Bajpayee Mahima, Kumar Ashutosh, Dhawan Alok
, New Delhi, India.
Biological and Life Sciences, School of Arts and Sciences, Ahmedabad University, Ahmedabad, Gujarat, India.
Methods Mol Biol. 2019;2031:237-257. doi: 10.1007/978-1-4939-9646-9_12.
Anthropogenic activities, indiscriminate and rapid industrialization as well as pursuance of a better life has led to an increase in the concentration of chemicals, like pesticides, automobile exhausts, and new chemical entities, in the environment, which have an adverse effect on all living organisms including humans. Sensitive and robust 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 cell from both in vitro and in vivo sources as well as in humans. The advantages of the in vivo Comet assay are its ability to detect DNA damage in any tissues, 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, and the OECD has adopted a guideline for the in vivo Comet assay as a test for assessing DNA damage in animals. The in vitro Comet assay conducted in cultured cells 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 in 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.
人为活动、无节制且迅速的工业化以及对美好生活的追求，导致环境中化学物质（如农药、汽车尾气和新型化学物质）的浓度增加，这些化学物质对包括人类在内的所有生物都有不利影响。因此，需要灵敏且可靠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，用于评估来自体外和体内来源以及人类的单细胞中的DNA损伤。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。为彗星试验举办的国际研讨会所提出的建议促成了相关指南的制定，经济合作与发展组织（OECD）已采用一项体内彗星试验指南，作为评估动物DNA损伤的一项检测方法。在培养细胞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。
