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彗星试验在化合物遗传毒性评价中的应用日益广泛,ICH S2(R1)已将肝脏彗星试验列为第2个组织/终点的体内试验;经济合作与发展组织(Organization for Economic Co-operation and Development,OECD)体内哺乳动物碱性彗星试验的指导原则(TG489)也已颁布,但彗星试验的具体操作程序各异,需要严格控制的影响因素较多,且因组织差异性,国际上无统一的标准试验方案。毒理工作者们对该试验方法的不同影响因素进行研究,以望能够形成标准的试验方案,在不影响灵敏度和特异性的前提下,提高彗星试验的可操作性,推动彗星试验在评价化合物遗传毒性方面更广泛的应用。本文主要对彗星试验的取样、制片、解旋和电泳等过程中关键的影响因素进行探讨总结,为开展和验证彗星试验方法的实验室提供参考。
The Comet Assay has become more and more widely used in the evaluation of the genotoxicity of compounds. ICH S2 (R1) has listed the liver comet assay as the second tissue / endpoint in vivo test. The Organization for Economic Co-operation and Development OECD Guidelines for In vivo Mammalian Alkaline Comet Assays (TG489) have also been published, but the specific procedures for controlling comet assay vary and many factors that need to be tightly controlled are high. There is no internationally agreed standard due to organizational differences Test plan. Toxicological workers study the different influencing factors of this test method in the hope of forming a standard test protocol and improving the operability of the comet assay without affecting the sensitivity and specificity, Genotoxicity is more widely used. In this paper, the key influencing factors in the process of sampling, preparation, spinning and electrophoresis of comet assay are discussed and summarized, which will provide reference for the laboratory to carry out and verify comet assay.