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: 9204799
• 负责人: Gregory L Damhorst
• 金额: $ 4.9万
• 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/9204799
• 关键词: AIDS/HIV problem; Address; Affect; Antibodies; Area; 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; Heating; Immobilization; Individual; Infant; Infection; Ions; Lab-On-A-Chips; Laboratories; 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; microchip; nucleic acid detection; pandemic disease; particle; point of care; point-of-care diagnostics; portability; public health relevance; 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着世界上最贫穷的地区，因此，3400万生活在贫困地区的人中有很大一部分人 全世界感染该病毒的人都得不到足够的医疗照顾。这些人群获得适当治疗的一个主要障碍是缺乏用于监测疾病进展和评估抗逆转录病毒治疗疗效的诊断仪器。由于成本、尺寸和对熟练操作者的需要，标准仪器通常不实用，因此迫切需要低成本、用户友好的即时诊断测试。特别是，非常需要用于早期检测感染、确定接受抗逆转录病毒治疗的个体的血浆病毒载量以及测量艾滋病毒阳性母亲所生婴儿的CD4+ T淋巴细胞中的前病毒DNA的技术。 该项目旨在设计两个芯片实验室生物传感器平台，其将能够定量血浆中的HIV和受感染的CD4+ T细胞中的前病毒。这些平台将成为低成本、用户友好的即时检测设备的基础，该设备分析少量全血输入以确定HIV感染者的血浆病毒载量和前病毒DNA载量。虽然目前血浆病毒载量的金标准是核酸定量，但我们的方法是通过使用固定在微流体室表面上的病毒特异性抗体从全血样品中捕获血浆中的全病毒颗粒来检测血浆中的全病毒颗粒。冲洗掉血细胞和其他碎片后，捕获的病毒将用含有浓缩磷酸盐缓冲盐水（PBS）的脂质体标记。背景介质将通过低电导率介质冲洗掉，并且脂质体颗粒的泄漏将通过升高装置温度来触发，刺激离子的释放，其可以通过阻抗谱来量化。将根据阻抗变化确定病毒数量。 类似地，我们的前病毒检测方法是在微流体室中从全血样品中捕获CD4+ T淋巴细胞。随后用化学试剂裂解将细胞内内容物洗涤到反应室中，在那里它们可以被分析。通过PCR扩增病毒DNA（前病毒）或最终用电传感器检测，这消除了对光学组件的需要，并作为低成本和快速检测的优秀平台。 这些生物传感器技术将改变世界各地管理艾滋病毒的方式，使治疗更加知情，在偏远和资源有限的环境中更容易获得频繁的诊断数据。