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）：阿尔茨海默病（AD）是一种难以诊断和治疗的复杂疾病，是痴呆症最常见的形式。目前，没有现成的生物标志物测试小组来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还没有得到充分的证明。该项目的目标是为阿尔茨海默病患者的血液样本开发一种多重生物标志物测试。在该资助的第一阶段，我们将展示一种新的快速多重检测阿尔茨海默病血清/血浆生物标志物的可行性。我们开发了一种专有的方法，使用实时聚合酶链反应（PCR）技术以多路复用的方式测量蛋白质浓度，具有非常高的灵敏度和重复性。该方法，多重实时免疫pcr (MRI-PCR)，包括一个三明治免疫测定，其中检测抗体被标记为不同的DNA序列。实时聚合酶链式反应（Real-time PCR）用于同时定量所有DNA标记，从而揭示生物样品中同源蛋白的数量。初步数据表明，与标准的ELISA检测相比，采用MRI-PCR检测的多重细胞因子检测灵敏度提高了约2个数量级，动态范围增加了约2个数量级，并且具有非常低的标准偏差（CV <10%）和高特异性。在I期研究中，将准备一组试剂来检测15种AD的人类生物标志物，并在MRI-PCR分析中进行评估。然后将目标蛋白加入到正常的人血清中，以在生理样品中验证该分析。需要评估的参数包括灵敏度、动态范围、准确度、精密度、剂量-响应线性和特异性。将进行一项小型试点研究，以评估该测试在AD患者血清样本以及年龄匹配的正常对照（n=10-20个样本）中的性能。在II期，该检测将在大规模临床研究中进一步验证，以证明该技术在阿尔茨海默病的诊断、预后和治疗监测方面的价值。这些研究的结果将导致一种比脑脊液分析侵入性更小的AD新测试，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的检测方法，以协助阿尔茨海默病的诊断、预后和治疗监测。