采用质谱法对脑脊髓液淀粉样蛋白-β 进行定量测量。
Quantitative Measurement of Cerebrospinal Fluid Amyloid-β Species by Mass Spectrometry.
机构信息
Department of Neurology, Hirosaki National Hospital, Hirosaki, Aomori, Japan.
Department of Neurology, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
出版信息
J Alzheimers Dis. 2021;79(2):573-584. doi: 10.3233/JAD-200987.
BACKGROUND
High sensitivity liquid chromatography mass spectrometry (LC-MS/MS) was recently introduced to measure amyloid-β (Aβ) species, allowing for a simultaneous assay that is superior to ELISA, which requires more assay steps with multiple antibodies.
OBJECTIVE
We validated the Aβ1-38, Aβ1-40, Aβ1-42, and Aβ1-43 assay by LC-MS/MS and compared it with ELISA using cerebrospinal fluid (CSF) samples to investigate its feasibility for clinical application.
METHODS
CSF samples from 120 subjects [8 Alzheimer's disease (AD) with dementia (ADD), 2 mild cognitive dementia due to Alzheimer's disease (ADMCI), 14 cognitively unimpaired (CU), and 96 neurological disease subjects] were analyzed. Aβ species were separated using the Shimadzu Nexera X2 system and quantitated using a Qtrap 5500 LC-MS/MS system. Aβ1-40 and Aβ1-42 levels were validated using ELISA.
RESULTS
CSF levels in CU were 666±249 pmol/L in Aβ1-38, 2199±725 pmol/L in Aβ1-40, 153.7±79.7 pmol/L in Aβ1-42, and 9.78±4.58 pmol/L in Aβ1-43. The ratio of the amounts of Aβ1-38, Aβ1-40, Aβ1-42, and Aβ1-43 was approximately 68:225:16:1. Linear regression analyses showed correlations among the respective Aβ species. Both Aβ1-40 and Aβ1-42 values were strongly correlated with ELISA measurements. No significant differences were observed in Aβ1-38 or Aβ1-40 levels between AD and CU. Aβ1-42 and Aβ1-43 levels were significantly lower, whereas the Aβ1-38/1-42, Aβ1-38/1-43, and Aβ1-40/Aβ1-43 ratios were significantly higher in AD than in CU. The basic assay profiles of the respective Aβ species were adequate for clinical usage.
CONCLUSION
A quantitative LC-MS/MS assay of CSF Aβ species is as reliable as specific ELISA for clinical evaluation of CSF biomarkers for AD.
背景
液相色谱-质谱联用（LC-MS/MS）技术最近被用于检测淀粉样蛋白-β（Aβ）的各种亚型，该方法同时进行检测，优于需要使用多种抗体进行多个步骤检测的 ELISA 法。
目的
我们使用 LC-MS/MS 技术对 Aβ1-38、Aβ1-40、Aβ1-42 和 Aβ1-43 进行了验证，并与 ELISA 法检测脑脊液（CSF）样本进行了比较，以探讨其在临床应用中的可行性。
方法
分析了 120 例受试者（8 例痴呆型阿尔茨海默病（AD），2 例轻度认知障碍的 AD 患者（ADMCI），14 例认知正常的个体（CU）和 96 例神经科疾病患者）的 CSF 样本。采用岛津 Nexera X2 系统分离 Aβ 各亚型，使用 Qtrap 5500 LC-MS/MS 系统定量。使用 ELISA 法对 Aβ1-40 和 Aβ1-42 水平进行验证。
结果
CU 组 CSF 中 Aβ1-38 的水平为 666±249 pmol/L，Aβ1-40 为 2199±725 pmol/L，Aβ1-42 为 153.7±79.7 pmol/L，Aβ1-43 为 9.78±4.58 pmol/L。Aβ1-38、Aβ1-40、Aβ1-42 和 Aβ1-43 的比例约为 68:225:16:1。线性回归分析显示各 Aβ 亚型之间存在相关性。Aβ1-40 和 Aβ1-42 值与 ELISA 测量结果具有强相关性。AD 组和 CU 组之间 Aβ1-38 或 Aβ1-40 水平无显著差异。AD 组 Aβ1-42 和 Aβ1-43 水平显著降低，而 Aβ1-38/1-42、Aβ1-38/1-43 和 Aβ1-40/Aβ1-43 比值显著升高。各 Aβ 亚型的基本检测谱适用于临床应用。
结论
LC-MS/MS 定量检测 CSF 中 Aβ 各亚型与特异性 ELISA 检测 CSF 生物标志物用于 AD 的临床评估同样可靠。
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