Automation of the vitro Micronucleus Assay

体外微核测定的自动化

• 批准号: 6792136
• 负责人: STEPHEN D DERTINGER
• 金额: $ 15.42万
• 依托单位: LITRON LABORATORIES, LTD.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6792136
• 关键词: DNA damage; cell line; chromosomes; flow cytometry; fluorescent dye /probe; method development; micronucleus; microscopy; terminal nick end labeling; tissue /cell culture

DESCRIPTION (provided by applicant): The induction of DNA damage and the resulting sequelae of mutations and chromosomal rearrangements are primary mechanisms by which cancers arise. DNA damage has also been implicated in arteriosclerosis and certain birth defects. Thus, there is an important need for sensitive methods for identifying compounds that mutate or otherwise damage DNA. The very high cost of long-term animal studies dictates that short- and medium-term mutagenicity assays must be relied on to eliminate potentially hazardous chemicals from development, and to prioritize chemicals for further study. Furthermore, as new chemicals and therapeutic drugs are being developed ever more rapidly, the need for short-term tests with high throughput capabilities increases. The aim of this Phase II research effort is to refine, standardize, and validate an automated system for scoring chromosome damage in cultured mammalian cells. Specifically, a flow cytometric method for enumerating genotoxicant-induced micronuclei will be developed and vigorously tested. This system will be extremely valuable as an early screening test for identifying genotoxicants, and is expected to prove particularly valuable to the Chemical Manufacturing and Pharmaceutical Industries which are required to assess compounds' DNA-damaging potential. The automated scoring systems' commercial potential will be realized through: 1) fee-for-service contract testing, and 2) bundling dyes and other reagents required to perform these analyses into commercially available kits.

描述（由申请人提供）： DNA损伤的诱导以及由此产生的突变和染色体重排的后遗症是癌症发生的主要机制。DNA损伤也与动脉硬化和某些出生缺陷有关。因此，非常需要用于鉴定突变或以其他方式损害DNA的化合物的灵敏方法。长期动物研究的成本非常高，因此必须依靠短期和中期致突变性试验来消除开发中的潜在危险化学品，并优先考虑进一步研究的化学品。此外，随着新的化学品和治疗药物的开发越来越快，对具有高通量能力的短期测试的需求也在增加。这项第二阶段研究工作的目的是完善，标准化和验证一个自动化系统，用于对培养的哺乳动物细胞中的染色体损伤进行评分。具体而言，将开发用于计数遗传毒性诱导微核的流式细胞术方法，并进行严格检测。该系统将是非常有价值的早期筛选试验，用于识别遗传毒物，并预计将证明特别有价值的化学制造和制药工业，需要评估化合物的DNA损伤的潜力。自动评分系统的商业潜力将通过以下方式实现：1）按服务付费的合同测试，以及2）将进行这些分析所需的染料和其他试剂捆绑到市售试剂盒中。