Biodosimetry with a fully integrated biochip using gene expression signatures

暴露在电离辐射下会使许多基因的表达发生剂量依赖性的变化，这可能为评估辐射暴露和剂量提供了一种手段。我们最近报告了一种完全自给自足的生物芯片的开发，该芯片由微流控混合器、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本溶液(如血液)进行DMA分析。我们建议将该装置发展成适合大规模筛查的自给式辐射生物剂量计。墨盒将包含自己的电源、控制电子设备、功能微流控部件和试剂。分析完成后，将试剂盒插入读取器，在几秒钟内读出信号。该项目的具体目标是： 1.开发用于手指刺伤采血的模块。 2.制备和验证集成寡核苷酸阵列的环烯类共聚生物通道芯片。 3.研制和性能测试了一种用于提取和加工黄曲霉毒素的微/纳米一体化小柱 全血中的核糖核酸。 4.整合功能基因组学核心鉴定的辐射相关基因表达特征 制作成定制的微流控微阵列。 5.集成血样采集器、RNA提取装置和杂交 将微通道器件集成到单个独立的生物芯片中，并验证了该生物芯片的性能。 6.整合并评估自给式药筒的试剂储存选项。 7.开发集成控制电子产品，创造适合批量生产和生产的便携式仪器 应用于高通量筛选，并验证该设备满足产品要求。 总而言之，我们将开发独立的生物芯片，能够快速测量100个或更多基因的表达水平，这些基因将定义辐射暴露、剂量和伤害。 这种方法只需要从手指上扎一滴血，而且在发生辐射事故时，可以随时部署进行大规模人口筛查。

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.