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

描述(由申请人提供)：这项STTR提案的目标是开发一种B型利钠肽(BNP)的二维扩散点检测(POC)。这项拟议工作的主要目标是创建一种测试形式，使慢性心力衰竭患者能够 定量自我监测血清BNP水平。BNP基线水平的测定及其随时间的变化对表现为急性冠脉综合征和急性心力衰竭的患者具有预后意义，数据表明，随着时间的推移监测水平可能是通过慢性监测心力衰竭患者的容量状态并潜在地指导他们的治疗来进行传统临床评估的有用辅助工具。虽然BNP的车点检测在门诊环境中的应用是显而易见的，但定量设备仍然很繁琐，不太适合患者参与他们自己的监测和护理。在这项提案中，我们利用技术进步来简化实时测试和数据传输，以便将测试交到患者手中。这将通过使用2-D扩散分析平台来实现，该平台由一个纳米级的“无污染”聚甲基丙烯酸寡乙二醇酯刷涂玻璃芯片组成，其中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”微点。一根手指的血液被应用到芯片上，芯片中的蛋白质分析物 血液在无污垢的刷子上扩散，并与嵌入刷子内的捕获抗体的稳定斑点结合。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探测器进行的，该探测器连接到Android智能手机上，并使用手机的摄像头来记录荧光微点的图像。一个内置的手机应用程序可以将荧光点的强度转换为分析物浓度。图像被自动上传到安全的中央服务器，该服务器允许在自动更新患者的电子健康记录之前对图像进行第二次分析。该分析结合了一个内置的校准方案，能够最大限度地减少POC测试中固有的许多采样变量。此外，可以通过肉眼通过定制检测器查看微阵列，并在智能手机不可用时与校准斑点进行视觉比对。

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.