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: 7157209
• 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/7157209
• 关键词: biotechnology; blood tests; chemical cleavage; chemical stability; high throughput technology; magnetic field; method development; microprocessor /microchip; nucleic acid purification; particle; phosphorus compounds; polymerase chain reaction; reagent /indicator; surface coating

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（由申请人提供）：该II期项目继续开发和应用新材料，特别是磁性微粒，用于以极其快速的形式从细胞、组织和生物液体（包括全血）中直接分离DNA。这些材料消除了进行裂解步骤和从复杂材料中捕获DNA的需要，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专有化学，这使得它们能够在几乎任何生物条件下自发地和快速地裂解细胞并原位捕获暴露的DNA。因此，可以从全血中分离DNA而无需任何裂解步骤。独特的表面磷基团化学导致核酸以异常的强度结合，直到通过独特的切割机制释放，其中通过简单的化学反应选择性地破坏接头基团，以便从固体表面释放DNA。通过化学裂解而不是传统洗脱的释放防止了材料的过早、不受控制的释放并产生纯核酸。分离的DNA与下游应用如PCR兼容。正在探索另外的连接基团和裂解方法。第二阶段将发展专业知识来完成完整的合成，包括制备磁芯，包覆磁芯，以及用接头和头部基团进行功能化。将开发大规模制造功能化磁性颗粒的能力。我们已经生产出一种从全血中分离DNA的原型试剂盒。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，食品和培养基中的感染性微生物中捕获DNA，而无需进一步的分离步骤。这些材料有望在各种应用中简化样品制备和DNA提取。 第二阶段项目继续开发和应用新材料，特别是磁性微粒，用于以极其快速的方式直接从细胞，组织和生物液体（包括全血）中分离DNA。该材料消除了进行裂解步骤和从复杂基质中捕获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得到应用，其中DNA提取是必要的。