Smartphone-based POC Testing for HIV Using Glowstick Chemistry

有效的艾滋病毒检测允许检测急性感染(新发和反弹)，适当 抗逆转录病毒治疗、抑制病毒载量和阻断传播。然而，早些时候 在需要时或在家里检测感染或定期自我监测仍然是一项挑战。侧向 流动分析是一种成熟、廉价的按需诊断方法，通常是 由于其易用性和低成本，在适用时受到青睐。他们最常跌倒的地方 简而言之，在分析灵敏度方面，尤其是在传染病的抗原检测诊断方面。我们的 目标是通过实施廉价、稳定的货架来从根本上改进LFA 发光棒的化学激发化学，用于在没有光或光学的情况下激发荧光 过滤器。我们的初步结果表明，这种化学物质使LFA的限制大大提高 检测(非常低pg/毫升)，使用标准智能手机，带有2美元的适配器，没有复杂的光学元件 组件。我们的中心假设是，我们的新的、试剂优化的化学激发侧向流动 与储存稳定的试剂盒集成的检测技术，可通过智能手机读取 将导致一种新的、广泛有用的、潜在的非常优越的高度敏感的侧向流动 化验。我们将通过5个具体目标来检验这一假设：(1)开发一种高度敏感和 简便易行的人血中HIV p24侧向流式细胞仪的研制 亲和力增强的双纤维连接蛋白结构域结合蛋白，(2)免疫复合物解离(ICD) 反弹/突破性艾滋病毒诊断，(3)对300名阴性和存档阳性的人进行检测 来自不同队列的样本，(4)使用患者的新鲜样本来评估临床表现 以及对艾滋病毒感染/反弹高危人群的可用性(R33阶段)，(5) 更多的人更容易使用，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).