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

基本信息

批准号：
10311486
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
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10311486
关键词：
Antibodies Antigens Biological Assay Blood Volume Blood specimen Caring Chronic Cirrhosis Complex Cytomegalovirus DNA Detection Development Devices Disease Management Fumarates Gases Goals Gold HIV HIV Infections Hepatitis Hepatitis B Infection Hepatitis B Virus Hepatitis C Human Herpesvirus 4 Human Herpesvirus 8 Hydrogen Peroxide Individual Infection Influenza Knowledge Label Laboratories Lead 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 Time Treatment Efficacy Treatment Failure Treatment outcome Urine Venipunctures Viral Load result Virus Virus Diseases Whole Blood Work acute infection base clinically relevant co-infection cost detection platform diagnostic tool disease diagnosis empowered high risk improved infection risk microchip mortality risk multiplex diagnostics nanoparticle nanoprobe point of care point of care testing portability rapid test screening tool transmission process

项目摘要

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.

项目概要据估计，有超过 2.4 亿人感染乙肝病毒，其中超过 3600 万人感染乙肝病毒。全球范围内都有艾滋病毒感染者，其中高达 60% 的 HBV 感染者和 30% 的 HIV 感染者不了解 HIV 感染者的 HBV 合并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 发生率较高、肝脏相关性发生率较高准确及时地了解 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二吡呋酯 (TDF) 以及治疗监测以确定治疗是否转换先前的研究表明，使用现场 POC 测试可以同时检测 HIV 和 HBV。显着改善筛查过程以及与慢性病毒高危人群护理的联系此外，未经诊断、未经治疗的感染者也可能在感染中发挥重要作用。如果不治疗，感染可能会发展到晚期，从而增加感染的风险。缺乏适当的诊断工具，特别是对于感染高危人群来说，是造成死亡率的原因之一。护理点 (POC) 多重诊断的发展是造成感染不知情的原因。对于扩大针对 HIV/HBV 感染者的肝炎检测和疾病管理服务至关重要及时检测感染情况，病毒载量检测是检测 HIV 和 HBV 最准确且首选的方法。基于核酸的检测目前用于病毒载量检测。疾病诊断和治疗效果监测，然而，这些检测方法仍然相对昂贵，使用基于核酸的方法对 HIV/HBV 进行多重检测也是基于实验室且技术复杂的。由于同时检测 RNA (HIV) 和 DNA (HBV) 目标分子，因此具有挑战性。目前的 POC 快速检测（试纸）针对的是感染后产生的 HIV/HBV 抗体/抗原，无法检测治疗失败和急性感染，并且敏感性/特异性较低，因此需要增加。通过定期治疗监测获得 HIV/HBV 护理并改善治疗结果，有一个迫切需要廉价、快速、灵敏且特异的 HIV/HBV 病毒载量即时检测工具。这个高度跨学科项目的主要目标是开发一种便携式纳米技术驱动的基于手机的系统，用于在指尖采血体积（<100 µL）中快速（<30 分钟）多重检测 HIV/HBV 在我们提出的方案中，血液被放置在廉价（<2 美元）、一次性且可大规模生产的微流体装置上。方法，(i) 使用高度特异性抗体在芯片上捕获包膜目标病毒，(ii) 捕获病毒用与各自抗体缀合的 Pt 纳米颗粒进行标记，(iii) Pt 标记的捕获病毒在芯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）手机光学附件对气泡进行量化，以进行定量/定性病毒载量测试。