Innovative Molecular Platform for Prenatal Diagnostics

描述(由申请人提供)：本申请的目标是开发一种用于产前诊断的定制分子平台，该平台将我们专有的挂锁探头技术与下一代测序的能力相结合。分子平台比目前可用的方法更有效地将综合性与定制化结合在一起，并承诺将每个样品的商品成本降低到100美元以下。这项技术能够同时检测孟德尔疾病、非整倍体、单亲二体和不平衡结构异常。序列捕获在单个试管中与任何下一代测序平台相结合进行多路传输。由于这种方法很容易定制，分子遗传学家将能够为某些高危种族群体添加新的等位基因或创建定制的基因库。一旦它们被合成，单独的探头冷冻库就可以混合在一起，为几乎任何未来的应用创建定制的池。这项申请提出优化挂锁探头技术，然后建立一个20万个探针池用于产前诊断。首先，我们建议绘制产前专家感兴趣的18个孟德尔先天性疾病的疾病等位基因的基因组坐标。其次，我们建议建立一个1,000多个试点探针池，用于优化探针设计、合成和序列捕获。第三，我们建议使用这些优化的方法来合成一个专门用于产前诊断的200,000个复合探针池。最后，我们将在档案基因组DNA、羊水和胎儿有核红细胞(FNRBCs)上验证分子技术。最终，我们将把这项技术推向进行常规羊水筛查和非侵入性产前诊断的实验室。探针池还可用于任何常规的分子核型分析应用，例如癌症诊断或植入前遗传学诊断。 与公共卫生相关：我们正在开发新的方法，这些方法将有助于在婴儿出生前发现遗传问题。目前的方法都很昂贵，而且不能定制。我们正在使用DNA测序的新技术，有一天可能会让遗传学家更容易进行这一过程。

DESCRIPTION (provided by applicant): The goal of this application is to develop a customized molecular platform for prenatal diagnosis that combines our proprietary padlock probe technology with the power of next-generation sequencing. The molecular platform combines comprehensiveness with customization more effectively than currently available methods, and promises to reduce per sample cost of goods to below $100. The technology enables simultaneous detection of Mendelian disorders, aneuploidy, uniparental disomy, and unbalanced structural abnormalities. Sequence capture is performed multiplexed in a single tube, in conjunction with any next-generation sequencing platform. Because the method is easy to customize, molecular geneticists will be able to add new alleles or create customized pools for certain at-risk ethnic populations. Once they are synthesized, individual probe freezer stocks can be mixed to create customized pools for virtually any future application. This application proposes to optimize the padlock probe technology and then build a 200,000-plex probe pool for prenatal diagnostics. First, we propose to map the genome coordinates for disease alleles from 18 Mendelian congenital disorders of interest to the prenatal specialist. Second, we propose to build a 1,000-plex pilot probe pool that will be used to optimize probe design, synthesis, and sequence capture. Third, we propose to use these optimized to synthesize a 200,000-plex probe pool specifically for prenatal diagnostics. Finally, we will validate the molecular technology on archival genomic DNA, amniotic fluid, and fetal nucleated red blood cells (FNRBCs). Eventually, we will market the technology to laboratories performing routine amniotic screening as well as noninvasive prenatal diagnostics. The probe pool could also be used generically for any routine molecular karyotyping application, such as cancer diagnostics or preimplantation genetic diagnosis. PUBLIC HEALTH RELEVANCE: We are developing new methods that will help to detect genetic problems before babies are born. Current methods are expensive and not customizable. We are using new technology for DNA sequencing that may one day make this process easier for geneticists.