Development and analytical validation of AB40, AB42, A-Syn, CD33, P-Tau 181, Tau, and TREM2 in human serum, plasma, or CSF

IF 13 1区医学 Q1 CLINICAL NEUROLOGY Alzheimer's & Dementia Pub Date : 2025-01-09 DOI:10.1002/alz.089257

Jacqueline Surls, Robyn Vega Ibanez, Lindsey Brown, Josh Kemp

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Abstract

Background

Measurement of blood-based biomarkers for Alzheimer disease is simpler and more accessible when compared to CSF. We report the development and validation of seven Simoa® immunoassays for the detection of AB40, AB42, A-Syn, CD33, P-Tau 181, Tau, and TREM2 in human serum, plasma, or CSF.

Method

Using the Quanterix® Simoa® technology (SR-X) platform, AB40, AB42, A-Syn, CD33, P-Tau 181, Tau, and TREM2 were developed and analytically validated in human serum, plasma, or CSF per CLSI standards.

Result

We have developed and analytically validated Simoa® immunoassays to detect AB40, AB42, A-Syn, CD33, P-Tau 181, Tau, and TREM2 in human serum, plasma, or CSF. The assays met acceptance criteria for sensitivity, precision, stability, interference, and parallelism. The calibration curves in serum/plasma were linear from 16-2000 pg/mL for AB40, 18-40,000 pg/mL for AB42, 0.13-80 ng/mL for A-Syn, 0.012-60 ng/mL for CD33, 0.60-3000 pg/mL for P-Tau 181, 0.12-600 pg/mL for Tau, and 0.30-38 ng/mL for TREM2. For CSF, the linear ranges were from 626-80,000 pg/mL for AB40, 72-160,000 pg/mL for AB42, 0.013-8.0 ng/mL for A-Syn, 0.012-60 ng/mL for CD33, 30-150,000 pg/mL for P-Tau 181, 0.30-1500 pg/mL for Tau, and 0.60-76 ng/mL for TREM2. The three quality controls met inter assay precision with %CV ≤16% for levels 1 and 2, and ≤20% for level 3 across all analytes. No interference was observed with bilirubin, hemoglobin, or triglyceride. The sensitivity of the assays in human serum/plasma was demonstrated by the LLOQ values of 28 pg/mL, 47 pg/mL, 0.20 ng/mL, 0.11 ng/mL, 2.8 pg/mL, 0.22 pg/mL, and 0.59 ng/mL for AB40, AB42, A-Syn, CD33, P-Tau 181, Tau, and TREM2, respectively. For human CSF, the LLOQ values of 1139 pg/mL, 187 pg/mL, 0.020 ng/mL, 0.11 ng/mL, 142 pg/mL, 0.56 pg/mL, and 1.2 ng/mL for AB40, AB42, A-Syn, CD33, P-Tau 181, Tau, and TREM2 in human CSF, respectively. The stability of the analytes was established under various conditions, such as freeze-thaw cycles, short-term and long-term storage.

Conclusion

These assays demonstrated acceptable performance for clinical implementation as research use only and may be useful for blood-based biomarker studies in clinical trials.

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AB40、AB42、A‐Syn、CD33、P‐Tau 181、Tau和TREM2在人血清、血浆或CSF中的开发和分析验证

与脑脊液相比，测量阿尔茨海默病的血液生物标志物更简单，更容易获得。我们报告了七种Simoa®免疫测定方法的开发和验证，用于检测人血清、血浆或CSF中的AB40、AB42、A‐Syn、CD33、P‐Tau 181、Tau和TREM2。方法使用Quanterix®Simoa®技术（SR - X）平台，开发AB40、AB42、A‐Syn、CD33、P‐Tau 181、Tau和TREM2，并根据CLSI标准在人血清、血浆或CSF中进行分析验证。结果我们开发并分析验证了Simoa®免疫测定法，用于检测人血清、血浆或CSF中的AB40、AB42、A‐Syn、CD33、P‐Tau 181、Tau和TREM2。该方法在灵敏度、精密度、稳定性、干扰性和平行性方面均符合验收标准。血清/血浆的校准曲线为线性，AB40为16‐2000 pg/mL， AB42为18‐40000 pg/mL， A‐Syn为0.13‐80 ng/mL， CD33为0.012‐60 ng/mL， P‐Tau 181为0.60‐3000 pg/mL， Tau为0.12‐600 pg/mL， TREM2为0.30‐38 ng/mL。对于脑脊液，AB40的线性范围为626 ~ 80000 pg/mL， AB42的线性范围为72 ~ 160,000 pg/mL， A‐Syn的线性范围为0.013 ~ 8.0 ng/mL， CD33的线性范围为0.012 ~ 60 ng/mL， P‐Tau 181的线性范围为30 ~ 150000 pg/mL， Tau的线性范围为0.30 ~ 1500 pg/mL， TREM2的线性范围为0.60 ~ 76 ng/mL。在所有分析物中，3种质量控制均满足测定间精度，水平1和水平2的%CV≤16%，水平3的%CV≤20%。未观察到胆红素、血红蛋白或甘油三酯的干扰。AB40、AB42、A‐Syn、CD33、P‐Tau 181、Tau和TREM2的LLOQ值分别为28 pg/mL、47 pg/mL、0.20 ng/mL、0.11 ng/mL、2.8 pg/mL、0.22 pg/mL和0.59 ng/mL，证明了该检测方法在人血清/血浆中的敏感性。人脑脊液中AB40、AB42、A‐Syn、CD33、P‐Tau 181、Tau和TREM2的LLOQ值分别为1139 pg/mL、187 pg/mL、0.020 ng/mL、0.11 ng/mL、142 pg/mL、0.56 pg/mL和1.2 ng/mL。分析物在冻融循环、短期和长期储存等条件下的稳定性。结论：这些检测方法在临床应用中表现出可接受的性能，仅作为研究用途，可能对临床试验中基于血液的生物标志物研究有用。

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来源期刊

Alzheimer's & Dementia 医学-临床神经学

CiteScore

14.50

自引率

5.00%

发文量

299

审稿时长

3 months

期刊介绍： Alzheimer's & Dementia is a peer-reviewed journal that aims to bridge knowledge gaps in dementia research by covering the entire spectrum, from basic science to clinical trials to social and behavioral investigations. It provides a platform for rapid communication of new findings and ideas, optimal translation of research into practical applications, increasing knowledge across diverse disciplines for early detection, diagnosis, and intervention, and identifying promising new research directions. In July 2008, Alzheimer's & Dementia was accepted for indexing by MEDLINE, recognizing its scientific merit and contribution to Alzheimer's research.