LLISA: ???Liposome-Linked Immunosorbant Assay??? for Detection of HIV Viral Load

LLISA：???脂质体连接免疫吸附测定???

• 批准号: 8514874
• 负责人: Rashid Bashir
• 金额: $ 18.12万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-15 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8514874
• 关键词: AIDS/HIV problem; Address; Affect; Africa South of the Sahara; Antibodies; Avidin; Bedside Testings; Biological Assay; Biotin; Blood; CD4 Positive T Lymphocytes; Cells; Chemistry; Cytolysis; Detection; Development; Devices; Diagnostic; Diagnostic tests; Electric Conductivity; Electrodes; Ensure; Generations; Goals; HIV; Health Personnel; Home visitation; House Call; Human; Individual; Label; Laboratories; Life; Link; Liposomes; Longevity; Lymphocyte Count; Methods; Microfluidic Microchips; Microfluidics; Modeling; Monitor; Patients; Principal Investigator; Property; Quality of life; Resources; Scheme; Solutions; Streptavidin; Surface; Symptoms; Techniques; Testing; Time; Vesicular stomatitis Indiana virus; Viral; Viral Antibodies; Viral Load result; Virion; Virus; Whole Blood; Work; antiretroviral therapy; base; biochip; cost; electric impedance; innovation; interdisciplinary collaboration; novel; particle; point of care; portability; programs; public health relevance; success; treatment strategy; viral detection

DESCRIPTION (provided by applicant): HIV/AIDS affects more than 33 million people throughout the world, and is especially a critical problem in resource-poor regions in sub-Saharan Africa, where 67% of all HIV/AIDS patients live. Antiretroviral therapy (ART) increases the longevity and quality of life for HIV patients, and global efforts have increased the accessibility of such treatment by 30-fold in sub-Saharan Africa between 2003 and 2008. However, the lack of objective diagnostic tests to determine when to start ART and to monitor its success hinders the effective use of treatment. In addition to the counting of CD4+ T Lymphocytes, it is also highly desirable to perform viral load counts at the point-of-care, as both these parameters are needed for the development of the appropriate treatment strategy. HIV viruses could occur at levels ranging from 10pfu/¿l to thousands of pfu/¿l of whole blood, making it challenging to detect these minute quantities of particles. Current tests include both antibody- based and PCR-based and are not available at point-of-care, especially for resource-limited settings. It should be also pointed out that such devices would of course be extremely valuable also for the developed world, for remote settings, at bedside, or at the doctor's office for a range of applications in detection of viruses. As a solution to these problems presented above, micro-fabricated point-of-care (POC) biochips for HIV/AIDS analysis hold tremendous promise. We propose to develop an integrated device for the electrical detection of HIV viral load at point-of-care. We propose to build on our extensive preliminary work to develop electrically- based sensing methods within micro-fluidic biochips to greatly reduce the operating costs, increase portability, and provide simple-to-use diagnostic kits that can be operated by healthcare workers in remote facilities or during home visits. Our integrated approach is innovative as we will; (i) capture the specific viruses from whole blood, (ii) label these viruses using liposomes tagged with antibodies for the viral particles, (iii) lyse the liposomes by lowerin the electrical conductivity of the medium, and detect the changes in the electrical impedance of the medium in the microfluidic capture chamber. The change in impedance is expected to correlate to the number of virus particles captured. We plan to use VSV-G virus spiked in whole human blood as a model for HIV during this R21 to demonstrate the proof of concept of the novel detection scheme. The interdisciplinary collaboration brings together the expertise of the Co-PIs. We will build on the extensive work in Bashir group (UIUC) on development of microfluidic point-of-care tests that are electrically based, and in Lee Lab (UCI) in microfluidics and generation of liposomes of controlled size and properties. With the integration of these two sets of expertise, we expect to address the grand challenge of developing point-of-care viral load assays.

描述（由申请人提供）：艾滋病毒/艾滋病影响着全世界超过 3300 万人，尤其是撒哈拉以南非洲资源匮乏地区的一个严重问题，那里有 67% 的艾滋病毒/艾滋病患者接受抗逆转录病毒治疗。 ART（抗逆转录病毒治疗）可延长艾滋病毒患者的寿命并提高生活质量，2003 年至 2008 年间，全球努力已将撒哈拉以南非洲地区获得此类治疗的机会增加了 30 倍。然而，缺乏客观的诊断测试来确定何时开始 ART 并监测其成功，阻碍了治疗的有效使用。除了 CD4+ T 淋巴细胞计数外，还非常需要在此时进行病毒载量计数。护理，因为两者 制定适当的治疗策略需要这些参数，HIV 病毒的水平可能在 10pfu/¿ l 至数千 pfu/¿ l 的全血，使得检测这些微量颗粒具有挑战性，目前的检测包括基于抗体和基于 PCR 的检测，并且无法在护理点进行，特别是在资源有限的情况下。指出这种设备对于发达国家来说当然也非常有价值，用于远程设置、床边或医生办公室的病毒检测的一系列应用作为上述问题的解决方案，微制造。护理点 (POC)用于艾滋病毒/艾滋病分析的生物芯片具有巨大的前景。我们建议开发一种用于即时检测艾滋病毒病毒载量的集成设备。我们建议在我们广泛的前期工作的基础上开发微型内的基于电的传感方法。 -流体生物芯片可大大降低运营成本，提高便携性，并提供易于使用的诊断套件，可供医护人员在远程设施或家访期间操作。来自全血的特定病毒， (ii)使用标记有病毒颗粒抗体的脂质体来标记这些病毒，(iii)通过降低介质的电导率来裂解脂质体，并检测微流体捕获室中介质的电阻抗的变化。预计与捕获的病毒颗粒数量相关。我们计划在 R21 期间使用掺入人类全血中的 VSV-G 病毒作为 HIV 模型，以证明新检测的概念。该跨学科合作汇集了联合首席研究员的专业知识，我们将在巴希尔小组（UIUC）和李实验室（UCI）开发基于电的微流体即时测试方面的广泛工作的基础上进行。在微流体学中 通过整合这两套专业知识，我们期望能够解决开发即时病毒载量测定的巨大挑战。