Smartphone-based POC Testing for HIV Using Glowstick Chemistry

对艾滋病毒的有效检测可以发现急性感染（新发和反弹）， 抗逆转录病毒治疗、抑制病毒载量和阻断传播。然而，早 在需要时或在家中检测感染或定期自我监测仍然是一个挑战。横向 流动测定是一种完善的、廉价的即时诊断方法， 由于其易于使用和低成本，在适用时受到青睐。它们最常掉落的地方 不足之处在于分析灵敏度，特别是在传染病的抗原检测诊断方面。我们 我们的目标是通过实施廉价、货架稳定的 化学激发化学发光棒在一个点的护理格式激发荧光没有光或光学 filters.我们的初步结果表明，这种化学物质使LFA具有大大改善的限制， 检测（极低pg/mL），使用带有2美元适配器的标准智能手机， 件.我们的中心假设是，我们的新的，试剂优化的化学激发侧流 与储存稳定的试剂盒集成并可通过智能手机读取的分析技术 将导致一个新的，广泛有用的和潜在的非常优越的高度敏感的横向流类 测定。我们将通过5个具体目标来检验这一假设：（1）开发高度敏感且 用于人血液中HIV p24的易于使用的侧流测定，包括开发新的 亲和力增强的双纤连蛋白结构域结合剂，（2）免疫复合物解离（ICD）， 反弹/突破性艾滋病毒诊断，（3）对300名阴性和存档阳性人进行检测 来自不同队列的样本，（4）使用来自患者的新鲜样本评价临床性能 和HIV感染/反弹高风险的不同受试者人群的可用性（R33阶段），（5） 让更多人使用更好的可用性，测试项目的质量保证（R33阶段）。

Effective testing for HIV allows for detection of acute infections (new and rebound), appropriate antiretroviral treatment, suppression of viral load, and interruption of transmission. However, early detection of infection or regular self-monitoring at point-of-need or at home remains a challenge. Lateral flow assays are a well-established, inexpensive point-of-need diagnostic method, and commonly are favored when applicable because of their ease of use and low cost. Where they most commonly fall short is in analytical sensitivity, notably in antigen-detection diagnostics for infectious diseases. Our goal is to develop radically-improved LFAs by implementing the inexpensive, shelf-stable chemi-excitation chemistry of glow sticks in a point-of-care format to excite fluors without light or optical filters. Our preliminary results suggest this chemistry enables LFAs with greatly improved limits of detection (very low pg/mL), using a standard smartphone with a $2 adaptor and no sophisticated optical components. Our central hypothesis is that our novel, reagent-optimized chemi-excited lateral flow assay technology integrated with a storage-stable reagent cartridge and readable with a smartphone will lead to a new, widely-useful and potentially very-superior class of highly-sensitive lateral flow assays. We will test this hypothesis through 5 specific aims: (1) Development of a highly-sensitive and easily-usable lateral flow assay for HIV p24 in human blood, including development of novel avidity-enhanced dual-fibronectin-domain binders, (2) Immune complex dissociation (ICD) for rebound/breakthrough HIV diagnosis, (3) Testing with 300 negative and archived-positive human samples from diverse cohorts, (4) To evaluate clinical performance using fresh samples from patients and usability with a diverse population of subjects at high risk of HIV infection/rebound (R33 Phase), (5) Easier usability by more people, quality assurance for test items (R33 Phase).