Microchip for HBV testing using HIV-infected blood samples

使用感染艾滋病毒的血液样本进行乙型肝炎病毒检测的微芯片

• 批准号: 10538576
• 负责人: Hadi Shafiee
• 金额: $ 50.41万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10538576
• 关键词: Antibodies; Antigens; Biological Assay; Blood Volume; Blood specimen; Caring; Cellular Phone; Chronic; Cirrhosis; Complex; Cytomegalovirus; DNA; Detection; Development; Devices; Disease Management; Fingers; Fumarates; Gases; Goals; HIV; HIV Infections; Hepatitis; Hepatitis B Infection; Hepatitis B Virus; Hepatitis C; Herpesviridae; Human Herpesvirus 4; Human Herpesvirus 8; Hydrogen Peroxide; Individual; Infection; Influenza; Knowledge; Label; Laboratories; Link; Liver; Malaria; Methods; Microfluidic Microchips; Monitor; Morbidity - disease rate; Nanotechnology; Nucleic Acids; Optics; Patients; Performance; Persons; Platinum; Play; Point of Care Technology; Polymerase Chain Reaction; Primary carcinoma of the liver cells; Process; RNA; Regimen; Reverse Transcription; Risk; Role; Saliva; Sampling; Seminal fluid; Sensitivity and Specificity; Serology test; Services; Signal Transduction; Simplexvirus; Site; Specificity; System; Technology; Tenofovir; Testing; Treatment Efficacy; Treatment Failure; Treatment outcome; Urine; Venipunctures; Viral Load result; Virus; Virus Diseases; Whole Blood; Work; acute infection; clinically relevant; co-infection; cost; detection platform; diagnostic tool; disease diagnosis; empowerment; high risk; improved; infection risk; microchip; mortality risk; multiplex diagnostics; nanoparticle; nanoprobe; point of care; point of care testing; portability; rapid test; screening; technology platform; tool; transmission process

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.

项目摘要 据估计，超过2.4亿人患有HBV感染，超过3600万人 全世界感染HIV感染，其中高达60％的HBV感染和30％的HIV感染者是 不知道其感染状况。 HIV感染患者的HBV共同体与肝硬化的风险增加有关 和肝细胞癌（HCC），较高的慢性和隐匿性HBV率以及与实时相关的率较高 发病率。对HBV/HIV共感染状态的准确及时了解对于选择最佳艺术至关重要 方案和替诺福韦毒素富马酸酯（TDF）以及治疗监测以确定治疗开关是否 是必须的。先前的研究表明，使用现场POC测试对HIV和HBV的简单检测可以 显着改善筛查过程和与护理慢性病毒高风险的个人的联系 此外，未诊断，未经治疗的感染者可以在感染中发挥重要作用 向他人传播。如果未经治疗，感染可能会导致高级阶段，从而增加 死亡。缺乏适当的诊断工具，尤其是对于高感染风险的人来说是 感染不认识的原因。护理点（POC）多路复用诊断的发展是 对于患有HIV/HBV共同患者的肝炎测试和疾病管理服务至关重要 及时感染。病毒载荷测试是HIV和HBV的最准确和首选方法 检测和治疗监测。基于核酸的测定目前用于病毒载荷测试 疾病诊断和治疗效率监测，但是，这些测定仍然相对昂贵， 基于实验室，技术复杂。使用基于核酸的方法将HIV/HBV多路复用是 由于简单检测RNA（HIV）和DNA（HBV）靶标分子而挑战。此外， 当前的POC快速测试（量油）靶抗体/抗原感染后产生的HIV/HBV， 无法检测治疗失败和急性感染，并且具有低灵敏度/特异性。那就增加了 通过定期治疗监测获得HIV/HBV护理并改善治疗结果，有一个 迫切需要POC的廉价，快速，敏感和特定的HIV/HBV病毒负荷测试工具。这 这个高度跨学科项目的主要目标是开发便携式纳米技术授权 基于手机的系统（<30分钟）多路复用HIV/HBV的指纹体积（<100 µL）的整体系统 血液放在廉价（<$ 2），一次性和质量可实用的微流体装置上。在我们的建议中 方法，（i）使用高度特定的信封抗体在片上捕获靶病毒，（ii）捕获的病毒 用与相对抗体共轭的PT纳米颗粒（iii）标记的捕获病毒标记 在片上过氧化氢的情况下，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）的手机光学附件来量化气泡的定量/定性 病毒负荷测试。