Nanodroplet PCR Lab-Chip for Clinical Pathogen Detection

描述(申请人提供)：据估计，每年有200万名住院患者感染与医疗保健有关的感染，导致90,000人死亡，额外医疗费用约为45亿美元。这些感染中有很大一部分是由金黄色葡萄球菌引起的，尤其是耐甲氧西林金黄色葡萄球菌(MRSA)。基于聚合酶链式反应的快速检测已被开发用于在医疗保健环境中筛查MRSA，但现有核酸测试平台的高成本和复杂性限制了它们在大多数医疗保健环境中进行常规筛查的接受度。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，可以在广泛的医疗保健环境中部署用于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产品原型核酸分析系统的基础，供近距离患者和护理地点使用。将在该平台上开发MRSA筛查测试，并在为FDA提交的准备工作中进行广泛验证。最终，可以开发更多的试剂盒，用于筛查和检测其他与卫生保健相关的感染。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比其他方法更大的优势。 公共卫生相关性：将开发一个检测耐甲氧西林金黄色葡萄球菌的系统，以帮助控制耐药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.