Smartphone-based diagnostic for HIV self-testing

摘要摘要 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， 让其中90%的人接受有效的抗逆转录病毒治疗，并确保90%的接受治疗的人实现 到2020年，病毒载量持续受到抑制。可悲的是，美国对实现这一目标毫无准备；只有 在美国，30%的艾滋病毒感染者目前实现了病毒载量抑制。显然，中国的范式转变 为了实现这一目标，将需要进行病毒载量测试。这项提议旨在创造一种量化的、手持的、 可由患者自己使用的病毒负载自我测试，以支持他们对此慢性疾病的管理 并向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阶段的目标是将等温回路- 一种高灵敏度、无标记的读出方法 用最少的过程步骤在手持平台中定量艾滋病毒病毒载量的粒子扩散计量法。 具体的里程碑包括：1)优化LAMP分析，以确保尽可能低的检测下限，2) 最小样品制备微流控芯片的优化，3)算法开发和测试 实时病毒检测。与我们在莫伊大学的合作伙伴一起评估可用性和利益相关者需求 肯尼亚埃尔多雷特的MOI和印第安纳州印第安纳波利斯的印第安纳大学医学院(IUSM)将确保 手持平台和艾滋病毒检测将很容易被利益攸关方采用。在R61的结束时 阶段我们将拥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在小范围内得到验证的概念验证设备 生产运行，并根据血浆和全血的临床表现指标进行全面评估。这个 将评估手持平台和测试芯片的过程控制重复性和初步临床试验 研究将使用IUSM和MOY储存的或预期收集的艾滋病毒患者的样本进行 大学。R33阶段的结果将是一种高度特征化的、敏感的、定量的手持艾滋病毒 病毒载量检测平台，在实际临床样本中表现强劲，并已准备好扩大规模。 完成这些目标将使我们能够快速获得FDA的预提交和监管部门的批准 扩大规模，并在美国和东非实施高精度的艾滋病毒病毒载量自我检测。通过 使患者能够监控他们的HIV病毒载量，同时保持与医疗保健提供商的支持联系，我们 将有助于减少治疗依从性的障碍，并最终增加患有 按照联合国艾滋病规划署90-90-90目标的要求，持续抑制病毒载量。

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.