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的便携式分析仪和可行性 使用此平台技术执行多重免疫测定。多重能力成为可能 通过使用小的电子p-chip。最初的概念验证申请将是一个免疫测定法 血清或血浆样品中对登革热病毒（DENV）抗体的浓度。测定将是 旨在快速确定患者是否以前已被四个病毒之一感染 血清型。此信息至关重要，因为对DENV的先前免疫会在疾病上急剧恶化 重新暴露于异源病毒血清型后，进展并最终导致疾病结果。长期 目标是为多重免疫测定提供便携式测定平台，该平台分析了关键的生物标志物 多种疾病。该系统的优点是快速评估速度（从图 患者的血液样本以报告结果），两者的样本处理最少和低成本 分析仪和消耗品。 电子芯片是小型的激光激活微转换器（“ p-Chips”），很小（600 µm x 600 µm x 100 µm），化学惰性和高度稳定。 p-chip，每个芯片都包含一个唯一的序列号，是 与特定蛋白质生物标志物的抗体抗体有关，然后与A孵育 样品并用荧光试剂检测到结合。 P-Chips的关键特征是它们是被动的 只有直接用脉冲光照明的设备才能变得活跃，从而使它们能够成为 在闭合的旋转管中旋转时，会单独读取，暴露于集中的激光束。这 读数依赖于荧光强度和芯片序列号的简单确定 （ID），标识结合探针。多重测定是通过在 在同一试管中的P-CHIP以一种容易读取数十个数百的试管的方式 P-CHIP和报告会导致很短的时间。该项目的主要目标是（1）建造一个 原型读取器和开发将控制设备并分析数据的软件； （2） 演示并表征用于检测登革热病毒的单管一步无洗的免疫测定法。 提议的方法的结果将是一个新的系统，用于测试患者的暴露 最近或过去，到登革热病毒。该系统将由便携式分析仪（“ Cyclone”）组成 以及在P-CHIP上配置的测定，量化了血清中抗登记ABS的四个血清型。系统 可以轻松扩展以包括其他感兴趣的生物标志物。