Biodosimetry with a fully integrated biochip

电离辐射照射使许多基因的表达产生剂量依赖性变化，这可能提供一种评估辐射照射和剂量的手段。我们最近报道了一种完全独立的生物芯片的开发，该芯片由微流体混合器、阀门、泵、通道、腔室、加热器和DNA微阵列传感器组成，用于对复杂的生物样品溶液（如血液）进行DMA分析。我们建议将此装置发展成为一个独立的辐射生物剂量计，适用于大规模的筛选。该盒将包含其自身的电源、控制电子器件、功能性微流体组件和试剂。检测完成后，将检测盒插入读取器，在几秒钟内读取信号。该项目的具体目标是： 1.开发一个从手指刺血收集的模块。 2.用集成寡核苷酸阵列制造和验证环烯烃共聚物生物通道芯片。 3.开发和性能测试集成微/纳米流体盒，用于提取和处理 来自全血的RNA 4.纳入功能基因组学核心鉴定的辐射相关基因表达特征 定制的微流控微阵列。 5.将血样采集装置、RNA提取装置和杂交装置集成在一起， 将微通道器件组装成一个独立的生物芯片，并验证了该生物芯片的性能。 6.集成并评估独立检测盒的试剂储存选项。 7.开发集成控制电子设备，创建适合大规模生产的便携式仪器， 应用于高通量筛选，并验证该设备符合产品要求。 总之，我们将开发能够快速测量一百个或更多基因表达水平的独立生物芯片，这些基因将确定辐射暴露、剂量和损伤。 这种方法只需要从手指上刺一滴血，并且在发生放射性事件时可以随时部署用于大规模人群筛查。

Exposure to ionizing radiation produces dose-dependent changes in the expression of many genes, potentially providing a means to assess both radiation exposure and dose. We have recently reported development of a completely self-contained biochip that consists of microfluidic mixers, valves, pumps, channels, chambers, heaters, and DNA microarray sensors to perform DMA analysis of complex biological sample solutions, such as blood. We propose to develop this device into a self-contained radiation biodosimeter suitable for large scale screening. The cartridge will contain its own power source, control electronics, functional microfluidic components and reagents. After assay completion, the cartridge is inserted into a reader where the signal is read out within seconds. The Specific Aims of this Project are to: 1. Develop a module for the collection of blood from a finger prick. 2. Fabricate and validate cyclo-olefinic co-polymer biochannel chips with integrated oligonucleotide arrays. 3. Develop and performance-test an integrated micro/nano fluidic cartridge for extraction and processing of RNA from whole blood. 4. Incorporate the radiation-related gene-expression signature identified by the Functional Genomics Core into customized microfluidic micro-arrays. 5. Integrate the blood sample collection device, the RNA extraction device and the hybridization microchannel device into a single self-contained biochip, and validate the performance of this biochip. 6. Integrate and evaluate reagent storage options for the self-contained cartridge. 7. Develop integrated control electronics, creating a portable instrument suitable for mass-production and application to high-throughput screening, and verification that this device meets product requirements. In summary, we will develop self-contained biochips capable of rapidly measuring expression levels of a hundred or more genes that will define radiation exposure, dose and injury. This approach will only require a drop of blood from a finger prick, and will be readily deployable for large-scale population screening in the event of a radiological incident.