Ultra-Rare Circulating Tumor DNA Detection with PAP & Digital Bead Technologies

使用 PAP 检测超罕见循环肿瘤 DNA

• 批准号: 7348452
• 负责人: Winston Z. Ho
• 金额: $ 21万
• 依托单位: MAXWELL SENSORS, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7348452
• 关键词: 2' ,3' -Dideoxyadenosine; Alleles; Automation; Bar Codes; Biological Markers; Blood; Cancer Diagnostics; Cancer Prognosis; Cities; Clinical; Coupled; DNA; DNA Markers; DNA-Directed DNA Polymerase; Detection; Diagnosis; Diagnostic; Diphosphates; Epigenetic Process; Excision; Figs - dietary; Frequencies; Genes; Genetic; Genomics; Genotype; In Vitro; Invasive; Left; Malignant Neoplasms; Medical center; Microfluidics; Microspheres; Molecular; Mutate; Mutation; Nucleotides; Numbers; Oligonucleotide Primers; Oligonucleotides; Patient Monitoring; Patients; Phase; Play; Preventive; Primary Neoplasm; Property; Protein p53; Proteins; Resolution; Right-On; Role; Scanning; Screening for cancer; Solid Neoplasm; Somatic Mutation; System; Systems Integration; TP53 gene; Technology; Testing; Thick; Time; Tumor Suppressor Genes; Work; base; cancer therapy; cost; design; digital; gel electrophoresis; interest; mutant; peripheral blood; polymerization; prognostic; prototype; sensor; tumor

DESCRIPTION (provided by applicant): Diagnosis and prognosis of cancer have increasingly benefited from the use of molecular markers and molecular approaches. Accumulating lines of evidence have shown that there is tumor DNA circulating in the peripheral blood of patients with a variety of cancers. Multiple studies have shown that circulating DNA carries genetic and epigenetic alterations that are found in the corresponding primary tumors. Therefore, the presence of circulating DNA in blood is a strong diagnostic and prognostic indicator, making blood a valuable solid tumor surrogate for non-invasive and preventive cancer diagnostics as well as monitoring of patients who are actively under cancer therapy. One of the most commonly used targets for detecting tumor DNA in blood are p53 mutations. The p53 tumor suppressor gene product (TP53) is playing an important role in early cancer initiation. Maxwell Sensors Inc. together with City of Hope National Medical Center proposes to develop a "TP53-TYPER" system for early cancer screening. The prototype TP53-TYPER will be designed to (i) amplify ultra-rare TP53 mutations within the excess of normal genomic DNA circulating in blood, (ii) capture the amplified alleles onto digitally bar-coded beads by hybridization, and (iii) identify, detect and quantify the amplified alleles. The technology combining Bi-directional PAP allele specific amplification (Bi-PAP) with the capture of Bi-PAP amplified TP53 somatic mutations on digital microbeads along with identification and fluorescent scanning of allele abundance, we are enabling high-throughput and high resolution genotyping to be performed in a reliable, cost-effective and time-saving manner. The combination of Bi-PAP, an amplification technology with ultra-high sensitivity (1 mutant DNA molecule) and selectivity (1 mutant in 109 to 1011 wild-type molecules), with addressable bead-based microfluidic detection will enable the routine use of biomarkers in blood in early clinical cancer diagnostics. The TP53-TYPER system is amenable to commercial system integration and automation for efficient large scale analyses. The Phase I work will focus on system fabrication and feasibility testing.

描述（由申请人提供）：癌症的诊断和预后越来越受益于分子标记和分子方法的使用。越来越多的证据表明，多种癌症患者的外周血中都存在肿瘤DNA循环。多项研究表明，循环 DNA 携带在相应原发性肿瘤中发现的遗传和表观遗传改变。因此，血液中循环 DNA 的存在是一个强有力的诊断和预后指标，使血液成为非侵入性和预防性癌症诊断以及积极接受癌症治疗的患者监测的有价值的实体瘤替代物。 p53 突变是检测血液中肿瘤 DNA 最常用的靶标之一。 p53 肿瘤抑制基因产物 (TP53) 在早期癌症发生中发挥着重要作用。 Maxwell Sensors Inc. 与希望之城国家医疗中心共同提议开发一种用于早期癌症筛查的“TP53-TYPER”系统。原型 TP53-TYPER 的设计目的是 (i) 扩增血液中循环的过量正常基因组 DNA 中的极其罕见的 TP53 突变，(ii) 通过杂交将扩增的等位基因捕获到数字条形码珠上，以及 (iii) 识别，检测并量化扩增的等位基因。该技术将双向 PAP 等位基因特异性扩增 (Bi-PAP) 与数字微珠上 Bi-PAP 扩增的 TP53 体细胞突变的捕获以及等位基因丰度的识别和荧光扫描相结合，我们正在实现高通量和高分辨率基因分型以可靠、经济高效且节省时间的方式进行。 Bi-PAP 是一种具有超高灵敏度（1 个突变 DNA 分子）和选择性（109 至 1011 个野生型分子中的 1 个突变）的扩增技术，与可寻址珠基微流体检测相结合，将使生物标记物的常规使用成为可能早期临床癌症诊断中的血液中。 TP53-TYPER 系统适合商业系统集成和自动化，以进行高效的大规模分析。第一阶段的工作将侧重于系统制造和可行性测试。