Nanomagnetic isolation and sensing for mobile HIV-1 self-testing

项目摘要 抗逆转录病毒治疗（ART）需要密切监测HIV-1血浆病毒血症， 抑制，并确定由于不依从或耐药性引起的病毒反弹，以启动 及时调整治疗方案。因此，经常有效地监测血液中的艾滋病毒， 可用于家庭自我检测的设备是一个关键需求。仅限当前的自我检测策略 检测抗体，现有的病毒检测方法需要大量的设备， 专业知识（RNA）和/或具有有限的敏感性（抗原）。我们的目标是填补这一空白， 开发一种廉价、自动化、便携式系统， 血浆病毒血症具有高特异性和足够的灵敏度，可用于这些患者的自我监测。 contexts.本项目的最终目标是开发一种低成本（< 2美元/检测）用于人类粗样品（血浆）的即时核酸诊断， 定量病毒RNA，检测限< 10个病毒/样品（相当于临床相关的 在10 μ l手指刺血上1000拷贝/ml的阈值）。该方法将易于执行， 不需要复杂的扩增程序如PCR。我们方法的新奇在于 将我们团队发明的两项创新技术联合收割机结合起来-一项用于目标分离， 一种浓度适用于任何起始血液体积，一种浓度适用于无标记核酸 无需放大的检测-集成到集成设备中。我们的R66的目标是设计一个 用于演示组织上HIV病毒载量定量的基于实验室的集成系统 将培养物衍生的病毒掺入健康血浆中。我们R33的目标很先进 我们的技术发展成为一个完全自动化的手持移动电话为基础的设备， 由非微流体专家使用，并通过使用库存临床样本进行评估， 与宾夕法尼亚大学艾滋病研究中心合作。

Project Summary Antiretroviral treatment (ART) requires close monitoring of HIV-1 plasma viremia to confirm viral suppression, and to identify viral rebound due to nonadherence or drug resistance to initiate timely treatment modification. Thus, frequent efficient monitoring for HIV in blood via methods that are accessible for home self-testing is a critical need. Current self-detection strategies only detect antibody, and existing virus detection methods require extensive equipment and expertise (RNA) and/or have limited sensitivity (antigen). We aim to fill this gap with the development of an inexpensive, automated, portable system that will detect new or recurrent plasma viremia with high specificity and sufficient sensitivity for self-monitoring in these contexts. The ultimate goal of this project is to develop a platform technology for low-cost (< $2/test) point-of-care nucleic acid diagnostics for use on crude human samples (plasma) to quantitate viral RNA with a detection limit < 10 virus / sample (equivalent to a clinically-relevant threshold of 1000 copy/ml on 10 ul finger prick blood). The method will be simple to perform and will not require complex amplification procedures such as PCR. The novelty of our approach is to combine two innovative technologies invented by our team - one for target separation and concentration suitable for any starting blood volume, and one for label-free nucleic acid detection without amplification – into an integrated device. The goal of our R66 is to design a laboratory-based integrated system to demonstrate quantitation of HIV viral load on tissue culture derived virus spiked into healthy plasma. The goals of our R33 are advanced development of our technology into a fully-automated handheld mobile-phone based device to be used by non-microfluidic experts, and evaluation using banked clinical samples through partnership with Penn's Center for AIDS Research.