The Transfusion Chip: Phase II Technology Validation

描述（由申请人提供）：血型是整个个性化医疗领域的先驱。在最近授予的为期6个月的第一阶段，我们建议在开发一种极低成本的微阵列测试中迈出第一步，“输血芯片”（T-Chip）将历史上通过血清学进行血型定型的做法转变为简单廉价的DNA测试。在这里，在接下来的第二阶段，我们在实现关键的第一阶段里程碑方面取得了实质性的成功：我们已经开发和优化了22个PCR反应，包括整个7个基因血型基因集[ABO，Rh，Duffy，Kidd，Kell，Dombrock和MNS];我们已经制造了一个输血芯片微阵列原型，能够分析7个基因集中所有已知的遗传变异;通过我们在提交第一阶段申请时尚未发明的技术，我们已经开发了原始GenUSA微阵列测试的新变体，该变体将使T芯片的制造成本降低到每个微阵列1美元。 在第一阶段中，我们还证明了未纯化的OrageneTM稳定唾液可以用作T芯片上基于DNA的血型鉴定的基础。我们认为，将OrageneTM稳定的原始唾液收集与低成本的T芯片结合起来可能成为一种独特的技术配对：因为它允许在家中自行收集DNA样本，然后以低成本的环境温度运送到中心实验室，然后在T芯片上进行非常低成本，非常高通量的血型定型。这种人口规模的血型定型模型将是第二阶段的发展重点：因此，我们提出，人口规模血库和输血医学的全新愿景，并在此过程中，将使血型定型在21世纪世纪重新成为基因个性化医学领域的模型和黄金标准：就像它在一百年前首次做到的那样。基于人群规模血型基因检测的模型，我们提出了以下II期目标。SA 1.完成并将多重PCR前端连接到输血芯片，用于DNA样本。SA 2.完成并将多重PCR前端连接到输血芯片，用于原始样本。SA 3.优化和验证输血芯片的制作，每个载玻片48个阵列，“1美元微阵列”格式。SA 4.修改现有的GenUSA分析软件Ricimer TM，用于自动血型分型。SA 5.为纯化的DNA、原始血液和原始OrageneTM稳定的唾液样本提供输血芯片。 第二阶段可销售产品和商业化计划。在第二阶段完成后，我们将准备一个输血芯片套件原型（微阵列，PCR试剂，杂交试剂，专业软件）用于随后的Beta测试。该试剂盒将在生产控制（ISO 13845）下制备，类似于GenUSA的HLA芯片产品，该产品目前正在进入组织相容性和免疫遗传学市场。在第二阶段完成后，GenUSA将向RUO血型检测市场推出T-Chip，并同时准备FDA 510（k）申报，以单独或与在输血医学或家庭样本采集方面具有成熟市场地位的公司合作进入诊断市场。

DESCRIPTION (provided by applicant): Blood-Typing is the progenitor of the entire field of personalized medicine. In the recently awarded 6-month Phase I, we proposed to take the first step in the development of an extremely-low-cost microarray test, "The Transfusion-Chip" (T-Chip) which convert the historical practice of Blood-Typing by serology into a simple inexpensive DNA test. Here, in this follow-on Phase II, we demonstrate substantial success in achieving key Phase I milestones: we have developed and optimized 22 PCR reactions, comprising the entire 7 gene blood group gene set [ABO, Rh, Duffy, Kidd, Kell, Dombrock & MNS]; we have fabricated a Transfusion-Chip microarray prototype capable of analyzing all known genetic variation in the 7 gene set; and via technology that we had not yet invented when we filed the Phase I, we have developed a new variant of the original GenUSA microarray test which will drop the manufacturing cost of T-Chips to values as low as $1 per microarray. In Phase I we have also shown that the raw, unpurified OrageneTM-stabilized saliva can used as the basis for DNA based Blood-Typing on the T-Chip. We feel that the coupling of raw OrageneTM-stabilized saliva collection with the low cost T-Chip could become a unique technology pairing: because it would allow self-collection of DNA samples, at home, then low- cost ambient temperature shipping to centralized labs, to be followed by very-low-cost, very high throughput Blood-Typing on T-Chips. This model for population scale Blood-Typing will be the development focus in this Phase II: thereby enabling, we propose, an entirely new vision for population-scale blood banking & transfusion medicine, and in the process, will allow Blood- Typing to return, in the 21st century, to prominence as the model and gold standard for the field of genetically personalized medicine: as it first did a hundred years ago. Based on that model for population-scale blood group genetic testing, we propose the following set of Phase II Aims. SA1. Complete & Validate the Multiplex PCR Front-end to the Transfusion-Chip, for DNA Samples. SA2. Complete & Validate the Multiplex PCR Front-end to the Transfusion-Chip, for Raw Samples. SA3. Optimize & validate fabrication of the Transfusion-Chip in the 48 array per slide "$1 microarray" format. SA4. Modify existing GenUSA analysis software, RicimerTM, for automated blood group typing. SA5. Validate the Transfusion-Chip for purified DNA, raw blood & raw OrageneTM-stabilized saliva samples. Phase II Deliverables & Commercialization Plan. Upon completion of this Phase II, we will have ready a Transfusion-Chip kit prototype (microarrays, PCR reagents, Hybridization Reagents, Specialized Software) for subsequent Beta Testing. This kit will be prepared under manufacturing control (ISO 13845) similar to the HLA-Chip products from GenUSA which are currently being launched into tissue compatibility and immunogenetics markets. Subsequent to completion of Phase II, GenUSA will launch the T-Chip into the RUO Blood-Typing market and prepare simultaneously for FDA 510(k) submission to enter into the diagnostics market either alone, or in partnership with those with a proven market presence in transfusion medicine or at-home sample collection, or both.