A Parallel High-Throughput Pharmacogenetic Profiling Assay for Clinical Use

描述(由申请人提供)：临床使用的平行高通量药物遗传图谱分析药物基因组学研究已经确定了一些基因，它们的SNP基因型或单倍型与不同的个体药物反应、其他代谢过程或疾病易感性相关。因此，快速准确地确定医学相关区域的基因类型的能力对于了解个人基因特征对这些过程的影响以及对预测性、预防性和个性化医学的发展至关重要。Maxwell Sensors Inc.建议开发条形码蚀刻可评估微珠(BEAM BEAD)技术，该技术将数字条形码微珠和分子化学相结合，用于药物遗传学SNP基因分型分析，可用于高通量的个体分子图谱分析。数字束珠是顺磁性的，通过光刻制作成具有数千个识别码，并可以与核酸、蛋白质或其他小分子功能化，在均相或非均相介质中进行大型多路分析。使用束珠进行基因分型具有灵活性、高通量、易于制造、低成本大规模生产和小体积反应等优点。在项目第一阶段，我们成功地(1)制造了10位数字的束珠，允许1,024(=210)个唯一代码，使其成为世界上复用程度最高的条形码磁珠；(2)设计并构建了微流控换能器和荧光检测器，用于逐一解码和检测荧光；(3)建立了P450(CYP2A7)、KCNA5和ITPase的多重PCR，以及珠子杂交；(4)利用多重PCR扩增序列对SNP基因分型方法进行了评估。第二阶段的工作将集中于临床应用的BEAM微珠平台的优化、BEAM系统的设计和集成、用于与药物反应相关的SNP基因分型的标准和定制SNP板的设计，以及用于靶基因扩增和SBE杂交的多重PCR的开发。

DESCRIPTION (provided by applicant): A Parallel High Throughput Pharmacogenetic Profiling Assay for Clinical Use Pharmacogenomic studies have already identified a number of genes whose SNP genotypes or haplotypes correlate with different individual drug responses, other metabolic processes or disease susceptibility. Thus the ability to determine genotypes quickly and accurately for medically relevant regions is critical to understanding the effects of an individual's genetic profile on these processes, and to the development of predictive, preventative and personalized medicine. Maxwell Sensors Inc. proposes to develop Barcode Etched Assayable Micro Bead (BEAM bead) technology, which combines digital barcoded beads and molecular chemistry for a pharmacogenetic SNP genotyping assay, which can be used for the high throughput molecular profiling of individuals. The digital BEAM beads are paramagnetic and are fabricated by photolithography to have thousands of identification codes and can be functionalized with nucleic acids, proteins or other small molecules to carry out large multiplexed assays in homogeneous or heterogeneous media. Genotyping using BEAM beads offers the advantages of flexibility, high throughput, easy fabrication, low cost mass production, and all in one reaction in small volumes. During Phase I of the project, we have successfully (1) fabricated 10 digit BEAM beads, allowing 1,024 (=210) unique codes, making it the most highly multiplexed barcode magnetic bead in the world; (2) designed and constructed a microfluidic transducer and fluorescence detector to decode and detect fluorescence one by one; (3) developed multiplex PCR for P450 (CYP2A7), KCNA5 and ITPase, and bead hybridization; and (4) evaluated the SNP genotyping assay using multiplex PCR amplified sequences. Phase II work will focus on optimization of the BEAM bead platform for clinical applications, design and integration of BEAM system, design of standard and custom SNP panels for genotyping SNPs relevant to drug responses, and development of multiplex PCR for target gene amplification and SBE hybridization.