Bajpayee Mahima, Kumar Ashutosh, Dhawan Alok
Institute of Life Sciences, School of Science and Technology, Ahmedabad University, Ahmedabad, Gujarat, India.
Methods Mol Biol. 2013;1044:325-45. doi: 10.1007/978-1-62703-529-3_17.
Rapid industrialization and pursuance of a better life have led to an increase in the amount of chemicals in the environment, which are deleterious to human health. Pesticides, automobile exhausts, and new chemical entities all add to air pollution and have an adverse effect on all living organisms including humans. Sensitive test systems are thus required for accurate hazard identification and risk assessment. The Comet assay has been used widely as a simple, rapid, and sensitive tool for assessment of DNA damage in single cells from both in vitro and in vivo sources as well as in humans. Already, the in vivo comet assay has gained importance as the preferred test for assessing DNA damage in animals for some international regulatory guidelines. The advantages of the in vivo comet assay are its ability to detect DNA damage in any tissue, despite having non-proliferating cells, and its sensitivity to detect genotoxicity. The recommendations from the international workshops held for the comet assay have resulted in establishment of guidelines. The in vitro comet assay conducted in cultured cells and cell lines can be used for screening large number of compounds and at very low concentrations. The in vitro assay has also been automated to provide a high-throughput screening method for new chemical entities, as well as environmental samples. This chapter details the in vitro comet assay using the 96-well plate and in vivo comet assay in multiple organs of the mouse.
快速工业化以及对美好生活的追求导致环境中化学物质的数量增加，这些化学物质对人类健康有害。农药、汽车尾气和新型化学物质都会加剧空气污染，并对包括人类在内的所有生物产生不利影响。因此，需要灵敏的检测系统来进行准确的危害识别和风险评估。彗星试验已被广泛用作一种简单、快速且灵敏的工具，用于评估来自体外、体内以及人类的单细胞中的DNA损伤。目前，体内彗星试验作为评估动物DNA损伤的首选试验，在一些国际监管指南中已变得至关重要。体内彗星试验的优点在于，尽管存在非增殖细胞，它仍能够检测任何组织中的DNA损伤，并且对检测遗传毒性具有敏感性。针对彗星试验举办的国际研讨会所提出的建议已促成了相关指南的制定。在培养细胞和细胞系中进行的体外彗星试验可用于筛选大量化合物，且浓度极低。体外试验也已实现自动化，为新型化学物质以及环境样品提供了一种高通量筛选方法。本章详细介绍了使用96孔板进行的体外彗星试验以及在小鼠多个器官中进行的体内彗星试验。
