A novel instrument for high efficiency extraction of circulating nucleic acids

一种高效提取循环核酸的新型仪器

• 批准号: 7387763
• 负责人: ANDRE MARZIALI
• 金额: $ 12.15万
• 依托单位: UNIVERSITY OF BRITISH COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-07 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7387763
• 关键词: Acrylamide; Acrylamides; Applications Grants; Automation; Award; Biological Markers; Biotechnology; Blood Stains; Body Fluids; Cells; Clinical; Collection; Communities; Complex; DNA; DNA Markers; DNA Sequence; Detection; Development; Feces; Forensic Medicine; Future; Gel; Genomics; Human; Licensing; Link; Malignant Neoplasms; Metagenomics; Methods; Modification; Mutation; Non-Invasive Cancer Detection; Nucleic Acids; Oncogene Activation; Performance; Petroleum; Process; Range; Recovery; Research; Risk; Sampling; Science; Sepharose; Serum; Signal Transduction; Silicon Dioxide; Site; Soil; Specificity; Tars; Technology; Testing; UBC gene; Urine; Validation; Work; base; commercialization; concept; cost; desire; electric field; infancy; innovation; instrument; microchip; neoplastic cell; novel; tool; tumor

DESCRIPTION (provided by applicant): Circulating nucleic acids from dying tumor cells may yield an early indication of cancer if they could be detected at sufficiently low abundance in body fluids in the presence of background DNA from healthy cells as well as contaminants inherent to blood serum, urine, and stool samples. Methods capable of enriching for small quantities of partially intact tumor DNA from body fluids while rejecting contaminants and background DNA from healthy cells would be a great advantage. This would be particularly true, if the DNA could be enriched for sequence mutations that signal oncogene activation or other tumor related sequence signatures. We have developed a method of DNA concentration and extraction (SCODA) that, in its simplest form, promises to increase the amount and purity of DNA extracted from body fluids, while reducing the amount of processing steps and cost. SCODA will allow facile extraction of nucleic acid biomarkers from samples such as urine, stool, or blood serum for non-invasive cancer detection, and for genomic studies of cancer. SCODA provides a means for high efficiency concentration of low abundance DNA with a high degree of contaminant rejection. Applications of this technology to forensics and genomics research are being commercialized, and its value has been proven in both fields. We aim to apply SCODA technology to extraction of circulating DNA from body fluids ranging from blood serum to stool. We expect that its outstanding efficiency and rejection of contaminants, as proven in our forensics work, will yield benefits in biomarker extractions. Furthermore, we plan to make modifications to SCODA that will allow enrichment for specific low abundance DNA sequences. This will be done by altering the concentration mechanism, such that only DNA fragments containing specific sequences relating to tumor cells will be concentrated. This promises to provide a pre-concentration tool that would enable exceptionally sensitive detection of tumor DNA biomarkers in body fluids.

描述（由申请人提供）：如果可以在健康细胞的背景DNA以及血清，尿液，尿液和粪便固有的污染物的情况下，可以在体液中足够低的体液中检测到足够低的体液，则可以在体液的足够低度中检测到癌症的早期指示。能够从体液中富集少量部分完整的肿瘤DNA的方法，而拒绝健康细胞的污染物和背景DNA将是一个很大的优势。如果可以将DNA富含信号过癌基因激活或其他相关肿瘤相关序列特征的序列突变，则尤其如此。 我们已经开发了一种DNA浓度和提取方法（SCODA）的方法，该方法以最简单的形式有望增加从体液中提取的DNA的量和纯度，同时减少处理步骤和成本的量。 SCODA将允许从尿液，粪便或血清等样品中轻松提取核酸生物标志物，以进行非侵入性癌症检测，并用于癌症的基因组研究。 SCODA提供了一种高效率浓度的低丰度DNA，并具有高度污染物排斥。该技术在法医和基因组学研究中的应用正在商业化，其价值在这两个领域都得到了证明。 我们的目标是将SCODA技术应用于从血清到凳子的体液中提取循环DNA。我们预计，正如我们的取证工作中所证明的那样，其出色的效率和拒绝污染物将在生物标志物提取方面产生好处。此外，我们计划对SCODA进行修改，以允许对特定的低丰度DNA序列富集。这将通过改变浓度机制来完成，以便将仅包含与肿瘤细胞有关的特定序列的DNA片段浓缩。这有望提供一种预浓缩工具，该工具将使体液中肿瘤DNA生物标志物具有异常敏感的检测。