Lab-in-a-Tube Technology for Genotyping at Point-of-Care

用于床旁基因分型的试管实验室技术

• 批准号: 6834378
• 负责人: BERTRAND LEMIEUX
• 金额: $ 15.41万
• 依托单位: IQUUM, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-30 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6834378
• 关键词: biomarker; biotechnology; clinical research; diagnosis design /evaluation; fluorescent dye /probe; genotype; high throughput technology; human subject; nucleic acid hybridization; phase change; polymerase chain reaction; single nucleotide polymorphism; technology /technique development

DESCRIPTION (provided by applicant): The goal of this project is to develop a high-level multiplex real time polymerase chain reaction (PCR) assay for single nucleotide polymorphisms (SNP) using IQuum's innovative Lab-in-a-tube (Liat TM) technology. Liat technology is capable of extracting genomic DNA from whole blood and detecting sequences, using real time PCR, in 30 minutes. Liat technology is ideal for the point of care because it reduces the complexity of nucleic acid tests to a minimum. Indeed, the operator of the Liat Molecular Analyzer, the instrument that performs this genomic DNA sequence detection assay, is only required to input blood into the Liar Tubule and insert the closed Tubule into the instrument. The Liat Molecular Analyzer can also perform multi-color fluorescent probe melting assays in 8 minutes. In Phase I, we propose to investigate the applicability of the molecular inversion probe assay to real time PCR detection using multi-color fluorescent probe melting as a means of detecting multiple SNPs. In addition, we propose to develop a set of hybridization tags with defined melting profiles in order to maximize the multiplexing of this assay. Traditional multiplex real time PCR can only accommodate up to 2 SNPs in one assay. Our strategy will allow us to score up to 40 SNPs in a single assay. We estimate this order of magnitude increase in multiplexing will be accomplished with a 1 hour assay that can be deployed at the point-of-care. In Phase II, we plan to develop a multi-tube Liat Molecular Analyzer that will have the capability of scoring 1.5 million genotypes per year, at the point-of-care. In addition, we propose to develop assays for scoring 40 SNPs that have been associated with increased risk of arterial thrombosis. Venous thrombosis occurs in 0.1 percent of the population each year, and its most severe complication (pulmonary embolism) is as frequent as strokes in developed countries. Arterial thrombosis is a key component of myocardial infarction, stroke and peripheral artery disease, all of which account for >19 million deaths annually. In Phase III, we would collaborate with researchers at Harvard Medical School and Tufts Medical School on clinical trials to evaluate the risk associated with the markers.

描述（由申请人提供）：该项目的目的是使用IQuum的创新性实验室（LIAT TM）技术开发单核苷酸多态性（SNP）的高级多重实时聚合酶链反应（PCR）测定。 LIAT技术能够在30分钟内使用实时PCR从全血和检测序列中提取基因组DNA。 LIAT技术是护理点的理想选择，因为它将核酸测试的复杂性降低到最低。实际上，只需要将血液输入骗子小管中，然后将封闭的小管插入仪器中，则需要使用该基因组DNA序列检测测定法的仪器（执行此基因组DNA序列检测测定法）的操作员。 LIAT分子分析仪还可以在8分钟内执行多色荧光探针融化测定。在第一阶段，我们建议使用多色荧光探针融化作为检测多个SNP的手段，研究分子反转探针测定对实时PCR检测的适用性。此外，我们建议开发一组具有定义熔解曲线的杂交标签，以最大程度地提高此测定的多路复用。传统的多重实时PCR只能在一个测定中最多可容纳2个SNP。我们的策略将使我们在一个测定中得分多达40个SNP。我们估计，将通过1小时的测定法完成多重综述的数量级增加。在第二阶段，我们计划开发一个多管LIAT分子分析仪，该分子分析仪将在护理点每年获得150万个基因型的能力。此外，我们建议开发与已增加动脉血栓形成风险增加的40个SNP的测定法。静脉血栓形成每年发生在0.1％的人口中，其最严重的并发症（肺栓塞）与发达国家中风一样频繁。动脉血栓形成是心肌梗塞，中风和周围动脉疾病的关键组成部分，所有这些都占每年> 1900万人死亡。在第三阶段，我们将与哈佛医学院和塔夫茨医学院的研究人员进行临床试验，以评估与标记相关的风险。