Smartphone-based diagnostic for HIV self-testing

摘要 联合国艾滋病规划署认可的90-90-90目标建议诊断全球90%的艾滋病毒感染者， 让90%的人接受有效的抗逆转录病毒治疗，并确保90%的治疗者实现 到2020年持续抑制病毒载量。不幸的是，美国没有准备好实现这一目标;只有 在美国，30%的HIV感染者目前实现了病毒载量抑制。显然，一个范式的转变， 要达到这个目标，必须进行病毒载量测试。该提案旨在创建一个定量的，手持的， 病毒载量自我测试，可由患者自己使用，以支持他们的管理这种慢性 该系统能够为远程医疗保健提供者提供相关的临床见解。 在我们前期工作的基础上，R61阶段的目标是将联合收割机等温环- 介导的核酸扩增（LAMP），具有高灵敏度和无标记的读出方法，称为 在手持式平台上以最少的工艺步骤定量HIV病毒载量的粒子扩散法。 具体的里程碑包括：1）优化LAMP检测，以确保最低可能的检测限，2） 优化微流控测试芯片以进行最少的样品制备，3）算法开发和测试以获得 实时病毒检测与莫伊大学的合作伙伴一起评估可用性和利益相关者的需求 (Moi)和印第安纳州印第安纳波利斯的印第安纳州大学医学院（IUSM）将确保 手持平台和艾滋病毒检测将很容易被利益相关者采用。在R61结束时 在第一阶段，我们将有一个准备好进行实施测试的平台。 R33阶段的目标是开发一种概念验证设备，该设备在小型 生产运行，并根据血浆和全血中的临床性能指标进行全面评估。的 将对手持平台和测试芯片的过程控制再现性和初始试点临床试验进行评估。 研究将使用IUSM和Moy的HIV患者库存或前瞻性采集的样本进行 大学该R33阶段的结果将是高度表征的、敏感的、定量的手持式HIV。 病毒载量检测平台，其对真实的临床样本进行稳健的检测，并准备好进行规模放大。 完成这些目标将使我们能够快速获得FDA预提交和监管批准， 扩大规模，并在美国和东非实施高度准确的艾滋病毒载量自我检测。通过 使患者能够监测其HIV病毒载量，同时保持与医疗保健提供者支持的联系，我们 将有助于减少治疗依从性的障碍，并最终增加患者数量， 艾滋病规划署90-90-90目标所要求的持续病毒载量抑制。

SUMMARY ABSTRACT The 90-90-90 Target endorsed by UNAIDS, proposes diagnosing 90% of people infected with HIV worldwide, engaging 90% of them on effective antiretroviral treatment, and ensuring that 90% of those treated achieve sustained viral load suppression by 2020. The United States is woefully unprepared to reach this target; only 30% of HIV infected individuals in the U.S. currently achieve viral load suppression. Clearly, a paradigm shift in viral load testing will be required to accomplish this goal. This proposal aims to create a quantitative, handheld, viral load self-test that can be used by patients themselves to support their management of this chronic condition and that delivers relevant clinical insights to remote healthcare providers. Building on our preliminary work, the objective of the R61 phase is to combine isothermal loop- mediated nucleic acid amplification (LAMP) with a highly sensitive and label-free readout method called particle diffusometry for quantification of HIV viral load in a hand held platform with minimal process steps. Specific milestones include: 1) optimization of LAMP assays to ensure the lowest possible limit of detection, 2) optimization of a microfluidic test chip for minimal sample prep, 3) algorithm development and testing to obtain real-time viral detection. Assessment of usability and stakeholder needs with our partners at Moi University (Moi) in Eldoret, Kenya, and Indiana University School of Medicine (IUSM) in Indianapolis, Indiana will ensure that the handheld platform and HIV test will be readily adopted by stakeholders. At the conclusion of the R61 phase we will have a platform that is ready for implementation testing. The objective of the R33 phase is to develop a proof-of-concept device that is validated in small manufacturing runs and fully assessed against clinical performance metrics in plasma and whole blood. The handheld platform and test chips will be evaluated for process control reproducibility and an initial pilot clinical study will be performed with banked or prospectively collected samples from HIV patients at IUSM and Moy University. The outcome of this R33 Phase will be a highly characterized, sensitive, quantitative handheld HIV viral load detection platform that performs robustly with real clinical samples and is ready for scale up. Completion of these objectives will position us for FDA pre-submission and regulatory approval, rapid scale up, and implementation of a highly accurate HIV viral load self-test in the U.S. and East Africa. By enabling patients to monitor their HIV viral load while remaining connected to healthcare provider support, we will help to reduce barriers to treatment compliance and ultimately increase the number of patients with sustained viral load suppression as called for by the UNAIDS 90-90-90 targets.