Portable Bioassay System Based on Electronic p-Chips: Application to Detect Dengue Antibodies

基于电子芯片的便携式生物测定系统：用于检测登革热抗体

• 批准号: 10080000
• 负责人: WLODEK MANDECKI
• 金额: $ 14.99万
• 依托单位: PHARMASEQ, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10080000
• 关键词: Portable; Bioassay; System; Based; Electronic

Summary The purpose of the present project is to demonstrate a new type of portable analyzer and the feasibility of performing multiplex immunoassays utilizing this platform technology. The multiplex capability is made possible by using small, electronic p-Chips. The initial proof-of-concept application will be an immunoassay determining the concentration of antibodies to dengue virus (DENV) in serum or plasma samples. The assay will be designed to rapidly determine whether a patient has been previously infected with one of the four viral serotypes. This information is crucial as pre-existing immunity to DENV can drastically worsen disease progression and ultimately disease outcome upon re-exposure to heterologous viral serotypes. The long-term goal is to provide a portable assay platform for multiplex immunoassays that analyze key biomarkers for a variety of diseases. The advantages of this system are rapid assay speed (turnaround time from drawing a patient's blood sample to the reporting of the results), minimal sample handling and low cost for both the analyzer and consumables. The electronic chips are laser light-activated microtransponders ("p-Chips") that are small (600 µm x 600 µm x 100 µm), chemically inert and highly stable. p-Chips, each of which contains a unique serial number, are derivatized with probes, in this case antibodies to specific protein biomarkers, and then incubated with a sample and binding detected with fluorescent reagent(s). A key feature of p-Chips is that they are passive devices that become active only when directly illuminated with pulsed light, thus enabling them to be individually read while spinning in a closed, rotating tube exposed to a focused excitation laser beam. The readout relies on the simultaneous determination of the intensity of fluorescence and the chip serial number (ID), which identifies the binding probe. Multiplex assays are constructed by placing different probes on p-Chips within the same test tube in a manner that lends itself readily to reading dozens to hundreds of p-Chips and reporting results in a very short period of time. The main goals of this project are to (1) build a prototype reader and develop software that will both control the device and analyze the data; and (2) demonstrate and characterize a one-tube-one-step-no-wash immunoassay for detecting dengue virus. The results for the proposed approach will be a novel system for testing the exposure of a patient, either recently or in the past, to the dengue virus. The system will be composed of the portable analyzer ("Cyclone") and the assay configured on p-Chips quantifying four serotypes of the anti-dengue Abs in serum. The system can be expanded easily to include additional biomarkers of interest.

概括 本项目的目的是展示一种新型便携式分析仪及其可行性 利用该平台技术进行多重免疫测定使多重功能成为可能。 通过使用小型电子 p-芯片 最初的概念验证应用将是免疫分析测定。 血清或血浆样本中的登革热病毒 (DENV) 抗体的测定将是浓度。 旨在快速确定患者之前是否感染过四种病毒中的一种 该信息至关重要，因为预先存在的 DENV 免疫力可能会加剧疾病恶化。 重新暴露于异源病毒血清型时的进展和最终疾病结果。 目标是为多重免疫分析提供便携式分析平台，分析关键生物标志物 该系统的优点是检测速度快（从绘制结果开始的周转时间）。 患者的血液样本到报告结果），样本处理最少，成本低 分析仪和消耗品。 电子芯片是小型激光激活微转发器（“p-芯片”）（600 µm x 600 µm x 100 µm），化学惰性且高度稳定，每个 p-Chips 都包含唯一的序列号。 用探针衍生化，在这种情况下是针对特定蛋白质生物标志物的抗体，然后与 p-Chips 的一个关键特征是它们是被动的。 仅当直接用脉冲光照射时才会激活的设备，从而使它们能够 在暴露于聚焦激发激光束的封闭旋转管中旋转时单独读取。 读数依赖于荧光强度和芯片序列号的同时测定 (ID)，其识别结合探针是通过将不同的探针放置在其上构建的。 p-芯片位于同一试管内，可以轻松读取数十到数百个数据 p-Chips 并在很短的时间内报告结果 该项目的主要目标是 (1) 构建一个。 原型阅读器并开发控制设备和分析数据的软件；以及 (2) 展示并表征用于检测登革热病毒的一管一步免洗免疫测定法。 所提出方法的结果将是一个用于测试患者暴露情况的新颖系统， 最近或过去，针对登革热病毒的系统将由便携式分析仪（“旋风”）组成。 以及在 p-Chips 上配置的测定法，用于量化血清中抗登革热抗体的四种血清型。 可以轻松扩展以包含其他感兴趣的生物标志物。