Molecular Diagnostics Core Laboratory

分子诊断核心实验室

基本信息

批准号：
7733475
负责人：
Mark Raffeld
金额：
$ 142.26万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
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/7733475
关键词：
Accounting American Area Biological Assay Blood specimen Body Fluids Capillary Electrophoresis Categories Cerebrospinal Fluid Childhood Classification Clinical Clinical Trials Clonality Complex DNA Data Diagnostic EWS-FLI1 fusion protein Eligibility Determination Epidermal Growth Factor Receptor Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Freezing Funding Genes Human Herpesvirus 4 Human Herpesvirus 8 Human T-lymphotropic virus 1 Individual Laboratories Lymphoma Lymphoproliferative Disorders MGMT gene Malignant Neoplasms Metabolism Methylation Microdissection Molecular Molecular Abnormality Molecular Diagnostic Testing Molecular Target Mutation Mutation Analysis National Institute of Allergy and Infectious Disease Nucleic Acids Numbers Paraffin Paraffin Embedding Pathologist Pathology Patients Polymerase Chain Reaction Process Proteins Publications Purpose RNA Research Personnel Research Project Grants Resources Reverse Transcriptase Polymerase Chain Reaction Sampling Satellite Viruses Source Specimen Structure Study Section T-Lymphocyte Testing Time Tissues Translational Research United States National Institutes of Health Viral Load result clinically relevant college molecular oncology prognostic promoter response sarcoma tissue fixing

项目摘要

The molecular diagnostics laboratory processed approximately 900 unique clinical samples in 2007, accounting for over 3000 tests. Approximately 300 additional 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. 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 3 Perkin-Elmer 9700 PCR platforms, one ABI 7900 real-time PCR machine, and one ABI 3130 capillary electrophoresis unit. As part of the recent funding initiative to expand molecular diagnostics testing in the LP, the laboratory just purchased a Pyromark 2400 pyrosequencer which will be employed initially for K-RAS mutation analysis, and for MGMT promoter methylation analysis. 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 and data to support their publications are: Dr. Elaine Jaffe (NCI) Dr. Maria Tsokos (NCI) Dr. Andrea Abati (NCI) Dr. Robert Kreitman (NCI) Dr. Wyndham Wilson (NCI) Dr. John Janik (NCI) Dr. John Morris (NCI) Dr. Steven Holland (NIAID) Dr, Richard Little (NCI) Dr. Dean Metcalfe (NIAID) Dr. Amy Klion Dr. John Morris

分子诊断实验室在2007年处理了大约900个独特的临床样本，占3000多次测试。进行了大约300次其他测试，以支持NCI/NIH PIS的各种翻译研究项目。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。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3个Perkin-Elmer 9700 PCR平台，一台ABI 7900实时PCR机和一台ABI 3130毛细管电泳单元。作为扩大LP分子诊断测试的最近资金计划的一部分，该实验室刚刚购买了Pyromark 2400 Pyrosequerquer，该pyrosequerquer最初将用于K-RAS突变分析和MGMT启动子甲基化分析。分子诊断实验室支持NCI和NIH研究人员的转化研究。在NCI和NIH任职期限的研究人员和临床医生中，利用实验室的资源和数据来支持其出版物：Elaine Jaffe博士（NCI）Maria Tsokos博士（NCI）博士（NCI）Andrea Abati（NCI）Andrea Abati（NCI）Robert Kreitman博士（NCI）荷兰（Niaid）Dr，Richard Little（NCI）Dean Metcalfe博士（Niaid）Amy Klion博士John Morris博士