Molecular Diagnostic Assays for Genetic Disease

遗传病的分子诊断分析

• 批准号: 6335572
• 负责人: Hashem Akhavan-Tafti
• 金额: $ 9.62万
• 依托单位: LUMIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2001-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6335572
• 关键词: coagulation factor V; diagnosis design /evaluation; gene mutation; genetic disorder diagnosis; genetic polymorphism; haptens; method development; nucleic acid amplification techniques; oligonucleotides

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.

描述（申请人的摘要）：第1阶段的目标是开发一种 检测人凝血因子V Leiden突变的灵敏试验和试剂盒 使用Lumigen发明的基于核酸连接的新技术。 连接技术是基于这样的发现： 寡核苷酸可以连续连接到模板结合的锚上 在其中短寡核苷酸不 与模板稳定杂交。缺乏稳定的杂交， 连接过程中的短寡核苷酸赋予极高的复制 忠诚在一些或所有短链上引入可检测的标签 寡核苷酸允许设计具有优异的生物活性的分子测定。 检测的特异性和可靠性。我们将使用一个小的半抗原标签， 用酶-抗体结合物和荧光酶检测 衬底 拟定商业应用： 拟议的检测方法和试剂盒应为开发一套 用于分析许多不同SNP的检测方法和产品。 这些测试包括 一种新的扩增方法，应比现有的扩增方法更具特异性 技术，同时实现可比的速度和灵敏度。 这些测试应该能够 在高通量平台上实现自动化，包括芯片格式。 这种方法的独特性质导致了一种 一种比PCR更特异的核酸扩增新技术， 生产更清洁的产品。我们将使用这种放大技术来增强 单核苷酸多态性检测方法的灵敏度 (SNP)与这种突变有关。开发一个单一的 用于区分该突变的所有三种可能基因型的测试将 也可以探索。这项测试将利用两种专利方法之一， 在Lumigen，使用专有技术检测两种酶标记分析物 荧光基质。