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3300万艾滋病毒携带者，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，而且越来越多地可用，但有必要对治疗进行量化监测，以向患者提供适当的护理，并控制艾滋病毒耐药株的出现和在世界各地的传播。对于资源有限的环境，迫切需要这样的检测，以避免与抗药性和转向二线和三线疗法相关的医疗费用惊人增加。然而，目前廉价的、定性的、是/否的诊断工具无法指导治疗或监测耐药性，而当前基于定量PCR的分析需要昂贵的设备和复杂的环境，不适合在资源有限的情况下进行护理点(POC)。此外，分离和浓缩病毒RNA对于大多数POC方法来说都是具有挑战性的。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结合在一起。该芯片由两块板组成，这两块板相对移动--或“滑动”--由一种与样品液不相容的流体润滑，还提供了表面化学控制和防止交叉污染。滑动使两个板中的井接触和脱离接触以执行诊断分析，并且可以在同一芯片上执行范围超过7个数量级(例如，从100pl到1ml)的体积处理。该芯片将促进将i)上游样品制备以分离和浓缩病毒RNA与ii)通过使用等温放大化学的“数字”(单分子)检测对病毒颗粒进行量化，以及iii)下游信号放大以实现简单读出的集成。该团队包括PI、共同PIs和一系列不同的工业和全球健康合作者，他们拥有专业知识，包括创新微技术的开发，RNA分子生物学和等温核酸放大，诊断分析的临床应用，FDA批准的诊断测试的开发和传播，塑料一次性用品的制造、灌装和包装，诊断设备的制造，以及与美国、印度和尼日利亚的艾滋病患者合作。该小组将确保所有研究都在全球健康的正确背景下进行，并在化学物质、一次性用品和设备的背景下进行，这些化学物质、一次性物品和设备可以廉价制造并在资源有限的环境中使用。开发一种用于监测艾滋病毒病毒载量的POC设备符合美国国立卫生研究院主任弗朗西斯·柯林斯所阐述的五项优先事项中的两项：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.