The Transfusion Chip: Phase II Technology Validation

描述(申请人提供)：血型鉴定是整个个性化医疗领域的鼻祖。在最近授予的为期6个月的第一阶段中，我们提议迈出开发一种极低成本的微阵列测试的第一步--“输血芯片”(T-Chip)，它将历史上通过血清学进行血型鉴定的做法转变为一种简单、廉价的DNA测试。在这里，在这个后续的第二阶段，我们展示了在实现关键的第一阶段里程碑方面取得的巨大成功：我们已经开发和优化了22种聚合酶链式反应，包括整个7基因血型基因集[ABO、Rh、Duffy、Kidd、Kell、Dombrock和Mns]；我们已经制造了能够分析7基因集中所有已知遗传变异的输血芯片芯片原型；并且通过在提交第一阶段报告时我们还没有发明的技术，我们已经开发出了原始Genusa微阵列测试的新变体，将把T芯片的制造成本降低到每个微阵列1美元的价值。在第一阶段，我们还展示了未经纯化的Oragene TM稳定的原始唾液可以用作T-Chip上基于DNA的血型分析的基础。我们认为，Oragene TM稳定的原始唾液采集与低成本T-Chip相结合可以成为一种独特的技术组合：因为它将允许在家中自行采集DNA样本，然后将低成本的环境温度运往中央实验室，然后在T-芯片上进行非常低成本、非常高吞吐量的血型鉴定。这一人口规模血型测定模式将是本阶段第二阶段的发展重点：因此，我们建议实现人口规模血库和输血医学的全新愿景，并在此过程中，将使血型测定在21世纪重新成为遗传个性化医学领域的模式和黄金标准：就像它在100年前首次做到的那样。基于人口规模血型基因检测的模型，我们提出了以下第二阶段的目标。SA1.完成并验证输血芯片的多重PCR前端，用于DNA样本。SA2.完成并验证输血芯片的多重聚合酶链式反应前端，用于原材料。SA3.优化和验证输血芯片的制造，每张幻灯片48个阵列“$1微阵列”格式。SA4.修改现有Genusa分析软件RicimerTM，以实现自动血型鉴定。SA5.验证用于纯化的DNA、原始血液和Oragene TM稳定的原始唾液样本的输血芯片。第二阶段交付成果和商业化计划。在第二阶段完成后，我们将为后续的Beta测试准备好输血芯片试剂盒原型(微阵列、聚合酶链式反应试剂、杂交试剂、专用软件)。这种试剂盒将在制造控制(国际标准化组织13845)下准备，类似于Genusa的人类白细胞抗原芯片产品，该产品目前正投放到组织兼容性和免疫遗传市场。在第二阶段完成后，Genusa将向Ruo血型市场推出T-Chip，同时准备FDA 510(K)申请进入诊断市场，要么单独进入诊断市场，要么与那些在输血药物或家庭样本采集方面有公认市场存在的公司合作，或者两者兼而有之。

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.