An Ultrasensitive and Low-Cost p24 Antigen Test for the Early Detection of HIV

项目总结 NIH SBIR第一阶段提案的目标是开发一种低成本、快速的护理点(POC)p24测试，用于 及早发现人类免疫缺陷病毒(HIV)感染。它将适合在分散的环境中使用 发达国家和发展中国家的环境，那里的基础设施和实验室能力有限 禁止基于病毒载量的核酸检测。我们的目标是保持传统的准确性和敏感度 基于实验室的诊断，同时提供受过最低程度培训的人员都能操作的设备，意义重大 增加获得艾滋病毒诊断的机会。世卫组织指南建议开始抗逆转录病毒治疗(ART) 感染后越早越好，联合国艾滋病毒和艾滋病联合规划署(艾滋病规划署)呼吁 用于全球艾滋病毒检测、治疗和病毒抑制的增加。然而，因为有相当大的比例 的人没有意识到自己的感染，扩大检测能力很有可能在以前达到 未经测试的高危人群。目前可以直接检测艾滋病毒病毒血症的护理点设备也是 价格昂贵，需要一定程度的专业培训才能操作。此外，许多PoC技术具有 已经批准的方法不够敏感，不足以在病毒血症高峰期检测艾滋病毒，也不能在急性感染中检测到传播性。 现有的自我检测技术只检测宿主抗体反应，这种反应在最初的几周后出现 感染。因此，支持分散测试的实用且负担得起的PoC测试平台将是 对联邦“结束艾滋病毒流行：美国的计划”具有重要意义。为此，AI生物科学 建议开发一种低成本的磁粒子(MP)和纳米粒子为基础的夹心免疫分析，可以 用于HIV-1p24衣壳蛋白的早期检测，灵敏度接近核酸扩增 接近了。这个小型设备(约4x3x2英寸)执行基于MP的p24捕获和集中，然后 抗体修饰的铂纳米颗粒(PtNPs)标记。然后，PtNPs与氢气相互作用 过氧化氢溶液，可以用低成本的电子设备以极高的灵敏度记录信号 组件。我们的目标是在第一阶段优化化学和检测形状因素，以制作30分钟的样品- 需要最少的培训和实际操作时间的答题测试。我们将与钟传健教授合作， 宾厄姆顿大学(BU)纳米技术和催化专家，表征和合成 抗体修饰的MPS和PtNPs。AI Biosciences还与乔治大学的珍妮·乔丹教授合作 华盛顿大学(GWU)协助进行设备评估和测试。在第一阶段结束时，我们将带来 我们的仪器和化验设备送到她的实验室进行技术演示。我们将培训技术人员使用 我们的设备和化验以供验证。他们会将我们的免疫检测结果与pcr和5号进行比较。 世代免疫分析。如果我们的方法对敏感的p24检测成功，我们将使用相同的 艾滋病病毒抗体检测方法。

PROJECT SUMMARY The goal of this NIH SBIR Phase I proposal is to develop a low-cost and rapid point-of-care (POC) p24 test for early detection of human immunodeficiency virus (HIV) infection. It will be appropriate for use at decentralized settings in developed and developing countries where the limitations of infrastructure and laboratory capability prohibit viral load-based nucleic acid testing. Our aim is to maintain the accuracy and sensitivity of traditional lab-based diagnostics while providing a device that a minimally trained person can operate, significantly increasing access to HIV diagnostics. The WHO guidelines recommend initiating antiretroviral therapy (ART) as early as possible once infected, and the Joint United Nations Programme on HIV and AIDS (UNAIDS) has called for global increases in HIV testing, treatment, and viral suppression. However, because a significant proportion of individuals are unaware of their infection, expanding testing capabilities has a high potential to reach previously untested high-risk populations. Current point-of-care devices that can detect HIV viremia directly are too expensive and require some level of specialized training to operate. Also, many POC technologies that have been approved are not sensitive enough to detect HIV during peak viremia and transmissibility in acute infection. Existing self-testing technologies only detect the host antibody response, which arises weeks after the initial infection. Therefore, practical and affordable POC test platforms that enable decentralized testing will be important for the federal “Ending the HIV Epidemic: A Plan for America” initiative. To this end, AI Biosciences proposes to develop a low-cost magnetic particle (MP) and nanoparticle based sandwich immunoassay that can be used for the early-detection of HIV-1 p24 capsid protein with a sensitivity near that of nucleic acid amplification approaches. This small device (~4x3x2 inches) performs MP-based p24 capture and concentration, followed by labeling with antibody-modified platinum nanoparticles (PtNPs). The PtNPs then interact with a hydrogen peroxide solution to create a signal that can be recorded with extremely high sensitivity using low-cost electronic components. We aim to optimize the chemistry and assay form factor during Phase I to make a 30-min sample- to-answer test that requires minimal training and hands-on time. We will work with Professor Chuan-Jian Zhong, a nanotechnology and catalysis expert at Binghamton University (BU), to characterize and synthesize the antibody modified MPs and PtNPs. AI Biosciences has also partnered with Professor Jeanne Jordan of George Washington University (GWU)to assist with device evaluation and testing. At the end of Phase I, we will bring our device and assay to her laboratory to perform a technology demonstration. We will train technicians to use our device and assay for validation. They will compare the results from our immunoassay to PCR and 5th generation immunoassays. If our approach for sensitive p24 detection is successful, we will use the same approach for HIV antibody detection.