A Centrifugal Microfluidics Based Pathogen Analyzer

描述(由申请人提供)：生物战剂(BWA)攻击的潜在威胁以及SARS(严重急性呼吸系统综合症)病毒向二十多个国家的传播，以及最近禽流感病毒在全球传播的威胁，强调迫切需要开发诊断系统，能够迅速识别具有潜在流行病学后果的危险传染病的出现。目前基于DNA的便携式感染性病原体检测系统缺乏多样性，即同时分析大量病原体或它们的基因，它们不能很好地集成或自动化样品制备方法，不能在现场或护理点应用中有效使用。微阵列平台实际上不会限制对大量病原体的多路检测以及多个基因对它们的表征。第一阶段项目成功地展示了使用离心式微流控技术制备样品的几种创新方法。设计和制造了三维或多层离心盘(3DCD)，为不同的样品制备技术提供了几乎无限的表面积。在第一阶段项目中演示了几种用于3DCD格式的样品制备的创新工具。第二阶段项目的总体目标是整合这些工具，并开发用于检测传染病和/或生物战剂的特定化验的CD。DNA微阵列检测平台将与3D CD上的样品制备集成在一起。早些时候开发并商业化的电子微阵列技术将用于检测。这项技术利用凝胶驱动的DNA传输和微阵列上的杂交确保了快速的DNA检测时间。第二阶段计划的具体目标是开发带有嵌入式工具的3DCD技术，以完成对几种典型的呼吸道传染病病原体以及生物战剂的样品制备和检测。一项完全开发的技术将产生一个小型便携式或护理点系统，带有可更换的CD，允许选择病原体检测面板，包括呼吸道传染病、生物战剂或紧急病毒面板。

DESCRIPTION (provided by applicant): Potential threat of biological warfare agent (BWA) attacks as well as spread of SARS (Severe Acute Respiratory Syndrome) virus to more than two dozen countries and a recent threat of global spread of bird flu virus emphasized the need for urgent development of diagnostic systems which will be capable to rapidly recognize emergence of dangerous infectious diseases with potential epidemiologic consequences. Current DNA based portable systems for the detection of infectious agents lack multiplexing, i.e., simultaneous analysis of large number of pathogens or their genes, they do not integrate well or automate the sample preparation methods and cannot be used efficiently in the field or in point-of care applications. The microarray platform practically does not pose a limit to multiplexed detection of large number of pathogens as well as their characterization by multiple genes. The Phase I project successfully demonstrated several innovative approaches to sample preparation using centrifugal microfluidics. Three dimensional or multilayer centrifugal discs (3DCD) have been designed and fabricated which offer practically unlimited surface area for implementation of different sample preparation techniques. Several innovative tools for sample preparation in the 3DCD format have been demonstrated in the Phase I project. The overall goal of the Phase II project is to integrate these tools and develop CDs for particular assays for the detection of infectious disease and/or biological warfare agents. A DNA microarray detection platform will be integrated with the sample preparation on the 3DCD. An electronic microarray technology developed and commercialized earlier, will be used in detection. This technology assures fast DNA detection times using electrophoretically driven DNA transport and hybridization on the microarray. Specific aims of the Phase II program is to develop 3DCD technology with embedded tools for complete sample preparation and detection of several representative respiratory infectious disease pathogens as well as biological warfare agents. A fully developed technology will yield a small portable or point-of-care system with exchangeable CDs allowing choice of the pathogen detection panels including respiratory infectious disease, biological warfare agents or emergent viruses panels.