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（下一代测序）的作用，人们讨论已久。 与实时PCR相比，在临床诊断、预后和筛查中。NGS提供无与伦比的 发现能力并代表了最终的多路复用能力。然而，NGS 成本高昂，并且通常需要复杂的工作流程，周转时间较长。实时PCR，在 相比之下，相对便宜，并且可以完全自动化，以便在 护理与快速样品在结果出来的时间。但实时PCR只能检测已知序列 并且具有非常有限的多路复用能力（通常为4-6色）。 在临床诊断、预后、甚至筛查环境中，查询一个患者的年龄通常是足够的。 已知的基因组或突变。当需要有限的多路复用时，例如4或5种颜色 在单一反应中，这可能是伴随诊断的情况，实时PCR已经被应用于 非常成功的部署。但目前，实时PCR要想渗透比较困难 诊断或筛选测定，其需要具有数十或数百范围大小的板。为 尽管NGS的工作流程更复杂，成本更高， 周转时间 为了解决上述未满足的临床诊断需求，该项目旨在开发 一种实现高度多重PCR检测的新方法，用于实时PCR和数字PCR。 PCR法鉴于PCR在分子诊断中的重要作用， 完成后，该项目预计将大大扩大实用性和可访问性的高度 多重PCR检测在疾病诊断和筛查中的应用。