Multiplex Detection of Mutations in Myeloproliferative Disorder and Leukemia Pati

骨髓增殖性疾病和白血病突变的多重检测

• 批准号: 7326750
• 负责人: FEI YE
• 金额: $ 25.6万
• 依托单位: AMBION DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7326750
• 关键词: Acute Myelocytic Leukemia; Adult Acute Myeloblastic Leukemia; Age; Alleles; American Cancer Society; Biological Assay; Blood specimen; Cause of Death; Cell Line; Child; Chronic; Chronic Myeloid Leukemia; Classification; Clinical; Clinical Management; Complex; Costs and Benefits; Cytogenetics; DNA; Detection; Diagnosis; Diagnostic; Disease; Frequencies; Gene Mutation; Genes; Genetic; Goals; Hemorrhagic Thrombocythemia; Hour; Human; In Vitro; Janus kinase 2; KIT gene; Karyotype; Laboratories; Lesion; Liquid substance; Malignant Neoplasms; Molecular; Mutation; Myeloid Leukemia; Myeloproliferative disease; NPM1 gene; Numbers; Outcome; Pathogenesis; Patients; Phase; Physicians; Point Mutation; Polycythemia Vera; Polymerase Chain Reaction; Primary Myelofibrosis; RNA; Reagent; Recurrence; Reporting; Sampling; Screening procedure; Signal Pathway; Subgroup; System; Technology; Testing; Therapeutic; Transcript; Tube; Tyrosine Kinase Inhibitor; United States Food and Drug Administration; base; chemotherapy; clinically relevant; cost; design; disease classification; human MPL protein; improved; inhibitor/antagonist; leukemia; multiplex detection; nucleophosmin; outcome forecast; prognostic; rapid detection; research clinical testing; software development; young adult

DESCRIPTION (provided by applicant): The overall objective for this project is to develop a diagnostic assay for rapid detection of major mutations in patients with acute myeloid leukemia (AML), non-BCR/ABL chronic myelogenous leukemia (CML) and myeloproliferative disorders (MPDs). We propose to develop a single tube multiplex assay which uses high fidelity PCR for amplification of target mutations and hybridization of Luminex bead-tagged probes for detection of specific sequences. We have successfully developed highly multiplexed DNA- and RNA-based assays using this technology. The detection of point mutations in a background of wild-type transcripts will provide substantial benefits for diagnostic and prognostic testing and clinical management of these diseases. According to the American Cancer Society, 34,800 new cases of all types of leukemia were reported in the US in 2005. Leukemia is the number one cause of deaths from cancer in children and young adults under age 20. Although myeloid malignancies have been traditionally classified by their morphological and cytogenetic features, mutations that are undetectable at a chromosomal level have been shown to be correlated with distinct prognosis in leukemia and MPDs. As the molecular pathogenesis for these diseases is becoming evident, and mutation-specific inhibitors become available, classification will need to include molecular identification. It is our goal to develop a comprehensive assay system which 1) rapidly detects aberrant sequences for resolving ambiguous diagnosis of AML, non-BCR/ABL CML and MPDs, and 2) is predictive of therapeutic outcomes and thus useful in selecting between different treatment options. The availability of this assay for leukemia and MPD mutations should greatly improve the clinical decisions necessary for effective treatment of leukemia and MPD patients while improving clinical laboratory workflow and reducing costs. The Specific Aims are: 1) Develop a rapid single-tube assay for the simultaneous detection of major mutations in patients with AML and CML/MPDs. 2) Develop comprehensive controls for the clinical evaluation of the assay. In Phase II, we will evaluate the diagnostic value of the assay against a large number of clinical samples in a diagnostic laboratory setting. We will develop software for analyzing the assay results and to assist in clinical interpretation of the results. The final outcome is to launch mutation-specific Analyte Specific Reagents (primers, probes) and apply for FDA approval as an In Vitro Diagnostic Assay. The availability of this assay for leukemia and MPD mutations should greatly improve the clinical decisions necessary for effective treatment of leukemia and MPD patients while improving clinical laboratory workflow and reducing costs. The overall goal of this proposal is to develop an assay to detect mutations that are important for determining treatment of myeloproliferative disorders and leukemia. This liquid bead array format of this assay will provide efficiency and cost benefits to diagnostic laboratories and provide crucial information to physicians and patients for use in making treatment decisions.

描述（由申请人提供）：本项目的总体目标是开发一种快速检测急性髓性白血病（AML）、非BCR/ABL慢性髓性白血病（CML）和骨髓增生性疾病（MPD）患者主要突变的诊断方法。我们建议开发一种单管多重检测方法，该方法使用高保真PCR扩增靶突变和Luminex珠标记探针杂交检测特定序列。我们已经成功地开发了高度多路复用的DNA和RNA为基础的分析使用这项技术。在野生型转录本的背景下检测点突变将为这些疾病的诊断和预后测试以及临床管理提供实质性益处。根据美国癌症协会的数据，2005年美国报告了34,800例各种类型的白血病新病例。白血病是儿童和20岁以下年轻人死于癌症的头号原因。尽管骨髓恶性肿瘤传统上是根据其形态学和细胞遗传学特征进行分类的，但在染色体水平上检测不到的突变已被证明与白血病和MPD的不同预后相关。随着这些疾病的分子发病机制越来越明显，突变特异性抑制剂变得可用，分类将需要包括分子鉴定。我们的目标是开发一种全面的检测系统，该系统1）快速检测异常序列，以解决AML、非BCR/ABL CML和MPD的模糊诊断，2）预测治疗结果，因此可用于在不同治疗方案之间进行选择。这种用于白血病和MPD突变的检测方法的可用性将大大改善有效治疗白血病和MPD患者所需的临床决策，同时改善临床实验室工作流程并降低成本。具体目的是：1）开发一种快速单管检测方法，用于同时检测AML和CML/MPD患者的主要突变。2)为检测试剂盒的临床评价制定全面的控制措施。在II期，我们将在诊断实验室环境中评估该检测试剂盒对大量临床样本的诊断价值。我们将开发用于分析检测结果的软件，并协助临床解释结果。最终结果是推出突变特异性分析物特异性试剂（引物，探针），并申请FDA批准作为体外诊断检测。这种用于白血病和MPD突变的检测方法的可用性将大大改善有效治疗白血病和MPD患者所需的临床决策，同时改善临床实验室工作流程并降低成本。该提案的总体目标是开发一种检测突变的方法，该方法对于确定骨髓增生性疾病和白血病的治疗非常重要。该测定的这种液体珠阵列形式将为诊断实验室提供效率和成本效益，并为医生和患者提供用于做出治疗决策的关键信息。