Veitinger Michael, Oehler Rudolf, Umlauf Ellen, Baumgartner Roland, Schmidt Georg, Gerner Christopher, Babeluk Rita, Attems Johannes, Mitulovic Goran, Rappold Eduard, Lamont John, Zellner Maria
Center of Physiology and Pharmacology, Institute of Physiology, Medical University of Vienna, Schwarzspanierstrasse 17, 1090, Vienna, Austria.
Acta Neuropathol. 2014 Nov;128(5):665-77. doi: 10.1007/s00401-014-1341-8. Epub 2014 Sep 24.
Alzheimer's disease (AD), a multifactorial neurodegenerative condition caused by genetic and environmental factors, is diagnosed using neuropsychological tests and brain imaging; molecular diagnostics are not routinely applied. Studies have identified AD-specific cerebrospinal fluid (CSF) biomarkers but sample collection requires invasive lumbar puncture. To identify AD-modulated proteins in easily accessible blood platelets, which share biochemical signatures with neurons, we compared platelet lysates from 62 AD, 24 amnestic mild cognitive impairment (aMCI), 13 vascular dementia (VaD), and 12 Parkinson's disease (PD) patients with those of 112 matched controls by fluorescence two-dimensional differential gel electrophoresis in independent discovery and verification sets. The optimal sum score of four mass spectrometry (MS)-identified proteins yielded a sensitivity of 94 % and a specificity of 89 % (AUC = 0.969, 95 % CI = 0.944-0.994) to differentiate AD patients from healthy controls. To bridge the gap between bench and bedside, we developed a high-throughput multiplex protein biochip with great potential for routine AD screening. For convenience and speed of application, this array combines loading control-assisted protein quantification of monoamine oxidase B and tropomyosin 1 with protein-based genotyping for single nucleotide polymorphisms (SNPs) in the apolipoprotein E and glutathione S-transferase omega 1 genes. Based on minimally invasive blood drawing, this innovative protein biochip enables identification of AD patients with an accuracy of 92 % in a single analytical step in less than 4 h.
阿尔茨海默病（AD）是一种由遗传和环境因素引起的多因素神经退行性疾病，通过神经心理学测试和脑成像进行诊断；分子诊断方法并未常规应用。研究已鉴定出AD特异性脑脊液（CSF）生物标志物，但样本采集需要进行侵入性腰椎穿刺。为了在易于获取的血小板中鉴定受AD调节的蛋白质，血小板与神经元具有共同的生化特征，我们通过荧光二维差异凝胶电泳，在独立的发现和验证组中，将62例AD患者、24例遗忘型轻度认知障碍（aMCI）患者、13例血管性痴呆（VaD）患者和12例帕金森病（PD）患者的血小板裂解物与112例匹配对照的血小板裂解物进行了比较。四种经质谱（MS）鉴定的蛋白质的最佳总分在区分AD患者与健康对照时，灵敏度为94%，特异性为89%（曲线下面积[AUC]=0.969，95%置信区间[CI]=0.944 - 0.994）。为了弥合实验室研究与临床应用之间的差距，我们开发了一种具有很大潜力用于AD常规筛查的高通量多重蛋白质生物芯片。为方便应用并提高速度，该阵列将单胺氧化酶B和原肌球蛋白1的加载对照辅助蛋白质定量与载脂蛋白E和谷胱甘肽S-转移酶ω1基因单核苷酸多态性（SNP）的基于蛋白质的基因分型相结合。基于微创采血，这种创新的蛋白质生物芯片能够在不到4小时的单个分析步骤中，以92%的准确率识别AD患者。
