Digital SlipChip Technology for POC and Resource-Limited Viral Load Measurements

描述（由申请人提供）：艾滋病毒是一个重大的全球健康问题——最近的估计表明，约有3300万人感染了艾滋病毒，其中96%居住在发展中国家。虽然艾滋病毒抗逆转录病毒疗法是有效的，而且越来越容易获得，但有必要对治疗进行定量监测，以便向患者提供适当护理，并控制耐药艾滋病毒菌株在全世界的出现和传播。在资源有限的环境中，迫切需要这样的检测，以避免与耐药性相关的卫生保健费用的惊人增加和转向二线和三线疗法。然而，目前的廉价、定性、是/否诊断工具不能指导治疗或监测耐药性，而且目前基于pcr的定量检测需要昂贵的设备和复杂的环境，不适合资源有限的医疗点（POC）。此外，对大多数POC方法来说，分离和浓缩病毒RNA是具有挑战性的。SlipChip是PI实验室的一项高度创新的技术，它独特地将高水平的性能与简单的制造和操作相结合。该芯片由两个相对移动或“滑动”的板组成，由一种与样品流体不混溶的流体润滑，并提供表面化学控制和防止交叉污染。滑动使两个板中的孔接触或不接触以执行诊断分析，并且可以在同一芯片上执行超过7个数量级的操作（例如从100 pL到1 mL）。SlipChip将有助于整合i)上游样品制备，分离和浓缩病毒RNA； ii)通过使用等温扩增化学试剂的“数字”（单分子）检测，通过核酸扩增对病毒颗粒进行定量；iii)下游信号扩增，实现简单读出。该团队包括PI， co-PI以及各种工业和全球卫生合作者，他们的专业知识涵盖创新微技术的开发，RNA的分子生物学和等温核酸扩增，诊断分析的临床应用，FDA批准的诊断测试的开发和传播，一次性塑料的制造，灌装和包装，以及诊断设备的制造。与美国、印度和尼日利亚的艾滋病患者一起工作。该小组将确保所有研究都是在正确的全球卫生背景下进行的，并且是在可以廉价制造并在资源有限的情况下使用的化学品、一次性用品和设备的背景下进行的。开发用于监测HIV病毒载量的POC设备符合Francis Collins主任所阐述的NIH五个优先事项中的两个：它将对全球健康产生重大影响，并有可能最终降低美国的医疗保健成本。从长远来看，这项工作将使SlipChip平台成为一项“颠覆性创新”，通过降低成本、简化和提高基于核酸的分子诊断的可用性和适用性来改善医疗保健。

DESCRIPTION (provided by applicant): HIV is a significant global health problem- recent estimates suggest ~33 million people living with HIV, with 96% residing in the developing world. While HIV antiretroviral therapy is effective and increasingly available, quantitative monitoring of treatment is necessary to provide proper care to patients, and to control the emergence and spread of drug-resistant strains of HIV worldwide. Such tests are urgently needed for resource-limited settings to avoid staggering increases in health care costs associated with drug resistance and switching to second- and third-line therapies. However, current inexpensive, qualitative, yes/no diagnostic tools cannot guide treatment or monitor drug resistance, and current quantitative PCR-based assays require expensive equipment and complex environments unsuitable for point-of-care (POC) in resource-limited settings. Additionally, isolating and concentrating viral RNA is challenging for most POC approaches. The SlipChip is a highly innovative technology from the PI's laboratory that uniquely combines a high level of performance with simplicity of fabrication and operation. The chip consists of two plates that move- or "slip"- relative to one another, lubricated by a fluid that is immiscible with the sample fluid and also provides control of surface chemistry and prevents cross-contamination. Slipping brings wells in the two plates in and out of contact to execute a diagnostic assay, and manipulations of volume ranging over 7 orders of magnitude (e.g. from 100 pL to 1 mL) can be performed on the same chip. The SlipChip will facilitate integration of i) upstream sample preparation to isolate and concentrate viral RNA with ii) quantification of viral particles via nucleic acid amplification using "digital" (single molecule) detection using isothermal amplification chemistries, with iii) downstream signal amplification to enable simple readout. The team that includes PI, co-PIs and a diverse set of industrial and global health collaborators has expertise spanning development of innovative micro-technologies, to molecular biology of RNA and isothermal nucleic acid amplification, to clinical use of diagnostic assays, to development and dissemination of FDA- approved diagnostics tests, to manufacturing, filling and packaging of plastic disposables, to manufacturing of diagnostic equipment, to working with AIDS patients both in the US and in India and Nigeria. This team would ensure that all research is done in the correct context of global health and in the context of chemistries, disposables, and equipment that can be manufactured inexpensively and used in resource-limited settings. The development of a POC device to monitor HIV viral loading fits within two of the five priorities of the NIH as articulated by Director Francis Collins: it will have a large impact on global health, and it has the potential to ultimately reduce healthcare costs in the US. Long term, this work would lead to SlipChip platform as a "disruptive innovation" to improve healthcare by reducing costs, and simplifying and increasing availability and applicability of nucleic acid-based molecular diagnostics. PUBLIC HEALTH RELEVANCE: Over 33 million people worldwide are living with HIV, the majority of them in resource-limited settings, with anti- retroviral drugs becoming more and more widely available; however, without technologies for monitoring viral load in patients on anti-retroviral drugs, drug resistant strains of HIV will emerge and spread. This work will develop a SlipChip-based platform to measure the HIV viral load at the point of care inexpensively and simply to prevent a global health care catastrophe caused by emergence and spread of drug-resistant HIV. This work is also likely to act in the US as a "disruptive technology" to improve healthcare and reduce costs.