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

描述(申请人提供)：艾滋病毒大流行对世界上最贫穷地区的影响不成比例，因此，生活在3400万人中的很大一部分人 世界各地的艾滋病患者无法获得适当的医疗服务。在这些人群中进行适当治疗的一个主要障碍是无法获得监测疾病进展和评估抗逆转录病毒治疗效果的诊断工具。由于成本、大小和对熟练操作员的需要，标准的仪器设备通常是不实用的，因此迫切需要低成本、用户友好的护理点诊断测试。特别是，在早期发现感染、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方面，技术是非常必要的。 该项目旨在设计两个芯片实验室生物传感器平台，能够定量检测血浆中的艾滋病毒和感染的CD4+T细胞中的前病毒。这些平台将成为低成本、用户友好的护理点检测设备的基础，这些设备可以分析少量的全血输入，以确定艾滋病毒携带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的病毒特异性抗体从全血样本中捕获血浆中的整个病毒颗粒来检测它们。在冲洗掉血细胞和其他碎片后，捕获的病毒将被含有浓缩磷酸盐缓冲盐水(PBS)的脂质体标记。背景介质会被低电导率介质冲洗掉，通过提高装置温度，刺激离子的释放，引发脂质体颗粒的泄漏，这些离子的释放可以通过阻抗谱进行定量。病毒数量将从阻抗的变化中确定。 同样，我们检测前病毒的方法是从全血样本中捕获微流体室中的CD4+T淋巴细胞。随后用化学试剂裂解，将细胞内的内容物洗入反应室，在那里可以进行分析。通过聚合酶链式反应扩增病毒DNA(前病毒)，或最终用电子传感器检测，不需要光学元件，是低成本和快速检测的极佳平台。 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，使治疗更知情，在偏远和资源有限的环境下更容易获得频繁的诊断数据。

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.