Microchip for HBV testing using HIV-infected blood samples

项目摘要 据估计，超过2.4亿人患有HBV感染，超过3600万人 全球范围内的HIV感染者，其中高达60%的HBV感染者和30%的HIV感染者 不知道自己的感染状况。HIV感染者合并HBV感染与肝硬化风险增加有关 和肝细胞癌（HCC），慢性和隐匿性HBV的发生率较高， 发病率。准确及时地了解HBV/HIV合并感染状况对于选择最佳ART至关重要 方案和富马酸替诺福韦酯（TDF）以及治疗监测，以确定是否转换治疗 是必需的.以前的研究表明，同时检测艾滋病毒和乙型肝炎病毒使用现场POC测试， 显著改善筛查过程，并与慢性病毒感染高危人群的护理联系 感染.此外，未确诊、未经治疗的感染者也可能在感染中发挥重要作用 传递给别人。如果不治疗，感染可能导致晚期，这增加了 mortality.缺乏适当的诊断工具，特别是对感染风险高的人来说， 感染原因不明。床旁（POC）多路复用诊断的发展是 对于扩大HIV/HBV感染者的肝炎检测和疾病管理服务至关重要， 及时感染。病毒载量检测是HIV和HBV最准确和首选的方法 检测和治疗监测。基于核酸的测定目前用于病毒载量检测， 然而，疾病诊断和治疗功效监测，这些测定仍然相对昂贵， 实验室为基础，技术复杂。使用基于核酸的方法多重HIV/HBV也是 由于同时检测RNA（HIV）和DNA（HBV）靶分子，因此具有挑战性。此外，委员会认为， 目前的POC快速测试（试纸）针对感染后产生的抗HIV/HBV的抗体/抗原， 不能检测治疗失败和急性感染，敏感性/特异性低。因此，为了增加 获得艾滋病毒/乙型肝炎护理与定期治疗监测和改善治疗结果，有一个 POC迫切需要廉价、快速、灵敏和特异的HIV/HBV病毒载量检测工具。的 这个高度跨学科的项目的主要目标是开发一个便携式纳米技术授权 一种基于手机的系统，用于快速（<30分钟）多重HIV/HBV的手指采血量（<100 µL）， 将血液置于廉价（<2美元）、一次性和可批量生产的微流体装置上。在我们提出的 方法，（i）使用高度特异性包膜抗体在芯片上捕获靶病毒，（ii）捕获的病毒 用与相应抗体缀合的Pt纳米颗粒标记，（iii）Pt标记的捕获病毒 在片上存在过氧化氢的情况下通过Pt纳米颗粒的气体形成产生气泡，（iv） 可以使用低成本（<4美元）的手机光学附件来量化气泡， 病毒载量检测

PROJECT SUMMARY It is estimated that more than 240 million people are living with HBV infection and more than 36 million people with HIV infection worldwide, of which up to 60% of HBV-infected and 30% of HIV-infected individuals are unaware of their infection status. HBV coinfcetion in HIV-infected patients is linked to increased risk of cirrhosis and Hepatocellular carcinoma (HCC), higher rate of chronicity and occult HBV, and higher rate of liver-related morbidity. Accurate and timely knowledge of HBV/HIV coinfection status is essential to select the optimal ART regimen and tenofovir disoproxil fumarate (TDF) as well as treatment monitoring to identify if treatment switch is required. Previous studies showed that simultaneous detection of HIV and HBV using on-site POC tests can significantly improve the screening process and linkage to care for individuals at high risk for chronic viral infections. In addition, the undiagnosed, untreated, infected individuals can play a significant role in infection transmission to others. If untreated, the infection can lead to advanced stages, which increases the risk of mortality. The lack of appropriate diagnostic tools especially for people at high risk of infection is one of the reasons for the infection unawareness. The development of point-of-care (POC) multiplexed diagnostics is crucial in expanding hepatitis testing and disease management services for individuals with HIV/HBV co- infection in a timely manner. Viral load testing is the most accurate and preferred approach for HIV and HBV detection and treatment monitoring. Nucleic acid-based assays are currently used for viral load testing for disease diagnosis and treatment efficacy monitoring, however, these assays are still relatively expensive, laboratory-based, and technically complex. Multiplexing HIV/HBV using nucleic acid-based methods is also challenging due to simultaneous detection of RNA (HIV) and DNA (HBV) target molecules. Furthermore, current POC rapid tests (dipsticks) target antibodies/antigens against HIV/HBV generated after infection, cannot detect treatment failure and acute infection, and have low sensitivity/specificity. Thus, to increase access to HIV/HBV care with regular treatment monitoring and to improve treatment outcomes, there is an urgent need for inexpensive, rapid, sensitive, and specific HIV/HBV viral load testing tools at the POC. The main goal of this highly interdisciplinary project is developing a portable nanotechnology-empowered cellphone-based system for rapid (<30 minutes) multiplexing HIV/HBV in fingerprick volume (<100 µL) of whole blood placed on an inexpensive (<$2), disposable, and mass-producible microfluidic device. In our proposed method, (i) target viruses are captured on-chip using highly specific envelope antibodies, (ii) captured viruses are labeled with Pt nanoparticles conjugated with respective antibodies, (iii) Pt-labeled captured viruses generate bubbles through gas formation of Pt nanoparticles in the presence of hydrogen peroxide on-chip, (iv) the bubbles can be quantified using a low-cost (<$4) cellphone optical attachment for quantitative/qualitative viral load testing.