The Transfusion Chip: A Simple, Low Cost Microarray for DNA Based Blood Typing

输血芯片：一种简单、低成本的微阵列，用于基于 DNA 的血型分析

• 批准号: 8199053
• 负责人: MICHAEL E. HOGAN
• 金额: $ 11.58万
• 依托单位: GENOMICS USA, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8199053
• 关键词: Adopted; Agglutination; Agglutination Tests; Antigenic Variation; Biochemical; Blood; Blood Group Antigens; Blood specimen; Blood typing procedure; Bypass; Computer software; DNA; DNA Microarray Chip; DNA purification; Dyes; Ensure; Future; Generations; Genes; Genetic; Genetic Variation; Genetic screening method; HIV; Hepatitis B Virus; Label; Manuals; Measures; Microarray Analysis; Minor; Modification; National Institute of Allergy and Infectious Disease; Nucleic Acids; Performance; Phase; Population; Process; Public Health; Reaction; Reagent; Robotics; Safety; Sampling; Screening procedure; Serologic tests; Serological; Site; Small Business Innovation Research Grant; Solutions; Sorting - Cell Movement; Technology; Testing; Transfusion; Validation; Variant; Vascular blood supply; Viral; Work; base; blood group; cost; design; genetic technology; manufacturing scale-up; pathogen; prototype; scale up; trait; trend

DESCRIPTION (provided by applicant): Low-cost nucleic acid analysis, to detect HIV & HBV, has revolutionized screening of the blood supply. In spite of that new genetic sophistication, traditional blood group typing [ABO, Rh] is still performed, for the most part, via variations upon 80-year-old agglutination testing. Recently, the genetic variation which gives rise to the standard blood groups has been defined, and shown to be relatively simple. In parallel, it has been shown that, to enhance transfusion quality, the historical marker set might be expanded to include a more complete panel of 7 traits: [ABO, Rh, Duffy, Kidd, Kell, Dombrock & MNS] for which the underlying genetics are now known. These two parallel trends suggest that genetic testing has emerged as the future of blood group screening: but only if the genetic test can be delivered at roughly the same cost and level of technical simplicity as agglutination, or the current panel of nucleic acid based pathogen tests. We have observed that, with minor modification, the suite of biochemical, hardware & software technologies that we have developed for microarray based HLA-typing on raw blood could, instead, be transformed into low-cost technologies for blood-group typing, which we refer to as "The Transfusion-Chip". In this 6-month Phase I plan, we propose 2 Specific Aims as Milestones to justify a subsequent Phase II. SA1. Design & preliminary validation of 13-15 PCR reactions to be performed in parallel. All informative sites among the set of 7 blood marker genes will be amplified in parallel, to generate a dye-labeled amplicon set that is ready to be used for microarray hybridization. SA2. Microarray probe design, fabrication &preliminary validation. A Transfusion-Chip prototype will be designed and fabricated, to interrogate the informative sequence variations which define blood group variation among the set of 7 loci. The validation will employ amplified DNAs from SA1. Relation to a Follow-on Phase II. Completion of Phase I will yield a prototype PCR-based sample labeling reaction (that works on raw blood) and a prototype low-cost Transfusion-Chip microarray for DNA based blood-typing. Phase II will focus on refinement of the PCR and microarray designs, manufacturing scale-up, preliminary PCR kit fabrication and delivery of those materials to beta testers, comprising 3-4 top blood labs: to compare Transfusion-Chip performance to serological typing and to the competing microarray and PCR tests. Public Health Narrative. Genetic testing for pathogen contamination has revolutionized the safety of the blood supply, yet analogous DNA based testing of the underlying blood groups has lagged-behind, due to cost and complexity. A microarray-based approach to blood group typing that we propose here, will allow blood type analysis to be modernized, by exploiting two decades worth of blood group genetics: allowing analysis of blood group antigen variation, in a way that is more complete than can be obtained by serology, at a cost and level of technical simplicity that is as practical as that of blood pathogen screening. The resulting technology will not only reduce the cost of blood supply testing in the US, but we propose, will enable a new generation of enhanced, DNA-based blood group typing in the developing world. PUBLIC HEALTH RELEVANCE: A microarray-based approach to blood group typing that we propose here, will allow blood type analysis to be modernized, by exploiting two decades worth of blood group genetics: allowing analysis of blood group antigen variation, in a way that is more complete than can be obtained by serology, at a cost and level of technical simplicity that is as practical as that of blood pathogen screening. The resulting technology will not only reduce the cost of blood supply testing in the US, but we propose, will enable a new generation of enhanced, DNA-based blood group typing in the developing world.

描述（由申请人提供）：用于检测HIV和HBV的低成本核酸分析彻底改变了血液供应的筛查。尽管有这种新的基因复杂性，传统的血型分型[ABO，Rh]仍然在很大程度上通过80年前的凝集试验的变化来进行。最近，产生标准血型的遗传变异已经被定义，并且显示出相对简单。与此同时，已经表明，为了提高输血质量，历史标记集可以扩展到包括7个性状的更完整的组：[ABO，Rh，Duffy，Kidd，Kell，Dombrock和MNS]，其基础遗传学现在是已知的。这两个平行的趋势表明，基因检测已经成为血型筛查的未来：但前提是基因检测的成本和技术简单程度与凝集法或目前基于核酸的病原体检测大致相同。我们已经观察到，通过微小的修改，我们为基于微阵列的原始血液HLA分型开发的一套生物化学，硬件和软件技术可以转化为低成本的血型分型技术，我们称之为“输血芯片”。在这个为期6个月的第一阶段计划中，我们提出了2个具体目标作为里程碑，以证明后续第二阶段的合理性。SA 1.并行进行13-15个PCR反应的设计和初步验证。将平行扩增7个血液标志物基因组中的所有信息位点，以产生准备用于微阵列杂交的染料标记的扩增子组。SA 2.微阵列探针设计、制造及初步验证。将设计和制造输血芯片原型，以询问定义7个基因座集合中血型变异的信息序列变异。验证将使用来自SA 1的扩增DNA。 与后续阶段II的关系。第一阶段的完成将产生一个原型PCR为基础的样品标记反应（对原料血）和原型低成本输血芯片的DNA为基础的血型。第二阶段将专注于PCR和微阵列设计的改进，生产规模的扩大，初步PCR试剂盒的制造和交付这些材料的β测试，包括3-4个顶级血液实验室：比较输血芯片的性能与血清学分型和竞争的微阵列和PCR测试。公共卫生叙事。病原体污染的基因检测已经彻底改变了血液供应的安全性，但是由于成本和复杂性，基于DNA的潜在血型的类似检测已经落后。我们在这里提出的基于微阵列的血型分型方法，将通过利用二十年的血型遗传学来实现血型分析的现代化：允许以比血清学更完整的方式分析血型抗原变异，其成本和技术简单性与血液病原体筛查一样实用。由此产生的技术不仅将降低美国血液供应测试的成本，而且我们建议，将使发展中国家的新一代增强型，基于DNA的血型分型成为可能。 公共卫生相关性：我们在这里提出的基于微阵列的血型分型方法，将通过利用二十年的血型遗传学来实现血型分析的现代化：允许以比血清学更完整的方式分析血型抗原变异，其成本和技术简单性与血液病原体筛查一样实用。由此产生的技术不仅将降低美国血液供应测试的成本，而且我们建议，将使发展中国家的新一代增强型，基于DNA的血型分型成为可能。