Microchip virus detection platforms for point-of-care HIV/AIDS diagnostics

用于即时艾滋病毒/艾滋病诊断的微芯片病毒检测平台

• 批准号: 8997059
• 负责人: Gregory L Damhorst
• 金额: $ 4.86万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-12 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8997059
• 关键词: AIDS/HIV problem; Address; Affect; Antibodies; Area; Binding; Binding Proteins; Biological Assay; Biosensor; Blood; Blood Cells; Blood specimen; CD4 Positive T Lymphocytes; Caring; Cell Line; Cells; Chemicals; Clinical; Cytolysis; DNA; Data; Detection; Development; Devices; Diagnostic; Diagnostic tests; Disease Progression; Dyes; Early Diagnosis; Electrodes; Encapsulated; Enzyme-Linked Immunosorbent Assay; Enzymes; Fluorescence; Future; Generations; Gold; HIV; HIV Envelope Protein gp120; HIV Genome; HIV Infections; HIV Seropositivity; Health; Heating; Individual; Infant; Infection; Ions; Laboratories; Life; Lipid Bilayers; Lipids; Liposomes; Lymphocyte; Maternal antibody; Measurement; Medical; Microfluidic Microchips; Microfluidics; Monitor; Mothers; Nucleic Acid Amplification Tests; Nucleic Acids; Optics; Pattern; Phosphate Buffer; Plasma; Platinum; Population; Protocols documentation; Proviruses; Reaction; Reagent; Reporter; Resources; Risk; Saline; Series; Serologic tests; Spectrum Analysis; Surface; Techniques; Technology; Temperature; Testing; Therapeutic Intervention; Time; Transition Temperature; Viral; Viral Load result; Virion; Virus; Whole Blood; Work; antiretroviral therapy; base; cost; design; electric impedance; instrumentation; micro-total analysis system; microchip; nucleic acid detection; pandemic disease; particle; point of care; point-of-care diagnostics; portability; rapid detection; receptor; sensor; seroconversion; user-friendly; viral DNA; viral detection

DESCRIPTION (provided by applicant): The HIV pandemic disproportionately affects the poorest regions of the world and, as a result, a large portion of the 34 million individuals living with the virus worldwide do not have access to adequate medical care. One major barrier to proper treatment in these populations is the unavailability of diagnostic instrumentation for monitoring disease progression and assessing the efficacy of antiretroviral therapy. Standard instrumentation is often not practical because of cost, size and the need for a skilled operator, thus low-cost, user-friendly point-of-care diagnostic tests are desperately needed. In particular, technologies are greatly needed for early detection of infection, determination of plasma viral load in individuals undergoing antiretroviral therapy, and measurement of proviral DNA in CD4+ T lymphocytes in infants born to an HIV-positive mother. This project aims to design two lab-on-a-chip biosensor platforms which will be capable of quantification of HIV in blood plasma and provirus in infected CD4+ T cells. These platforms will be the basis for low-cost, user- friendly point-of-care detection devices which analyze a small volume of whole blood input to determine plasma viral load and proviral DNA load for individuals living with HIV. While the current gold standard for plasma viral load is nucleic acid quantification, our approach is to sense whole virus particles in blood plasma by capturing them from a whole blood sample using virus-specific antibodies immobilized on the surface of a microfluidic chamber. After rinsing away blood cells and other debris, the captured viruses will be tagged with liposomes containing concentrated phosphate buffered saline (PBS). The background media will be rinsed away by low conductivity media and leaking of the liposome particles will be triggered by raising the device temperature, stimulating the released of ions which can be quantified by impedance spectroscopy. Virus number will be determined from the change in impedance. Similarly, our approach to provirus detection is to capture CD4+ T lymphocytes in a microfluidic chamber from a whole blood sample. Subsequent lysis with a chemical reagent will wash intracellular contents into a reaction chamber where they can be analyzed. Amplification of viral DNA (provirus) by PCR or will ultimately be detected with electrical sensors which eliminate the need for optical components and serves as an excellent platform for low-cost and rapid detection. These biosensor technologies will change the way HIV is managed around the world, making treatment more informed and frequent diagnostic data more accessible in remote and resource-limited settings.

描述（由申请人提供）：艾滋病毒大流行不成比例地影响世界上最贫穷的地区，因此，居住的3400万个人中有很大一部分 在全球范围内，该病毒无法获得足够的医疗服务。在这些人群中，适当治疗的一个主要障碍是无法在监测疾病进展和评估抗逆转录病毒疗法的疗效的诊断仪器上可用。由于成本，大小和对熟练的操作员的需求，标准仪器通常不实用，因此迫切需要低成本，用户友好的护理诊断测试。特别是，对于正在接受抗逆转录病毒疗法的个体中血浆病毒载量的早期检测，确定血浆病毒载量以及测量患者HIV阳性母亲的儿童CD4+ T淋巴细胞的测量非常需要技术。 该项目旨在设计两个实验室芯片生物传感器平台，这些平台将能够在感染的CD4+ T细胞中量化血浆中的HIV和病毒病。这些平台将是低成本，用户友好的护理点检测设备的基础，该设备分析了少量全血输入，以确定HIV患者的血浆病毒载荷和前病毒DNA负载。尽管当前的血浆病毒载荷的金标准是核酸定量，但我们的方法是通过使用固定在微富集室表面上的病毒特异性抗体从全血样品中捕获血浆中的整个病毒颗粒。冲走血细胞和其他碎屑后，捕获的病毒将用含有浓磷酸盐缓冲盐水（PBS）的脂质体标记。背景培养基将被低电导率介质冲洗掉，脂质体颗粒的泄漏将通过提高设备温度触发，刺激离子释放的释放，这可以通过阻抗光谱法量化。病毒编号将从阻抗的变化中确定。 同样，我们的病毒检测方法是从全血样品中捕获微流体腔中的CD4+ T淋巴细胞。随后用化学试剂裂解将将细胞内含量洗净到可以分析的反应室中。通过PCR扩增病毒DNA（Provirus），或最终将用电气传感器检测到，从而消除了对光学组件的需求，并作为低成本和快速检测的绝佳平台。 这些生物传感器技术将改变世界各地的艾滋病毒管理方式，使治疗在远程和资源有限的设置中更容易获得知情和频繁的诊断数据。