Automation of the vitro Micronucleus Assay

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，重要的是需要灵敏的方法来识别突变或以其他方式损害DNA的化合物。长期动物研究的非常高的成本决定了必须依靠短期和中期的诱变性分析来从开发中消除潜在的危险化学品，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增加了。这项第二阶段研究工作的目的是改进、标准化和验证一种用于对培养的哺乳动物细胞中的染色体损伤进行评分的自动化系统。具体地说，将开发一种流式细胞术方法来计数遗传毒物诱导的微核，并对其进行有力的测试。该系统作为识别基因毒物的早期筛选测试将非常有价值，预计将被证明对化学制造和制药行业特别有价值，这些行业需要评估化合物的DNA损伤潜力。自动评分系统的商业潜力将通过以下方式实现：1)按服务收费的合同测试，以及2)将执行这些分析所需的染料和其他试剂捆绑到商业可用试剂盒中。

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.