Nanodroplet PCR Lab-Chip for Clinical Pathogen Detection

描述（由申请人提供）：据估计，住院病人每年感染200万例与保健有关的感染，造成9万人死亡，保健费用超支约45亿美元。这些感染的很大比例和不断增长的比例可归因于金黄色葡萄球菌，特别是耐甲氧西林金黄色葡萄球菌（MRSA）。基于pcr的快速检测已被开发用于在卫生保健机构中筛查MRSA，但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机构中的常规筛查。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，可以在广泛的医疗保健环境中用于MRSA筛查。提出的测试平台基于Advanced Liquid Logic的“数字微流体”技术，该技术可以使用在低成本和一次性印刷电路板基板上形成的电极阵列对亚微升大小的液滴进行可编程微操作。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护理点使用的产品原型核酸分析系统的基础。MRSA筛选测试将在该平台上开发并广泛验证，以准备向FDA提交。最终，可以开发更多的药筒，用于筛选和检测其他保健相关感染。与其他方法相比，这种芯片实验室平台在灵活性、可扩展性、性能和成本方面具有显著优势。

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.