Microfluidic Controlled Gel-Drop Microarrays for Biothreat Diagnostic Devices

用于生物威胁诊断设备的微流控凝胶滴微阵列

• 批准号: 7288668
• 负责人: CHARLES DAITCH
• 金额: $ 56.34万
• 依托单位: AKONNI BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7288668
• 关键词: Agar; Anthrax disease; Bacillus anthracis; Bacillus cereus; Bacteria; Biological Assay; Bioterrorism; Buffers; Cells; Characteristics; Chemistry; Classification; Clinical; Community Hospitals; Compatible; Condition; DNA; Data; Data Analyses; Detection; Development; Devices; Diagnostic; Discrimination; Disease; Drops; Epidemiologic Studies; Fluorescence; Gel; Genes; Genetic; Genomics; Glass; Goals; Hand; Immunologic Adjuvants; Label; Laboratories; Laboratory Study; Letters; Licensing; Life; Liquid substance; Measures; Microarray Analysis; Microfluidics; Nucleic Acid Amplification Tests; Oligonucleotide Probes; Oligonucleotides; Operating System; Optics; Organism; Pattern; Performance; Phase; Phenotype; Phylogenetic Analysis; Polymerase Chain Reaction; Preparation; Procedures; Protocols documentation; Reaction; Reagent; Relative (related person); Reporting; Research; Resources; Ribosomal RNA; Robotics; Sampling; Schedule; Silicon Dioxide; Sister; Slide; Solutions; Specific qualifier value; Surface; Suspension Culture; System; Technology; Temperature; Testing; Tube; Validation; base; biodefense; biosafety level 2 facility; biosafety level 3 facility; biothreat; cost; design; instrument; member; meter; method development; microbial; novel diagnostics; pathogen; pathogenic bacteria; point of care; rRNA Genes; research and development; research study; response; success; therapeutic vaccine

DESCRIPTION (provided by applicant): Whether in clinical or epidemiological settings, there is a continued need in identification of B. anthracis and unambiguous discrimination it from close relatives and organisms with similar characteristics. Genomic analysis provides direct information about tested organism in opposite to indirect, phenotype analysis, however represents in current certified Anthrax tests by complicated (e.g. 16S rRNA gene sequencing), or unreliable (PCR-related) tests and may be performed now at advanced detection laboratories only. Akonni proposes microarray platform that will fill a fundamental technology gap between traditional clinical or disease diagnostics and new biodefense and associated epidemiology research. The objective of this application is to develop a portable clinical oriented diagnostic to assist in the rapid and reliable identification of B. anthracis and relative pathogens from B. cereus group by microarray analysis of pure cultures obtained for sample testing at Laboratory Response Network for Bioterrorism (LRN). Simple, rapid, reliable and low- cost Akonni diagnostic will bring sophisticated and final testing from Level C-D, advanced high-tech laboratories, to Level A, laboratories typical for community hospitals. Akonni will unify benefits of 3D gel drop microarray technology with microfluidic platform, newest 1 min DNA/RNA labeling-fragmentation chemistry and last developments in phylogenetic classification of B. anthracis and its close relatives. The research of Phase 1 will focus on the method development and validation of the gel-drop microarray microfluidic card components. Specific aims include integration of sample preparation protocol in microfluidic card, design, produce and test a gel-drop microarray on a glass slide for identification of B. anthracis and testing of gel- drop microarray against samples prepared on microfluidic card from specified isolates. Final card will accept ~50uL of pure bacterial culture suspension; the card will mix the sample represented by one bacterial colony grown on agar plate with the buffers and provide rRNA isolation, labeling, fragmentation and purification. A pettier heater will control the temperature conditions within the card. The microfluidic card will handle all mixing, metering and washing steps and will be optically compatible for fluorescence detection and data analysis after integration of microarray into the card on Phase 2 of the project. We will validate our diagnostic in our BSL-2 facilities in Argonne National Laboratory.

描述（由申请方提供）：无论是在临床还是流行病学环境中，都需要持续识别B。炭疽菌，并明确区分它与近亲和具有相似特征的生物。基因组分析提供了关于测试生物体的直接信息，而不是间接的表型分析，然而，在目前经过认证的炭疽测试中，基因组分析是通过复杂的（例如16 S rRNA基因测序）或不可靠的（PCR相关的）测试进行的，并且现在只能在先进的检测实验室进行。Akonni提出的微阵列平台将填补传统临床或疾病诊断与新生物防御和相关流行病学研究之间的基本技术差距。本申请的目的是开发一种便携式面向临床的诊断，以帮助快速可靠地识别B。炭疽菌及相关病原菌来自B.蜡样芽胞杆菌群通过微阵列分析的纯培养物获得的样品测试在实验室响应网络生物恐怖主义（LRN）。简单，快速，可靠和低成本的Akonni诊断将带来复杂的和最终的测试，从C-D级，先进的高科技实验室，到A级，社区医院的典型实验室。Akonni将把3D凝胶滴微阵列技术与微流控平台、最新的1分钟DNA/RNA标记-片段化化学和B系统发育分类的最新进展结合起来。炭疽菌及其近亲第一阶段的研究将集中在凝胶滴微阵列微流控卡组件的方法开发和验证。具体目标包括将样品制备方案集成到微流控卡中，设计、制备和测试用于鉴定B的载玻片上的凝胶滴微阵列。炭疽和测试凝胶滴微阵列对样品制备的微流控卡从指定的菌株。最终卡将接受约50 uL的纯细菌培养物悬液;卡将琼脂平板上生长的一个细菌菌落代表的样品与缓冲液混合，并提供rRNA分离、标记、片段化和纯化。一个pettier加热器将控制卡内的温度条件。微流控卡将处理所有混合、计量和洗涤步骤，并在项目第2阶段将微阵列集成到卡中后，光学兼容荧光检测和数据分析。我们将在阿贡国家实验室的BSL-2设施中验证我们的诊断。