Development of a standard high throughput comet assay

标准高通量彗星测定的开发

基本信息

批准号：
7287399
负责人：
Jay George
金额：
$ 44.86万
依托单位：
TREVIGEN, INC.
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-01-01 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7287399
关键词：
Achievement Acute Address Adoption Affect Age Alkaline Single-Cell Gel Electrophoresis Assay Anodes Antigens Antineoplastic Agents Appendix Archives Area Attention Automation Benzopyrenes Biological Biological Assay Biological Monitoring Biological Preservation Buffers Carcinogens Cell Count Cell Cycle Cell Cycle Arrest Cell Death Cell Line Cells Chemicals Chronic Cleaved cell Collecting Cell Comet Assay Communities Computer software Condition Consensus Contracts Controlled Environment Critiques Cytolysis DNA DNA Adducts DNA Damage DNA Repair DNA analysis DNA glycosylase Data Data Analyses Deposition Detection Detergents Development Development Plans Device or Instrument Development Educational workshop Electrophoresis Engineering Ensure Environmental Monitoring Epidemiology Epoxy Compounds Equipment Exposure to Feedback Fluorescence Fluorescent Dyes Fluorescent in Situ Hybridization Funding Future Gel Genes Genetic Germany Glass Glycol Goals Grant Guidelines Hand Height Imagery Immobilized Cells Immunohistochemistry In Situ In Situ Hybridization Individual International Laboratories Lead Learning Letters Manuals Manufacturer Name Market Research Marketing Metabolic Methods Microscope Modification Monitor Movement Mutagens Mutation Mycoplasma Nature Normal Cell Numbers Nutritional Olives - dietary Operative Surgical Procedures Patients Pattern Performance Persons Pharmaceutical Preparations Pharmacologic Substance Phase Phase I Clinical Trials Positioning Attribute Preclinical Drug Evaluation Preparation Price Procedures Process Progress Reports Protocols documentation Publications Published Comment Publishing Pump Purpose Radiation Range Rate Reagent Recommendation Records Reporting Reproducibility Research Research Personnel Resistance Retirement Risk Assessment Running Sales Sampling Scientist Screening procedure Sepharose Signal Transduction Site Slide Small Business Funding Mechanisms Small Business Innovation Research Grant Sodium Chloride Source Specific qualifier value Staining method Stains Standardization Standards of Weights and Measures Structure Suggestion System Tail Techniques Technology Temperature Testing Text Time Tissues Toxicology Treatment Effectiveness Treatment outcome United States Food and Drug Administration Validation Variant Virus Work adduct anticancer research base cancer cell cancer risk cancer therapy carcinogenesis cell preparation cell type chemotherapeutic agent commercialization cost cost effective crosslink day design drug development drug discovery electric field endonuclease gel electrophoresis genotoxicity high throughput analysis improved instrument instrumentation interest melting migration neoplastic cell pre-clinical programs prototype reagent standardization repaired research study response sensor therapy resistant tumorigenesis voltage

项目摘要

DESCRIPTION (provided by applicant): The single cell gel electrophoresis (SCGE) or comet assay is now recognized worldwide as a simple and inexpensive method for detection of different types of DNA damage. Adoption of the comet assay for genotoxic studies has been hampered to date by the lack of a standardized platform and procedures for the assay, as well as the labor-intensive nature of the assay for multiple-sample analysis. Another problem associated with the assay is that to avoid introduction of artificial DNA damage to cells, the assay needs to be run of freshly isolated cells or tissues. This make it difficult to direct compare cells collected at various times, or to collect samples in the field where the assay cannot immediately performed. Trevigen in the Phase I application developed a high throughput comet slides for standardization of the comet slide preparation for the assay. These slides are available at present in formats for single assay analysis (2 well format) and also for high throughput analysis (20 and 96 well format). Trevigen also demonstrated in preliminary studies that it is possible to preserve cells without DNA damage for analysis later or to use as standards in subsequent assays. The goals of the Phase II application are: 1. To standardize the electrophoretic platform and procedures for the comet assay by developing a controlled environment for electrophoresis of the damaged DNA in the cells, which will eliminate the problems, associated with reproducibility of the comet assay in the day to day operation. 2. To develop comet assay standards based on cells, which were treated with various concentration of DNA damaging agents, and long term preservation of these standards or other biological materials, to be used as reference material in the comet assay. The accomplishment of these aims will results in a standardized comet assay procedure, which can be used in any laboratory and can be developed as a high throughput automatic system. The preservation of cells without DNA damage will allow development of standards for the comet assay and will allow that biological material could be archived for future studies or to be shared between laboratories. It would also allow field collected samples to be safely transported for analysis later. The single cell gel electrophoresis (SCGE) or comet assay is a simple and inexpensive method for detection of different types of DNA damage. The long-term objective of this project is to develop standardized platform and procedures, including DNA standards for this assay.

描述（由申请人提供）：单细胞凝胶电泳（SCGE）或彗星测定现在在全球范围内被视为一种简单且廉价的方法，用于检测不同类型的DNA损伤。迄今为止，由于缺乏标准化平台和测定程序的程序以及用于多样本分析的劳动密集型性质，因此对遗传毒性研究的采用受到了阻碍。与该测定有关的另一个问题是，要避免引起人工DNA对细胞的损害，该测定必须由新鲜分离的细胞或组织进行。这使得很难指导在不同时间收集的细胞，或者在无法立即执行测定的现场收集样品。 I阶段应用程序中的Trevigen开发了高通量彗星幻灯片，用于标准化测定的彗星幻灯片准备。这些幻灯片目前以单个测定分析（2孔格式）以及高吞吐量分析（20和96井格式）提供。 Trevigen在初步研究中还证明，可以保留无DNA损伤的细胞以稍后分析或在随后的测定中用作标准。 II期应用的目标是：1。通过开发细胞中受损DNA的电泳的受控环境，标准化电泳平台和彗星测定过程，这将消除与日常运行中彗星测定的可重复性有关的问题。 2。开发基于细胞的彗星测定标准，并用各种浓度的DNA损害剂处理，并长期保留这些标准标准或其他生物材料，以作为彗星测定中的参考材料。这些目标的实现将导致标准化的彗星测定程序，该过程可用于任何实验室，可以作为高通量自动系统开发。保存没有DNA损伤的细胞将允许开发彗星测定的标准，并允许可以将生物材料存档以进行未来的研究或在实验室之间共享。这也将允许采集的现场样品安全运输以供以后进行分析。单细胞凝胶电泳（SCGE）或彗星测定是一种简单且廉价的方法，用于检测不同类型的DNA损伤。该项目的长期目标是开发标准化的平台和程序，包括该测定法的DNA标准。