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: 6949034
• 负责人: George L. DeGeorge
• 金额: $ 60.12万
• 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 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6949034
• 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）：工业上越来越需要一种廉价的筛选试验，该试验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。程序外DNA合成（UDS）试验是一种经验证的鉴定和表征遗传毒性化学品的方法。然而，标准测定的几个技术方面限制了其广泛使用。我们通过将流式细胞术作为检测方法，对I期研究中的标准UDS测定进行了修改，并证明了我们优化版本的UDS（称为FLUDS）的可行性。FLUDS检测允许更高的通量和更短的研究时间（标准UDS为几天而不是几周）。这种新的检测方法可以区分先天性和生物转化的遗传毒物，并完全定量。它可靠地预测了乙基亚硝基脲，阿非迪霉素，2-乙酰氨基芴，芴诱导的UDS。我们打算通过检测一组扩展的遗传毒性和非遗传毒性化学品来预验证FLUDS检测试剂盒，为ICCVAM指导原则下的多实验室验证做准备。预先验证的FLUDS测定将对制药、化学、化妆品和消费品行业的客户具有相当大的商业价值。