Koppen Gudrun, Azqueta Amaya, Pourrut Bertrand, Brunborg Gunnar, Collins Andrew R, Langie Sabine A S
Environmental Risk and Health unit, Flemish Institute of Technological Research (VITO), Boeretang 200, 2400 Mol, Belgium.
Department of Pharmacology and Toxicology, University of Navarra, and IdiSNA, Navarra Institute for Health Research, C/Irunlarrea 1, 31009 Pamplona, Spain.
Mutagenesis. 2017 May 1;32(3):397-408. doi: 10.1093/mutage/gex002.
The International Comet Assay Workshops are a series of scientific conferences dealing with practical and theoretical aspects of the Comet Assay (single-cell gel electrophoresis)-a simple method for detecting DNA strand breaks. The first paper describing such an assay was published over 30 years ago in 1984 by Swedish researchers O. Ostling and K. J. Johanson. Appropriately, the theme for the 2015 meeting was looking to the future: 'The Next 3 Decades of the Comet Assay'. The programme included 25 oral and 43 poster presentations depicting the latest advances in technical developments as well as applications of the comet assay in genotoxicity testing (in vitro and in vivo) and biomonitoring of both humans and the environment. Open discussion sessions based on questions from the participants allowed exchange of practical details on current comet assay protocols. This report summarises technical issues of high importance which were discussed during the sessions. We provide information on ways to improve the assay performance, by testing for cytotoxicity, by using reference samples to reduce or allow for inter-experimental variation, and by standardising quantification of the damage, including replicates and scoring enough comets to ensure statistical validity. After 30 years of experimentation with the comet assay, we are in a position to control the important experimental parameters and make the comet assay a truly reliable method with a wealth of possible applications.
国际彗星试验研讨会是一系列科学会议，涉及彗星试验（单细胞凝胶电泳）的实践和理论方面，这是一种检测DNA链断裂的简单方法。第一篇描述这种试验的论文是30多年前由瑞典研究人员O. 奥斯特林和K. J. 约翰森于1984年发表的。恰如其分地，2015年会议的主题是展望未来：“彗星试验的下一个30年”。会议议程包括25个口头报告和43个海报展示，展示了技术发展的最新进展以及彗星试验在遗传毒性测试（体外和体内）以及人类和环境生物监测中的应用。基于参会者提出的问题进行的公开讨论环节，促进了关于当前彗星试验方案实际细节的交流。本报告总结了会议期间讨论的高度重要的技术问题。我们提供了有关提高试验性能的方法的信息，包括通过检测细胞毒性、使用参考样品以减少或考虑实验间差异，以及通过对损伤进行标准化定量，包括重复实验和对足够数量的彗星进行评分以确保统计有效性。经过30年对彗星试验的实验，我们能够控制重要的实验参数，并使彗星试验成为一种具有大量可能应用的真正可靠的方法。
