PFI-TT: Clip-On Smartphone Biosensor for Mobile Health Diagnostics

创新伙伴关系-技术转化（PFI-TT）项目的更广泛影响/商业潜力是开发一种单步、30分钟的艾滋病毒载量检测方法，具有数字分辨率和测量少量病毒载量的能力。该系统是为没有实验室专业知识的艾滋病患者设计的，这在目前是不可能的。该测试将通过一种廉价的夹式仪器来测试血液样本；它使用任何现代智能手机的后置摄像头来记录化学反应的视频，并测量特殊的光信号。该测试将测量危险病原体，包括纠正假阳性和假阴性的方法；它还提供结果的即时显示，并与基于云的数据库共享数据。由于血液中的HIV传感是具有巨大商业市场的具有挑战性的应用，成功的演示将为许多潜在感染的移动传感的广泛产品和服务铺平道路。提出的项目是设计，构建并严格验证一个自我测试平台，用于在血清滴中进行艾滋病毒的数字分辨率定量。目前，没有一种自我检测方法能够在病毒血症高峰和急性感染的传播性期间检测出艾滋病毒。所提出的方法利用一种廉价的（10美元）夹式仪器，使任何现代智能手机的后置摄像头都能作为荧光显微镜，照亮并等温加热预加载有用于hiv特异性核酸序列环介导等温扩增（LAMP）引物的硅基测试盒。提出的技术将整合几个创新元素，实现30分钟内从样本到答案的测试目标，同时通过执行以下操作实现1000拷贝/ml的检测限：LAMP反应在低自荧光硅基微流控盒内预装干引物，并包装为无冷藏储存。2. 空间LAMP （S-LAMP）分析利用视频成像的过程和自动图像分析来识别/计数荧光发光的LAMP起始点。3. 整合阳性和阴性实验控制区，确保分析的有效性。4. 在将LAMP缓冲液和测试样品放入纸质测试介质之前，对样品进行化学裂解。5. 一种夹式仪器，与任何智能手机后置摄像头都有光学接口。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of the Partnership for Innovation - Technology Translation (PFI-TT) project is to develop a single-step, 30 minute test for HIV viral load measurement with digital resolution and the capability to measure small amounts. This system is designed to be used by HIV patients with no laboratory expertise, which is currently not possible. The test will be conducted by an inexpensive clip-on instrument to test a blood sample; it uses the rear-facing camera of any modern smartphone to record video of the chemical reaction and measure special light signals. The test will measure the dangerous pathogens, including a way to correct for false positives and negatives; it also provides immediate display of results and shares the data with a cloud-based database. As HIV sensing in blood represents a challenging application with a large commercial market, a successful demonstration will pave the way toward a host of a wide array products and services for mobile sensing of many potential infections. The proposed project is to design, build, and rigorously validate a self-testing platform for digital-resolution quantitation of HIV within a droplet of serum. Currently, no self-testing approaches are currently able to detect HIV during peak viremia and transmissibility in acute infection. The proposed approach utilizes an inexpensive ($10) clip-on instrument that will enable the back-facing camera of any modern smartphone to function as a fluorescence microscope that illuminates and isothermally heats a silicon-based test cartridge that is pre-loaded with primers for Loop-Mediated Isothermal Amplification (LAMP) of HIV-specific nucleic acid sequences. The proposed technology will integrate several innovative elements to achieve the goal of a sample-to-answer test within 30 minutes, while achieving 1000 copies/ml detection limit by performing: 1. LAMP reactions within a low-autofluorescence silicon-based microfluidic cartridge pre-loaded with dried primers, and packaged for refrigeration-free storage. 2. Spatial LAMP (S-LAMP) analysis of the amplification reaction using video imaging of the process and automated image analysis to identify/count fluorescent-emitting LAMP initiation points. 3. Integration of positive and negative experimental control regions to ensure assay validity. 4. Chemical lysis of the sample prior to dispensing of LAMP buffer and the test sample into the paper test medium. 5. A clip-on instrument that makes an optical interface with any smartphone rear-facing camera.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.