Materials for Isolation of Nucleic Acids from Whole Blood without a Lysis Step

无需裂解步骤即可从全血中分离核酸的材料

• 批准号: 7265240
• 负责人: Hashem Akhavan-Tafti
• 金额: $ 36.57万
• 依托单位: LUMIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7265240
• 关键词: Address; Anticoagulants; Area; Bacteria; Binding; Biological; Biological Sciences; Blood; Cells; Chemistry; Class; Cleaved cell; Compatible; Complex; Condition; Confidentiality; Culture Media; Cytolysis; DNA; DNA Viruses; Development; Diagnostic; Evaluation; Feedback; Goals; Head; Housing; In Situ; Industry; Link; Liquid substance; Magnetism; Methods; Nucleic Acid Binding; Nucleic Acids; Numbers; Performance; Phase; Plasmids; Polymerase Chain Reaction; Preparation; Property; Protocols documentation; Range; Reporting; Research; Research Design; Sampling; Solid; Spontaneous Rupture; Surface; Technology; Testing; Time; Tissues; Urine; Whole Blood; Work; base; cell type; chemical cleavage; chemical reaction; commercialization; improved; microorganism; particle; plasmid DNA; prevent; prototype; scale up

DESCRIPTION (provided by applicant): This Phase II project continues the development and application of new materials, especially magnetic microparticles for directly isolating DNA from cells, tissues, and biological fluids, including whole blood, in an extremely rapid format. The materials eliminate the need to perform a lysis step and capture DNA from complex materials thereby simplifying sample preparation methods. The functionalized microparticles for binding and releasing nucleic acids utilize a proprietary chemistry which enables them to spontaneously and rapidly lyse cells and capture the exposed DNA in situ under virtually any biological conditions. As a result DNA can be isolated from whole blood without any lysis step. The unique surface phosphonium group chemistry causes nucleic acids to bind with exceptional strength until released by a unique cleavage mechanism where a linker group is selectively broken by a simple chemical reaction in order to free DNA from the solid surface. Release by chemical cleavage rather than traditional elution prevents premature, uncontrolled release of material and yields pure nucleic acid. Isolated DNA is compatible with downstream applications such as PCR. Additional linking groups and cleavage methods are being explored. Phase II will develop the expertise to perform the complete synthesis including preparing the magnetic cores, coating the cores, and functionalizing with linker and head groups. The capability to manufacture the fuctionalized magnetic particles on a large scale will be developed. We have produced a prototype kit for isolating DNA from whole blood. We areing the range of applications of these new materials is being expanded to plasmid purification, isolation of DNA from viruses, and capture of DNA from infectious microorganisms in urine, foodstuffs and culture media with no further isolation steps. The materials are expected to streamline sample preparation and DNA extraction across a broad variety of applications. This Phase II project continues the development and application of new materials, especially magnetic microparticles for directly isolating DNA from cells, tissues, and biological fluids, including whole blood, in an extremely rapid format. The materials eliminate the need to perform a lysis step and capture DNA from complex matrices thereby simplifying sample preparation methods. The materials will find use in a great number of applications in the life sciences and diagnostics industries for which DNA extraction is necessary.

描述(申请人提供)：该第二阶段项目继续开发和应用新材料，特别是磁性微粒，以极快的方式直接从细胞、组织和生物液(包括全血)中分离DNA。这种材料不需要执行裂解步骤并从复杂材料中捕获DNA，从而简化了样品制备方法。用于结合和释放核酸的功能化微粒利用一种专利化学物质，使它们能够自发和快速地裂解细胞，并在几乎任何生物条件下原位捕获暴露的DNA。因此，不需要任何裂解步骤，就可以从全血中提取DNA。独特的表面膦基团化学使核酸以非凡的强度结合，直到通过独特的切割机制释放，其中连接基通过简单的化学反应选择性地断裂，以便将DNA从固体表面释放出来。通过化学裂解释放，而不是传统的洗脱，可以防止物质过早、不受控制地释放，并产生纯核酸。分离的DNA与下游应用，如聚合酶链式反应，是兼容的。正在探索其他连接基团和裂解方法。第二阶段将开发进行完整合成的专业知识，包括制备磁芯、涂覆磁芯以及使用连接基和磁头基进行功能化。将发展大规模制造功能化磁性颗粒的能力。我们已经生产了一种从全血中提取DNA的原型试剂盒。我们知道，这些新材料的应用范围正在扩大到质粒纯化，从病毒中分离DNA，以及从尿液、食品和培养基中捕获感染微生物的DNA，而不需要进一步的分离步骤。这些材料有望在广泛的应用中简化样品准备和DNA提取。这一二期项目继续开发和应用新材料，特别是磁性微粒，以极快的方式直接从细胞、组织和生物液(包括全血)中分离DNA。这种材料消除了执行裂解步骤并从复杂基质中捕获DNA的需要，从而简化了样品制备方法。这些材料将在生命科学和诊断行业的大量应用中找到用途，而这些行业的DNA提取是必要的。