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（由适用提供）：艾滋病毒/艾滋病影响着全球超过3,300万人，在撒哈拉以南非洲贫困地区尤其是一个关键问题，所有艾滋病毒/艾滋病患者中有67％居住。抗逆转录病毒疗法（ART）增加了HIV患者的寿命和生活质量，全球努力在2003年至2008年间，在撒哈拉以南非洲地区的30倍增加了这种治疗的可访问性。但是，缺乏客观诊断测试来确定何时开始ART并确定其成功促进其成功的治疗方法。除了对CD4+ T淋巴细胞的计数外，也非常需要在护理点上进行病毒载荷计数 制定适当的治疗策略需要这些参数。 HIV病毒可能在10pfu/|至数千个PFU/»全血的水平上发生，因此检测到这些分钟数量的颗粒的挑战。当前的测试包括基于抗体和基于PCR的测试，并且在护理点上不可用，尤其是对于资源有限的设置。还应指出，对于发达国家，远程设置，在床边或医生办公室，对于一系列检测病毒的应用，这些设备当然也非常有价值。为了解决上述这些问题的解决方案，用于HIV/AIDS分析的微实验点（POC）生物芯片具有巨大的希望。我们建议开发一个集成装置，用于在护理点上对HIV病毒负荷进行电气检测。我们建议以我们广泛的初步工作为基础，以开发微流体生物芯片中的电气感应方法，以大大降低运营成本，提高可移植性并提供易于使用的诊断套件，这些诊断套件可以由远程设施或家庭访问期间的医护人员可以操作。我们的综合方法是创新的。 （i）从全血中捕获特定病毒，（ii）使用用抗体标记的病毒颗粒标记的脂质体标记这些病毒，（iii）通过降低培养基的电导率来裂解脂质体，并检测微流体捕获室中培养基电气障碍的变化。与捕获的病毒颗粒数量相关。我们计划在此R21期间使用在整个人类血液中尖刺的VSV-G病毒作为HIV的模型，以证明新型检测方案的概念证明。跨学科的合作汇集了Co-Pis的专业知识。我们将基于Bashir组（UIUC）的广泛工作，以开发电气点测试，这些测试是基于电的，以及在Microfluidics中的Lee Lab（UCI）中 以及受控大小和特性的脂质体的产生。随着这两套专业知识的整合，我们希望应对开发护理点病毒负荷分析的巨大挑战。