Molecular Diagnostic Assays for Genetic Disease

描述(申请人摘要)：第一阶段的目标是开发一个 检测人凝血因子V Leiden突变的敏感试验及试剂盒 血液使用了卢米根发明的一种基于核酸连接的新技术。 结扎技术是基于一项发现，即大量短链 寡核苷酸可以连续连接到模板结合的锚定上 在短寡核苷酸不存在的条件下进行的序列 稳定地与模板杂交。没有稳定的杂交 在连接过程中的短寡核苷酸可提供极高的复制能力 富达。在一些或全部短片上加入可检测的标签 寡核苷酸允许设计具有优异性能的分子分析方法 检测的特异性和可靠性。我们将使用一个小的半抗原标签来 用酶-抗体结合物和化学发光酶进行检测 底物。 建议的商业应用： 建议的测试方法和工具包应该为开发一套 用于分析多种不同SNPs的检测方法和产品。这些测试涉及一个 新的扩增过程应该比现有的扩增更具体 技术，同时获得与之相当的速度和灵敏度。测试应该能够 在包括芯片格式的高吞吐量平台上实现自动化。 这种方法的独特性质导致了一种 比聚合酶链式反应和聚合酶链式反应更特异的核酸扩增新技术 生产更清洁的产品。我们将利用这种放大技术来增强 单核苷酸多态检测方法的敏感性 (SNP)与该突变相关。开发一种单一的 对这一突变的所有三种可能的基因类型的区分测试将 也是被探索的。这项测试将使用开发的两种专利方法之一 在Lumigen使用专利技术检测两种酶标记分析物 化学发光衬底。

DESCRIPTION (Applicant's Abstract): The objective of Phase 1 is to develop a sensitive test and kit for detection of the Factor V Leiden mutation in human blood using a new nucleic acid ligation-based technology invented at Lumigen. The ligation technology is based on the discovery that a multitude of short oligonucleotides can be contiguously ligated to a template-bound anchor sequence performed under conditions in which the short oligonucleotides do not stably hybridize to the template. The absence of stable hybridization of the short oligonucleotides during the ligation confers extremely high replication fidelity. Incorporation of detectable labels on some or all of the short oligonucleotides permits the design of molecular assays with excellent specificity and reliability of detection. We will use a small hapten label to be detected with an enzyme-antibody conjugate and a chemiluminescent enzyme substrate. PROPOSED COMMERCIAL APPLICATION: The proposed test methods and kits should provide the basis for developing a set of detection methods and products for analysis of many different SNPs. The tests involve a new amplification process which should be more specific than existing amplification technology while achieving comparable speed and sensitivity. The tests should be capable of automation on a high throughput platform including chip formats. The unique properties of this methodology have resulted in the creation of a new nucleic acid amplification technology which is more specific than PCR and yields cleaner products. We will use this amplification technique to enhance the sensitivity of the method to detect the single-nucleotide polymorphism (SNP) associated with this mutation. The feasibility of developing a single test for discrimination of all three possible genotypes of this mutation will also be explored. This test will utilize one of two patented methods developed at Lumigen for detection of two enzyme labeled analytes using proprietary chemiluminescent substrates.