Highly Sensitive HIV Viral Load Assay for Point-of-Care Settings

项目概要： 据世界卫生组织（世卫组织）报告，目前有3670多万人感染艾滋病 最近到目前为止，艾滋病毒已造成3500多万人死亡。疫情持续增长，近2.1 2015年新增艾滋病毒感染病例100万例，其中美国新增艾滋病毒感染病例超过4.7万例。目前， 在发展中国家，只有34%的艾滋病毒感染者接受抗逆转录病毒疗法（ART）治疗 尽管抗逆转录病毒治疗是负担得起的，甚至是免费提供的。减少艾滋病毒的根本挑战 负担及其流行包括农村地区缺乏对病毒载量的即时检测， 获得基本实验室基础设施和训练有素的技术人员的机会有限。在发达国家，HIV-1 病毒载量定期用于密切监测和评估患者对ART的反应，以确保药物 坚持和分期疾病进展。相比之下，发展中国家正在使用CD 4 + T细胞， 淋巴细胞计数和临床症状，以指导ART遵循WHO指南，但 需要进行病毒载量检测的婴儿。然而，研究表明，CD 4 + T细胞 计数策略不能检测早期病毒学失败，导致耐药病毒积累 感染个体中的菌株，并降低ART疗效。因此，提供了一种快速的、廉价的、一次性的且 迫切需要简单的病毒载量测试。目前的病毒载量技术是基于定量 HIV-1 RNA、p24抗原和逆转录酶活性。这些HIV-1病毒载量测定是 昂贵（每次测试50 -200美元），技术复杂，因此需要训练有素的技术人员运行 测定。在这里，我们将利用我们在微芯片制造，微流体，表面功能化， 无透镜成像和生物传感，以创建一次性HIV-1病毒载量芯片， 从全血/血浆中捕获HIV，（ii）高度敏感（可检测<1000个病毒拷贝/mL）， 便宜（约1美元），快速（45分钟内），（iii）可处理少量血浆/全血 (1mL)以及（iv）高度稳定（无需冷藏）以帮助POC的HIV定量和治疗 设置.我们设想，所提出的技术将有助于快速病毒载量定量， 全世界数百万的患者。 !

PROJECT SUMMARY: More than 36.7 million people are living with AIDS as reported by World Health Organization (WHO) recently. HIV caused over 35 million deaths so far. The epidemic continues to grow with nearly 2.1 million new cases of HIV in 2015 including over 47,000 new cases of HIV infection in US. Currently, only 34% of HIV infected people are getting antiretroviral therapy (ART) treatment in developing world despite ART being affordable or even freely available. The fundamental challenges to reduce HIV burden and its prevalence include the lack of point-of-care assays for viral load in rural areas having limited access to basic laboratory infrastructure and trained technicians. In developed countries, HIV-1 viral load is regularly used to closely monitor and assess the patient response to ART, to ensure drug adherence and to stage disease progression. In contrast, developing countries are using CD4+ T lymphocyte count and clinical symptoms to guide ART following the WHO guidelines with the exception of infants, where viral load assays are required. However, studies have shown that CD4+ T cell counting strategy cannot detect early virological failure which leads to accumulation of drug-resistant strains in infected individuals and reduces ART efficacy. Thus, a rapid, inexpensive, disposable, and simple viral load test is urgently needed. Current viral load technologies are based on the quantification of HIV-1 RNA, p24 antigen, and reverse transcriptase activity. These HIV-1 viral load assays are expensive ($50-200 per test), and technically complex, thus requiring trained technicians to run the assays. Here, we will use our expertise in microchip fabrication, microfluidics, surface functionalization, lensless imaging, and biosensing to create a disposable HIV-1 viral load chip that can (i) selectively capture HIV from whole blood/plasma, (ii) be highly sensitive (can detect <1000 viral copies/mL), inexpensive (~$1), and rapid (within 45 minutes), (iii) can handle small volume of plasma/whole blood (1mL) and (iv) be highly stable (refrigeration free) to aid in HIV quantification and treatment at POC settings. We envision that the proposed technology will aid in rapid viral load quantification benefiting millions of patients around the world. !