38038941b231/TRC2-10-e12456-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/66e3362e4a9b/TRC2-10-e12456-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e8d/10927909/ce1d12e56a6a/TRC2-10-e12456-g003.jpg

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