Development of Liposome Complexed RNA Quality Controls

脂质体复合 RNA 质量控制的开发

• 批准号: 6642059
• 负责人: Clark A. Rundell
• 金额: $ 34.18万
• 依托单位: MAINE MOLECULAR QUALITY CONTROLS, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6642059
• 关键词: RNA; analytical method; biomedical equipment development; diagnosis quality /standard; liposomes; microarray technology; molecular cloning; polymerase chain reaction; synthetic nucleic acid; technology /technique development; transfection

DESCRIPTION (provided by applicant): The explosion of new molecular laboratory tests has significantly improved diagnosis and management of leukemia, but the problem of a renewable supply of stable controls for test validation has not been resolved. The current paradigm of using cell lines or patient specimens for quality assurance is unsatisfactory because materials are unstable, potentially infectious, of limited supply, and cannot be processed through routine test extractions. Patient confidentiality is problematic and each specimen has only one genotype, thus making control of multiplex tests, including gene chips, prohibitively expensive. Phase II will produce synthetic, stable, non-infectious, multi-analyte controls for quality assurance of molecular-based leukemia tests. Feasibility of our novel approach was demonstrated by Phase I production of stable, cationic liposome-complexed RNA, useful as controls in BCR-ABL clinical tests. Phase II manufacture of a comprehensive Leukemia Control Set begins by creating seven additional clone banks containing leukemic gene segments. RNA will be produced in vitro and stabilized with liposome complexes generated by Phase I proprietary techniques. Reference gene segments will be incorporated to render controls useful for minimal residual disease quantification tests used to direct therapies. The proposed project makes a significant contribution to the support and implementation of critical molecular tests for leukemia.

描述（由申请人提供）：新的分子实验室测试的激增显着改善了白血病的诊断和管理，但用于测试验证的稳定对照的可再生供应问题尚未解决。目前使用细胞系或患者样本进行质量保证的范例并不令人满意，因为材料不稳定、具有潜在传染性、供应有限，并且无法通过常规测试提取进行处理。患者保密性存在问题，并且每个样本只有一种基因型，因此使得包括基因芯片在内的多重测试的控制成本过高。第二阶段将生产合成的、稳定的、非传染性的多分析物对照品，以保证基于分子的白血病检测的质量。我们的新方法的可行性通过稳定的阳离子脂质体复合 RNA 的 I 期生产得到了证明，该 RNA 可用作 BCR-ABL 临床测试中的对照。全面的白血病控制组的第二阶段生产首先创建七个包含白血病基因片段的额外克隆库。 RNA 将在体外生产，并通过第一阶段专有技术产生的脂质体复合物进行稳定。将纳入参考基因片段，以提供可用于指导治疗的微小残留疾病量化测试的对照。拟议的项目为支持和实施白血病关键分子检测做出了重大贡献。