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、廉价和早期检测传染性病毒性疾病是临床诊断、公共卫生和国土安全等多种应用中未满足的迫切需求。在这些应用中，资源有限环境中的人类免疫缺陷病毒 (HIV-1) 诊断在为患者提供适当和及时的护理方面发挥着关键作用。据报道，95%以上因疟疾、获得性免疫缺陷综合症（艾滋病）和结核病（结核病）等传染病造成的死亡发生在发展中国家。 3330 万 HIV-1 感染者中很大一部分 (67%) 生活在撒哈拉以南非洲地区。这些统计数据清楚地突出了对快速、廉价和简单的筛查测试的迫切需求，以识别感染者并进行艾滋病毒/艾滋病治疗。然而，当前的 HIV-1 诊断方法，例如侧向层析检测，包括试纸或酶免疫分析 (ELISA) 和 OraQuick HIV 检测试剂盒，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。检测急性 HIV-1 感染者至关重要，因为病毒复制和脱落发生在可检测到的 HIV-1 抗体出现之前的这一阶段。急性 HIV-1 感染者并不知道自己患病，因此对 HIV-1 传播起到了很大的推动作用。在此阶段，病毒复制和脱落达到最大（108 拷贝/mL）。因此，迫切需要易于使用、便携式且廉价的诊断工具，用于 POC 急性 HIV-1 检测（血清转化和无症状阶段）。在哈佛医学院的生物声学 MEMS 医学实验室，Demirci 博士（赞助商）团队展示了基于无透镜和荧光成像的原型病毒载量计数平台。在这里，我将利用我之前在微流体细胞电操作方面的专业知识和我目前在使用微/纳米技术捕获基于抗体的 HIV-1 病毒方面的专业知识，创建一个用于治疗启动和监测的 POC HIV-1 病毒载量测量平台。与 gp120 抗体缀合的链霉亲和素包被的纳米磁珠将用于从全血样本中选择性捕获和分离 HIV-1 病毒。然后将纳米珠-HIV 病毒复合物引入微流体装置中，并使用经过 gp120 抗体功能化的优化非接触式微电极来捕获纳米珠-HIV 复合物。捕获的病毒将通过引入裂解去污剂来裂解，并且阻抗变化将与样品的病毒载量相关。该提案将为 POC 测量 HIV-1 病毒载量提供一种廉价的方法，该方法与大规模制造技术兼容，有望实现商业化。