Development of Liposome Complexed RNA Quality Controls

描述(申请人提供)：新的分子实验室测试的爆炸性增长极大地改善了白血病的诊断和管理，但用于测试验证的可再生稳定对照的问题尚未解决。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，可能具有传染性，供应有限，无法通过常规测试提取进行处理。患者的保密性是有问题的，每个样本只有一个基因，因此控制包括基因芯片在内的多重测试的成本高得令人望而却步。第二阶段将生产合成的、稳定的、非传染性的、多分析物控制，以保证基于分子的白血病测试的质量。我们的新方法的可行性通过第一阶段生产稳定的阳离子脂质体复合RNA来证明，可用作BCR-ABL临床试验的对照。第二阶段制造全面的白血病控制系统，首先创建另外七个包含白血病基因片段的克隆库。RNA将在体外生产，并用第一阶段专利技术产生的脂质体复合体稳定下来。参考基因片段将被纳入，以使对照对用于指导治疗的最小残留疾病量化测试有用。该项目为支持和实施白血病的关键分子检测做出了重大贡献。

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.