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

总结 本课题的目的是论证一种新型的便携式分析仪的可行性， 利用该平台技术进行多重免疫测定。使多路复用能力成为可能 通过使用小的电子p芯片最初的概念验证应用将是一种免疫测定， 血清或血浆样本中登革热病毒（DENV）抗体的浓度。对试剂盒进行 旨在快速确定患者是否曾感染过四种病毒之一， 血清型这一信息是至关重要的，因为预先存在的对DENV的免疫力会使疾病急剧恶化。 疾病进展和最终疾病结果。长期 本发明的目的是提供一种用于多重免疫测定的便携式测定平台， 各种疾病。该系统的优点是快速的测定速度（从绘制一个样品开始的周转时间）。 病人的血液样本的结果报告），最小的样品处理和低成本的两个 分析仪和耗材。 电子芯片是激光激活的微型应答器（“p-Chips”），其尺寸较小（600 µm x 600 µm x 100 µm）， 100 µm），化学惰性且高度稳定。p芯片，每个芯片都包含一个唯一的序列号， 用探针衍生化，在这种情况下是特异性蛋白质生物标志物的抗体，然后用 样品和结合用荧光试剂检测。p-Chips的一个关键特征是它们是被动的 只有当直接用脉冲光照射时才能激活的器件，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。的 读出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。通过将不同的探针放置在 p-芯片在同一个试管中，以一种容易读取阅读数十到数百个的方式， p-Chips和报告结果在很短的时间内。该项目的主要目标是（1）建立一个 原型阅读器和开发软件，既控制设备，又分析数据;和（2） 展示和表征用于检测登革病毒的一管一步免洗涤免疫测定。 所提出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，登革热病毒。该系统将由便携式分析仪（“Cyclone”） 以及配置在p-Chips上的测定法，其定量血清中抗登革热Ab的四种血清型。系统 可以容易地扩展到包括感兴趣的其他生物标志物。

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.