Label-free HIV-1 Viral Load Measurement through a Contactless Electrode Microchip

通过非接触式电极微芯片进行无标记 HIV-1 病毒载量测量

• 批准号: 8527386
• 负责人: Hadi Shafiee
• 金额: $ 5.39万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8527386
• 关键词: Acoustics; Acquired Immunodeficiency Syndrome; Acute; Africa South of the Sahara; Anti-Retroviral Agents; Antibodies; Biological; Biological Assay; Blood specimen; Breast; Care Technology Points; Caring; Cells; Cessation of life; Child; Clinical; Communicable Diseases; Complex; Cytolysis; Detection; Detergents; Developing Countries; Devices; Diagnosis; Diagnostic; Disease; Doctor of Philosophy; Early Diagnosis; Electrodes; Enzyme Immunoassay; Enzyme-Linked Immunosorbent Assay; Goals; Gold; HIV; HIV Envelope Protein gp120; HIV-1; Human immunodeficiency virus test; Individual; Infection; Investigation; Label; Laboratories; Lateral; Left; Life; Malaria; Medicare; Medicine; Methods; Microelectrodes; Microfluidic Microchips; Microfluidics; Modeling; Monitor; Nanotechnology; Nucleic Acid Amplification Tests; Patients; Pattern; Persons; Pharmaceutical Preparations; Plasma; Play; Population; Public Health; Reporting; Resources; Role; Sampling; Security; Sensitivity and Specificity; Staging; Streptavidin; System; Technology; Testing; Tuberculosis; Vertical Disease Transmission; Viral; Viral Load result; Viral load measurement; Virus; Virus Diseases; Whole Blood; base; clinical Diagnosis; cost; disease diagnosis; electric field; electric impedance; fluorescence imaging; interest; leukemia; magnetic beads; medical schools; microchip; mortality; nano; nanomagnetic; novel; point of care; prostate cancer cell; prototype; public health relevance; screening; statistics; tool; transmission process

DESCRIPTION: Rapid, inexpensive, and early detection of infectious viral diseases is an urgent unmet need with diverse applications ranging from clinical diagnosis, public health, and homeland security. Among these applications, Human Immunodeficiency Virus (HIV-1) diagnostics in resource-limited settings plays a critical role to provide appropriate and timely care to patients. More than 95% of deaths due to infectious diseases such as malaria, Acquired Immune Deficiency Syndrome (AIDS), and tuberculosis (TB) have been reported to occur in developing countries. A significant ratio (67%) of the 33.3 million HIV-1 infected population live in Sub-Saharan Africa. These statistics clearly highlight the urgent demand for rapid, inexpensive, and simple screening tests to identify infected individuals for HIV/AIDS treatment. However, current HIV-1 diagnostics such as lateral flow assays, including dipsticks or enzyme immunoassays (ELISA), and OraQuick HIV test kit lack the capability to detect acute HIV-1 infection even at high HIV-1 viral load. Detecting persons with acute HIV-1 infection is crucial since viral replication and shedding occur in this stage before detectable HIV-1 antibodies appear. Persons with acute HIV-1 infection are unaware of their disease and therefore contribute substantially to HIV-1 transmission. At this stage, there is maximum viral replication and shedding (108 copies/mL). Thus, there is an immediate need for easy to use, portable, and inexpensive diagnostic tool for acute HIV-1 detection (seroconversion and asymptomatic stages) at the POC. At the Bio-Acoustic MEMS in Medicine laboratory at Harvard Medical School, Dr. Demirci's (Sponsor) team has presented prototype viral load counting platforms based on lensless and fluorescence imaging. Here, I will employ my previous expertise in microfluidic cell electro-manipulation and my current expertise in antibody based HIV-1 virus capturing using micro/nano technologies to create a POC HIV-1 viral load measurement platform for treatment initiation and monitoring. Streptavidin-coated nano-magnetic beads conjugated with gp120 antibody will be used to selectively capture and isolate HIV-1 viruses from whole blood samples. The nanobead-HIV virus complexes will be then introduced into the microfluidic device with the optimized contactless microelectrodes functionalized with gp120 antibody to capture the nanobead-HIV complex. The captured viruses will be lysed through introducing a lysis detergent and the impedance change will be correlated to the viral load of the sample. This proposal will provide an inexpensive method for HIV-1 viral load measurements at POC that is compatible with mass fabrication technologies with a great promise to be commercialized.

描述：快速，廉价且早期发现传染病病毒疾病是紧迫的未满足需求，从临床诊断，公共卫生和国土安全性等不同的应用中。在这些应用中，在资源有限的设置中的人类免疫缺陷病毒（HIV-1）诊断起着至关重要的作用，可以为患者提供适当和及时的护理。据报道，由于传染病，疟疾，获得的免疫缺陷综合征（AIDS）和结核病（TB）等导致的死亡人数超过95％，在发展中国家发生。在撒哈拉以南非洲，3330万HIV-1感染人群中有很大的比例（67％）。这些统计数据显然强调了对快速，廉价和简单的筛查测试的紧迫需求，以识别受感染的艾滋病毒/艾滋病治疗的人。然而，当前的HIV-1诊断（例如侧向流量测定），包括量子或酶免疫测定（ELISA）和ORAQUICK HIV测试套件，即使在高HIV-1病毒载量下，也缺乏检测急性HIV-1感染的能力。检测患有急性HIV-1感染的人至关重要，因为在可检测到的HIV-1抗体出现之前，病毒复制和脱落发生在此阶段。急性HIV-1感染的人没有意识到自己的疾病，因此对HIV-1传播产生了重大贡献。在此阶段，有最大的病毒复制和脱落（108份/ml）。因此，在POC处易于使用易于使用，便携式和廉价的诊断工具（血清转换和无症状阶段）。在哈佛医学院的医学实验室的生物声学磁场上，Demirci博士（赞助商）团队根据无透镜和荧光成像提出了原型病毒负载计数平台。在这里，我将采用以前的微流体电池电化学专业知识，以及我当前在抗体基于HIV-1病毒方面的专业知识，使用微/纳米技术捕获，以创建POC HIV-1病毒负荷测量平台，以进行治疗起始和监测。与GP120抗体结合的链霉亲和素涂层的纳米磁珠将用于选择性捕获和分离全血样品中的HIV-1病毒。然后，将使用优化的非接触式微电极与GP120抗体功能化，以捕获纳米果-EV-HIV复合物，然后将纳米果-HIV病毒复合物引入微流体设备。捕获的病毒将通过引入裂解洗涤剂来裂解，阻抗变化将与样品的病毒负荷相关。该提案将为POC的HIV-1病毒负荷测量提供廉价的方法，该方法与大众制造技术兼容，并有很大的承诺被商业化。