POC 2-D Diffusion Assay for self-monitoring of BNP

描述（由申请方提供）：本STTR提案的目的是开发B型利钠肽（BNP）的二维扩散床旁检测（POC）。拟议工作的主要目标是创建一种测试格式，使慢性心力衰竭患者能够 定量自我监测BNP的血清水平。BNP基线水平的测定和该水平随时间的变化对急性冠状动脉综合征和急性心力衰竭患者具有预后效用，数据表明，通过长期监测心力衰竭患者的容量状态，监测随时间的水平可能是传统临床评价的有用辅助手段，并可能指导其治疗。虽然BNP的车点测试在门诊诊所环境中的应用是显而易见的，但定量设备仍然很麻烦，并且不太适合患者参与自己的监测和护理。在这项提案中，我们利用技术进步，简化了实时测试和数据传输，以便将测试放在患者手中。这将通过使用2-D扩散测定平台来实现，该平台由纳米级“无污染”聚（低聚乙二醇甲基丙烯酸酯）刷涂玻璃芯片组成，该芯片包含两种类型的印刷微点：捕获抗体的“稳定”微点和检测试剂的“可溶性”微点。将手指针刺的血液施加到芯片上，并且在芯片中的蛋白质分析物被检测。 血液扩散穿过无污垢刷并结合到嵌入刷内的捕获抗体的稳定点。同时，血液溶解荧光标记的检测抗体的可溶性斑点，使得检测抗体可以扩散并标记由捕获抗体的稳定斑点结合的任何分析物。检测是通过一个定制的荧光检测器，连接到Android智能手机，并使用手机的相机记录荧光微点的图像。通过内置的手机应用程序将荧光点强度转换为分析物浓度。图像自动上传到安全的中央服务器，在自动更新患者的电子健康记录之前，允许对图像进行第二次分析。该检测试剂盒包含一个内置的校准方案，能够最大限度地减少POC检测中固有的许多采样变量。此外，微阵列可以通过定制的检测器用肉眼观察，并且当智能手机不可用时，可以与校准点并排进行视觉比较。

DESCRIPTION (provided by applicant): The objective of this STTR proposal is to develop a 2-D diffusion point-of-care assay (POC) for B-type natriuretic peptide (BNP). The primary goal of the proposed work is to create a test format that will allow patients with chronic heart failure to quantitatively self-monitor serum levels of BNP. Determination of baseline level of BNP and changes in this level over time have prognostic utility in patients presenting with acute coronary syndromes and acute heart failure, and data suggest that monitoring levels over time may be a useful adjunct to traditional clinical evaluation by chronic monitoring of volume status in heart failure patients and potentially in guiding their treatment. While the applications of point-of-car testing of BNP in the outpatient clinic setting are obvious, quantitative devices remain cumbersome and are not well suited to patient engagement in their own monitoring and care. In this proposal, we exploit technological advances that simplify real-time testing and data transmission in order to place testing in the hands of patients. This will be accomplished through the use of the 2-D diffusion assay platform, which consists of a nanoscale "nonfouling" poly(oligoethylene glycol methacrylate) brush coated glass chip that contains two types of printed microspots: "stable" microspots of capture antibodies and "soluble" microspots of detection reagents. A finger stick of blood is applied to the chip, and the protein analytes in the blood diffuse across the non-fouling brush and bind to stable spots of capture antibodies embedded within the brush. Simultaneously, the blood dissolves the soluble spots of fluorescently labeled detection antibodies, so that the detection antibodies can diffuse and label any analyte bound by the stable spots of capture antibodies. Detection is via a custom fluorescence detector that attaches to an Android smart phone and uses the phone's camera to record an image of the fluorescent microspots. Fluorescent spot intensities are converted to analyte concentration by a built in phone app. Images are automatically uploaded to a secure central server that allows a second analysis of the image to take place before automatically updating the patient's electronic health record. The assay incorporates a built in calibration scheme capable of minimizing many of the sampling variables inherent in POC testing. In addition, the microarray can be viewed through the custom detector by the naked eye and visually compared side-by-side with calibration spots when a smartphone is not available.