DNA DAMAGE IN SINGLE CELLS--A BIOMARKER OF EXPOSURE

用于对可能暴露于以下物质的人类进行DNA损伤的生物监测： 遗传毒性剂，最佳方法应该能够检测许多 在各种细胞类型的DNA损伤的类别，应提供数据， 单个细胞的水平，并应敏感，快速， 成本效益。 碱性微凝胶电泳分析已被 似乎是为了实现这些目标而开发的。 单细胞凝胶 (SCG)检测，在个别细胞，单链的存在 断裂和碱不稳定位点。 这项技术的其他优点 包括：（1）获得了DNA在细胞间分布的数据 损伤;（ii）需要极少量的细胞（即，几 （iii）其速度、灵敏度及简单性。 的 拟议的研究将进一步表征这一适用性， 技术作为人体细胞DNA损伤的生物标志物 并将评估敏感性差异的方法扩展到 个体间的遗传毒性物质。 当完全特征化时， 这项技术将允许根据血液监测人口 通过针刺手指获得样本。 这一能力将导致 该技术在临床、环境和 职业医学

For the biomonitoring of DNA damage in humans potentially exposed to genotoxic agents, and optimal method should be capable of detecting many classes of DNA damage in a variety of cell types, should provide data at the level of the individual cell, and should be sensitive, rapid and cost effective. An alkaline microgel electrophoresis assay has been developed which appears to fulfill these goals. The Single Cell Gel (SCG) assay detects, in individual cells, the presence of single strand breaks and alkali-labile sites. Other advantages of this technique include: (1) data are obtained on the intercellular distribution of DNA damage; (ii) extremely small numbers of cells are required (i.e., a few thousand cells); and (iii) its speed, sensitivity and simplicity. The proposed research will further characterize the applicability of this technique as a biomarker for DNA damage in the somatic cells of humans and extend the methodology for evaluating differences in sensitivity to genotoxic agents among individuals. When fully characterized, this technique will allow the monitoring of human populations based on blood samples obtained by fingerprick. This capability should result in the widespread application of this technique in clinical, environmental and occupational medicine.