Significant expansion of spectral multiplexing in PCR

PCR 中光谱多重功能的显着扩展

• 批准号: 10698894
• 负责人: Jiangbo Yu
• 金额: $ 49.99万
• 依托单位: LAMPROGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698894
• 关键词: Achievement; Address; Base Sequence; Bioinformatics; Biological Assay; Clinical; Clinical Medicine; Color; Compensation; Complex; Computer software; Custom; DNA Sequence; Development; Diagnostic; Disease; Dyes; Early Diagnosis; Energy Transfer; Exhibits; Family; Flow Cytometry; Fluorescence; Fluorescent Dyes; Fluorescent Probes; Future; Gene Expression Profiling; Genes; Genetic; Genomics; Genotype; Hemorrhage; Hydrophobicity; Infectious Agent; Licensing; Measurement; Measures; Methods; Mutation; Nature; Oligonucleotides; Outcome; Penetration; Performance; Pharmacotherapy; Play; Polymers; Prognosis; Proteins; Protocols documentation; Public Health; Quantum Dots; Reaction; Reproducibility; Risk; Role; Running; Sampling; Science; Screening for cancer; Series; System; Technology; Time; Tube; Validation; Western Blotting; carrier testing; clinical diagnosis; clinical diagnostics; companion diagnostics; comparative; cost; design; diagnostic assay; digital; dimer; disease diagnosis; improved; innovation; instrument; liquid biopsy; method development; molecular diagnostics; next generation sequencing; novel strategies; nucleic acid quantitation; operation; personalized medicine; point of care; prognostic; reproductive; screening

Project Summary There have been long-standing discussions about the role of NGS (next-generation sequencing) versus real-time PCR in clinical diagnosis, prognosis, and screening. NGS offers unrivaled discovery power and represents the ultimate in multiplexing capability. NGS, however, is expensive and often requires complex workflows with a long turnaround time. Real-time PCR, in contrast, is comparatively inexpensive and can be fully automated for operation at the point-of- care with a fast sample-in-result-out time. But real-time PCR can only detect known sequences and has a very limited multiplexing capability (typically 4-6 colors). In a clinical diagnostic, prognostic, and even screening setting, it is often sufficient to query a known panel of genes or mutations. When limited multiplexing is required, such as 4 or 5 colors in a single reaction, which can be the case in companion diagnostics, real-time PCR has been very successfully deployed. But at present, it is more difficult for real-time PCR to penetrate diagnostic or screening assays that require panels with sizes in the tens or hundreds range. For the larger panels, NGS is often used despite its more complex workflow, higher cost, and longer turnaround time. To address the above described unmet need in clinical diagnostics, this project aims to develop a new approach to achieve highly multiplexed PCR assays, for both real-time PCR and digital PCR. Given the important role PCR plays in molecular diagnostics, when successfully completed, this project is expected to greatly expand the utility and accessibility of highly multiplexed PCR assays in disease diagnosis and screening.

项目摘要 关于NGS的作用（下一代测序）的长期讨论 与实时PCR相对于临床诊断，预后和筛查。 NGS提供无与伦比的 发现能力，代表了多路复用能力的最终功能。但是，NGS是 昂贵的，通常需要漫长的周转时间复杂的工作流程。实时PCR，在 对比，相对便宜，可以在 - 可以完全自动化以进行操作 用快速的样本淘汰时间来照顾。但是实时PCR只能检测到已知序列 并且具有非常有限的多路复用能力（通常为4-6种颜色）。 在临床诊断，预后甚至筛查设置中，通常足以查询 已知的基因或突变面板。如果需要有限的多路复用，例如4或5种颜色 在单一反应中，在伴侣诊断中可能是这种情况，实时PCR一直是 非常成功地部署。但是目前，实时PCR很难穿透 诊断或筛选测定法需要在数十个或数百个范围内具有尺寸的面板。为了 尽管工作流程更复杂，成本更高，并且更长 周转时间。 为了满足上述临床诊断中所述未满足的需求，该项目旨在发展 用于实时PCR和数字的新方法来实现高度多重的PCR分析 pcr。鉴于PCR在分子诊断中的重要作用，当成功 完成后，该项目有望大大扩大高度的实用性和可访问性 疾病诊断和筛查中的多重PCR分析。