Nanodroplet PCR Lab-Chip for Clinical Pathogen Detection

描述（由申请人提供）：据估计，每年住院患者获得200万例医疗保健相关感染，导致9万例死亡和约45亿美元的额外医疗保健费用。在这些感染中，很大一部分是由金黄色葡萄球菌引起的，特别是耐甲氧西林的金黄色葡萄球菌。金黄色葡萄球菌（MRSA）。已经开发了基于PCR的快速检测用于在卫生保健环境中筛查MRSA，但是现有核酸检测平台的高成本和复杂性限制了它们在大多数卫生保健环境中用于常规筛查的接受度。该项目的目标是开发一种快速，低成本，易于使用的核酸检测平台，可用于广泛的医疗保健环境中的MRSA筛查。拟议的测试平台基于Advanced Liquid Logic的“数字微流体”技术，该技术使用形成在低成本和一次性印刷电路板基板上的电极阵列实现亚微升大小的液滴的可编程微操作。在该项目的第一阶段，Advanced Liquid Logic开发了基于芯片的方法，用于快速热循环、实时PCR检测、基于磁珠的样品制备以及反应制备和混合。在第二阶段，这些结果将作为开发用于近患者和床旁使用的产品原型核酸分析系统的基础。MRSA筛查测试将在该平台上开发，并在准备FDA提交时进行广泛验证。最终，可以开发更多的检测盒，用于筛查和检测其他医疗保健相关感染。这种芯片实验室平台将在灵活性、可扩展性、性能和成本方面提供优于其他方法的显著优势。 公共卫生相关性：将开发一种检测耐甲氧西林金黄色葡萄球菌的系统，以帮助控制耐甲氧西林细菌在医院环境中的传播。

DESCRIPTION (provided by applicant): It is estimated that 2 million health-care associated infections are acquired each year by hospitalized patients resulting in 90,000 deaths and about $4.5 billion in excess health care costs. A large and growing percentage of these infections can be attributed to Staphylococcus aureus and, in particular, to methicillin-resistant S. aureus (MRSA). Rapid PCR-based tests have been developed for screening MRSA in health-care settings, but the high cost and complexity of existing nucleic acid testing platforms have limited their acceptance for routine screening in most health-care settings. The goal of this project is to develop a rapid, low-cost, easy-to-use nucleic acid testing platform that can be deployed for MRSA screening in a wide range of health-care settings. The proposed testing platform is based on Advanced Liquid Logic's "digital microfluidics" technology which enables programmable micro-manipulation of sub-microliter sized droplets using an electrode array formed on a low-cost and disposable printed circuit board substrate. In Phase I of this project Advanced Liquid Logic developed chip-based methods for rapid thermocycling, real-time PCR detection, magnetic-bead based sample preparation, and reaction preparation and mixing. In Phase II, these results will serve as the foundation for the development of a product prototype nucleic acid analysis system for near-patient and point-of-care use. An MRSA screening test will be developed on the platform and extensively validated in preparation for an FDA submission. Ultimately, additional cartridges could be developed for screening and detection of other health-care associated infections. This lab-on-a-chip platform will offer significant advantages over other approaches in terms of flexibility, scalability, performance and cost. PUBLIC HEALTH RELEVANCE: A system for the detection of methicillin-resistant Staphylococcus aureus will be developed to help control the spread of antibiotic-resistant bacteria in hospital settings.