Flow cytometry-based Unscheduled DNA Synthesis (FLUDS)

基于流式细胞术的非计划 DNA 合成 (FLUDS)

• 批准号: 6790402
• 负责人: George L. DeGeorge
• 金额: $ 74.77万
• 依托单位: MB RESEARCH LABORATORIES, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-15 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6790402
• 关键词: DNA damage; DNA replication; cell line; chick embryo; flow cytometry; high throughput technology; mutagen testing; nucleic acid biosynthesis; technology /technique development; toxicant screening

DESCRIPTION (provided by applicant): There is a growing need in industry for an inexpensive screening assay which can reliably determine the genotoxic potential of the large number of new chemical compounds synthesized each year. The Unscheduled DNA Synthesis (UDS) Assay is a proven method to identify and characterize genotoxic chemicals. However, several technical aspects of the standard assay limit its widespread use. We have made modifications to the standard UDS assay in Phase I studies through incorporation of flow cytometry as a detection method, and have proven feasibility of our optimized version of UDS, termed FLUDS. The FLUDS assay allows higher throughput and much shorter study times (a few days versus weeks for the standard UDS). This new assay can discriminate between innate and biotransformed genotoxicants, and is fully quantitative. It has reliably predicted the UDS induced by ethylnitrosourea, aphidicolin, 2- acetylaminofluorene, and fluorene. We intend to pre-validate the FLUDS assay by testing an expanded set of genotoxic and non-genotoxic chemicals in preparation for a multi-laboratory validation under ICCVAM guidelines. The pre-validated FLUDS assay would be of considerable commercial value to customers in the pharmaceutical, chemical, cosmetic and consumer products industries.

描述（由申请人提供）：行业对廉价筛选测定法的需求越来越大，可以可靠地确定每年合成的大量新化合物的遗传毒性潜力。外规定的DNA合成（UDS）测定是一种鉴定和表征遗传毒性化学物质的验证方法。但是，标准测定法的几个技术方面限制了其广泛使用。我们通过将流式细胞仪作为检测方法纳入I阶段研究中的标准UDS分析进行了修改，并证明了我们优化的UDS的可行性，称为Fluds。 Fluds测定法可以更高的吞吐量和更短的学习时间（与标准UDS相比几天）。这种新测定可以区分先天和生物转化的遗传毒性，并且是完全定量的。它可靠地预测了乙基硝基乙酸，蚜虫，2-乙酰基氨基氟烯烯和氟的UD。我们打算通过测试一组扩展的遗传毒性和非基因毒性化学物质来预验证FLUDS测定法，以准备在ICCVAM指南下进行多种实验室验证。预先验证的Fluds分析对于药品，化学，化妆品和消费产品行业的客户来说将具有可观的商业价值。