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

项目摘要 NIH SBIR第一阶段提案的目标是开发一种低成本和快速的床旁（POC）p24检测， 早期发现人类免疫缺陷病毒（HIV）感染。它将适合在分散的 在发达国家和发展中国家，基础设施和实验室能力有限， 禁止病毒载量核酸检测。我们的目标是保持传统方法的准确性和灵敏度， 基于实验室的诊断，同时提供了一个设备，一个最低限度的培训人员可以操作，显著 增加获得艾滋病毒诊断的机会。世卫组织指南建议启动抗逆转录病毒治疗（ART）， 联合国艾滋病毒和艾滋病联合规划署（艾滋病规划署）呼吁， 全球艾滋病毒检测、治疗和病毒抑制的增长。然而，由于相当大的比例 的人不知道他们的感染，扩大检测能力有很大的潜力，以达到以前 未经检测的高危人群。目前可以直接检测HIV病毒血症的即时检测设备也 昂贵并且需要某种程度的专门培训来操作。此外，许多POC技术 目前已批准的新方法在病毒血症高峰期和急性感染的传播性中检测HIV的灵敏度不够。 现有的自我检测技术只能检测宿主抗体反应，这种反应是在最初的免疫反应后数周出现的。 感染因此，能够实现分散式测试的实用且经济实惠的POC测试平台将是 这对联邦“结束艾滋病流行：美国计划”倡议很重要。为此，AI Biosciences 提出开发一种低成本的基于磁性颗粒（MP）和纳米颗粒的夹心免疫测定法， 用于HIV-1 p24衣壳蛋白的早期检测，灵敏度接近核酸扩增 接近。这个小装置（~ 4x 3x 2英寸）执行基于MP的p24捕获和浓缩，然后 用抗体修饰的铂纳米颗粒（PtNPs）标记。然后PtNPs与氢原子相互作用， 过氧化氢溶液产生的信号，可以记录极高的灵敏度，使用低成本的电子 件.我们的目标是在第一阶段优化化学和测定形状因子，以制备30分钟的样品- 需要最少的培训和动手时间的测试。我们将与钟传健教授合作， 宾厄姆顿大学（BU）的纳米技术和催化专家， 抗体修饰的MP和PtNP。AI Biosciences还与乔治的Jeanne Jordan教授合作 华盛顿大学（GWU）协助进行器械评价和测试。在第一阶段结束时，我们将 我们的设备和分析到她的实验室进行技术演示。我们将培训技术人员使用 我们的设备和检测方法进行验证。他们将比较我们的免疫测定结果与PCR和第5代 代免疫测定。如果我们的敏感p24检测方法成功，我们将使用相同的方法。 检测HIV抗体的方法。

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.