Molecular Diagnostics Core Laboratory

分子诊断核心实验室

• 批准号: 7970028
• 负责人: Mark Raffeld
• 金额: $ 50.41万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7970028
• 关键词: Accounting; American; Area; BRAF gene; Biological Assay; Biological Markers; Blood specimen; Body Fluids; Cancer Diagnostics; Capillary Electrophoresis; Categories; Cerebrospinal Fluid; Childhood; Classification; Clinical; Clinical Trials; Clonality; Colon; Complex; DNA; Development; Diagnostic; EWS-FLI1 fusion protein; Eligibility Determination; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Freezing; Genes; Human Herpesvirus 8; Human T-lymphotropic virus 1; Individual; KRAS2 gene; Laboratories; Lymphoma; Lymphoproliferative Disorders; MGMT gene; Malignant Neoplasms; Malignant neoplasm of brain; Malignant neoplasm of lung; Metabolism; Methylation; Microdissection; Molecular; Molecular Abnormality; Molecular Diagnostic Testing; Molecular Target; Mutation; Mutation Analysis; National Heart, Lung, and Blood Institute; National Human Genome Research Institute; National Institute of Allergy and Infectious Disease; Nucleic Acids; PIK3CA gene; Paraffin; Paraffin Embedding; Pathologist; Pathology; Patients; Process; Proteins; Proto-Oncogene Proteins c-akt; RNA; Research; Research Personnel; Research Project Grants; Resources; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Satellite Viruses; Source; Specimen; Structure; Study Section; T-Lymphocyte; Technology; Testing; Time; Tissues; Translational Research; United States National Institutes of Health; Viral Load result; austin; clinically relevant; college; epidermal growth factor receptor VIII; instrument; molecular oncology; next generation; prognostic; response; sarcoma; tissue fixing; tumor

The molecular diagnostics laboratory processed approximately 1011 unique clinical samples in 2008 on NCI/NIH patients, accounting for over 3500 tests. Another 300+ tests were performed to support a variety of translational research projects, for NCI/NIH PIs. The majority of the laboratory's tests are conventional PCR assays, including both DNA and RT-PCR assays. The assays performed by the molecular diagnostics laboratory include assays for B and T-cell clonality, assays for lymphoma related translocations, assays for cancer associated viruses (EBV, HTLV, HHV-8), and RT-PCR assays for translocations associated with pediatric sarcomas. Quantitative PCR is performed for HTLV 1/2 to follow viral load for the Metabolism Branch's ATL clinical trials. KRAS and EGFR mutation analysis is performed to assess eligibility and response to EGFR-targeted therapies. Currently we are developing assays to assess BRAF, AKT and PIK3CA mutation, EGFRvIII presence, and MGMT methylation in a variety of cancers. The laboratory's developmental research effort is focused on the application of next generation sequencing technologies for clinical use, and the study of circulating DNA biomarkers for cancer diagnostics. Assays are performed on materials derived from a variety of sources. Paraffin fixed tissue specimens account for about 60% of the samples and blood, 30%. The remaining 10% of samples are derived from fresh or frozen tissue, cerebrospinal fluid, and other body fluids. A high percentage of the paraffin section studies are complex and labor intensive, requiring macro or microdissection of specific components of lesional tissues, and are performed under the guidance of a fellow or staff pathologist. The molecular laboratory is particularly expert in extracting nucleic acids from small fragments of archival paraffin-embedded tissues, and our expertise in this area has been widely sought out in a number of laboratories within and outside of NIH. The laboratory maintains 4 Perkin-Elmer 9700 PCR platforms, one ABI 7900 real-time PCR machine, one ABI 3130xl capillary electrophoresis unit, and one Pyromark 24 pyrosequencing instrument. The molecular diagnostics laboratory supports translational research of both NCI and NIH researchers. Among the NCI and NIH tenured investigators and clinicians that have utilized the laboratory's resources are: Dr. Austin Barrett (NHLBI) Dr. Susan Bates (NCI) Dr. John Bennett (NIAID) Dr. Michael Bishop (NCI) Dr. Daniel Fowler (NCI) Dr. William Gahl (NHGRI) Dr. Giuseppe Giaccone (NCI) Dr. Steven Holland (NIAID) Dr. Elaine Jaffe (NCI) Dr. John Janik (NCI) Dr. Amy Klion (NHLBI) Dr. Robert Kreitman (NCI) Dr. Shivaani Kummar (NCI) Dr, Richard Little (NCI) Dr. Gerald Marti (FDA) Dr. Dean Metcalfe (NIAID) Dr. John Morris (NCI) Dr. Kenneth Olivier (NIAID) Dr. Stefania Pittaluga (NCI) Dr. Koneti Rao (NIAID) Dr. Phillip Scheinberg (NHLBI) Dr. Maria Tsokos (NCI) Dr. Maria Turner (NCI) Dr. Thomas Uldrick (NCI) Dr. Gulbu Uzel (NIAID) Dr. Wyndham Wilson (NCI) Dr. Neil Young (NHLBI)

分子诊断实验室在2008年对NCI/NIH患者进行了大约1011个独特的临床样本处理，占3500多次测试。对NCI/NIH PIS进行了另外300多次测试，以支持各种翻译研究项目。实验室的大多数测试是常规的PCR分析，包括DNA和RT-PCR分析。分子诊断实验室进行的测定包括B和T细胞克隆性的测定，相关淋巴瘤相关易位的测定，癌症相关病毒的测定法（EBV，HTLV，HHV-8）和与儿科肉瘤相关的易位的RT-PCR测定法。对HTLV 1/2进行定量PCR，以遵循代谢分支的ATL临床试验的病毒载量。进行KRAS和EGFR突变分析以评估对EGFR靶向疗法的资格和反应。目前，我们正在开发用于评估BRAF，AKT和PIK3CA突变，EGFRVIII的存在和MGMT甲基化的测定法。实验室的发展研究工作重点是将下一代测序技术用于临床使用，以及研究用于癌症诊断的循环DNA生物标志物。对来自各种来源的材料进行测定。石蜡固定组织标本约占样品和血液的60％，为30％。 其余10％的样品来自新鲜或冷冻的组织，脑脊液和其他体液。石蜡部分的很高比例是复杂且劳动密集型的，需要对病变组织的特定组成部分进行宏观或显微解剖，并且在同胞或职员病理学家的指导下进行。该分子实验室特别是从档案型石蜡包裹的组织的小碎片中提取核酸的专家，我们在该领域的专业知识已在NIH内外的许多实验室中广泛寻求。该实验室维护4个Perkin-Elmer 9700 PCR平台，一台ABI 7900实时PCR机器，一台ABI 3130xl毛细管电泳单元和一个Pyromark 24 PyroMark 24 Pyrosequencen仪器。分子诊断实验室支持NCI和NIH研究人员的转化研究。在利用实验室资源的NCI和NIH终身调查人员和临床医生中，Austin Barrett博士（NHLBI）Susan Bates博士（NCI）John Bennett博士（NIAID）博士Michael Bishop（NCI）（NCI）博士（NCI） （Niaid）Elaine Jaffe博士（NCI）John John Jaffe（NCI）博士Amy Klion博士（NHLBI）Robert Kreitman博士（NCI）Shivaani Kummar博士（NCI）博士（NCI），Richard Little（NCI） （Niaid）Stefania Pittaluga博士（NCI）Koneti Rao（Niaid）博士Phillip Scheinberg博士（NHLBI）玛丽亚·托科斯（NHLBI）博士（NCI）玛丽亚·特纳（NCI）玛丽亚·特纳（NCI）博士（NCI）托马斯·托马斯·乌德里克（NCI）博士（NCI） （NHLBI）