Novel Multiplex Assay for Biomarkers of Alzheimer's Disease

阿尔茨海默病生物标志物的新型多重检测

• 批准号: 7745372
• 负责人: SUSAN C WRIGHT
• 金额: $ 16.17万
• 依托单位: PANORAMA RESEARCH, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7745372
• 关键词: Age; Alzheimer' s Disease; Antibodies; Biological; Biological Assay; Biological Markers; Blood; Blood specimen; Cerebrospinal Fluid; Clinical Research; Complex; DNA; DNA Sequence; Data; Dementia; Detection; Diagnosis; Diagnostic; Disease; Disease Progression; Dose; Enzyme-Linked Immunosorbent Assay; Evaluation; Goals; Grant; Human; Label; Lead; Measurement; Measures; Methods; Monitor; Parkinson Disease; Patients; Phase; Phase I Clinical Trials; Physiological; Pilot Projects; Plasma; Polymerase Chain Reaction; Proteins; Reagent; Reproducibility; Research Personnel; Sampling; Sensitivity and Specificity; Serum; Specificity; Technology; Testing; Time; Validation; cytokine; minimally invasive; novel; outcome forecast; performance tests; public health relevance; response

DESCRIPTION (provided by applicant): Alzheimer's disease (AD), the most common form of dementia, is a complex disorder that is difficult to diagnose and treat. Currently, there is no readily available biomarker test panel to aid diagnosis or monitor disease progression. Although analysis of biomarkers in cerebrospinal fluid (CSF) has provided useful information, the validation of biomarkers in blood has not been as well-documented. The goal of this project is to develop a multiplex biomarker test for blood samples from Alzheimer's patients. In Phase I of this grant, we will demonstrate the feasibility of a novel rapid multiplex assay for serum/plasma biomarkers of AD. We have developed a proprietary method for using the real-time-polymerase chain reaction (PCR) technology to measure protein concentrations in a multiplexed fashion with very high sensitivity and reproducibility. The method, Multiplexed Real-Time-Immuno-PCR (MRI-PCR), consists of a sandwich immuno-assay in which the detection antibodies are labeled with different DNA sequences. Real-time PCR is then used to quantify all of the DNA labels in parallel, thus revealing the quantities of the cognate proteins in the biological samples. Preliminary data have demonstrated that multiplexed cytokine measurements employing MRI-PCR assays provides ~2 orders of magnitude more sensitive detection and >2 orders of magnitude increased dynamic range in comparison to standard ELISA assays, along with very low standard deviations (CV <10%) and high specificity. In Phase I studies, a panel of reagents to detect 15 human biomarkers of AD will be prepared and evaluated in the MRI-PCR assay. The target proteins will then be spiked into normal human serum to validate the assay in a physiological sample. Parameters to be evaluated include sensitivity, dynamic range, accuracy, precision, dose-response linearity, and specificity. A small pilot study will be conducted to evaluate the performance of the test on serum samples from AD patients as well as age-matched normal controls (n=10-20 samples). In Phase II, the assay will be further validated in a large scale clinical study to demonstrate the value of this technology in the diagnosis, prognosis and monitoring of therapy for AD. The results of these studies will lead to a new test for AD that is less invasive than CSF analysis and will provide valuable information to both clinicians as well as researchers. PUBLIC HEALTH RELEVANCE: This project will develop a novel assay to assist in the diagnosis, prognosis, and monitoring of therapy for Alzheimer's disease.

描述（由申请人提供）：阿尔茨海默氏病（AD）是痴呆症的最常见形式，是一种很难诊断和治疗的复杂疾病。目前，没有容易获得的生物标志物测试面板可以帮助诊断或监测疾病进展。尽管对脑脊液（CSF）中生物标志物的分析提供了有用的信息，但血液中生物标志物的验证尚未得到充分证明。该项目的目的是为阿尔茨海默氏症患者的血液样本开发多重生物标志物测试。在该赠款的第一阶段，我们将证明一种新型的快速多重分析对AD的血清/等离子体生物标志物的可行性。我们开发了一种专有方法，用于使用实时聚合酶链反应（PCR）技术以非常高灵敏度和可重复性的多重方式测量蛋白质浓度。该方法是多重的实时免疫PCR（MRI-PCR），由三明治免疫测定组成，其中检测抗体用不同的DNA序列标记。然后使用实时PCR并行量化所有DNA标记，从而揭示了生物样品中同源蛋白的量。初步数据表明，使用MRI-PCR分析的多路复用细胞因子测量值可提供约2个数量级的更敏感的检测顺序，与标准ELISA分析相比，动态范围更高，> 2个数量级，以及非常低的标准偏差（CV <10％）和高特异性。在第一阶段的研究中，将在MRI-PCR分析中制备和评估一组试剂，以检测15种AD的人类生物标志物。然后，靶蛋白将被刺激到正常的人血清中，以验证生理样品中的测定。要评估的参数包括灵敏度，动态范围，准确性，精度，剂量反应线性和特异性。将进行一项小型试点研究，以评估AD患者的血清样品以及年龄匹配的正常对照（n = 10-20个样品）的测试性能。在第二阶段，该测定法将在大规模的临床研究中得到进一步验证，以证明该技术在诊断，预后和监测AD治疗中的价值。这些研究的结果将导致对AD的新测试，该测试比CSF分析不那么侵入性，并将为临床医生和研究人员提供有价值的信息。公共卫生相关性：该项目将开发一种新颖的测定法，以帮助诊断，预后和监测阿尔茨海默氏病的治疗。