体内彗星试验方法学研究进展  

Progress in the methodological study of in vivo Comet assay

作　　者：欧红梅[1] 周长慧[1] 涂宏刚[1] 常艳[1] 

机构地区：[1]中国医药工业研究总院、上海医药工业研究院、国家上海新药安全评价研究中心,上海201203

出　　处：《中国新药杂志》2014年第17期1990-1993,共4页Chinese Journal of New Drugs

基　　金：国家“重大新药创制”科技重大专项(2012ZX09302002)

摘　　要：体内彗星试验是一种简单的DNA损伤检测方法,可检测单链断裂、双链断裂、碱不稳定位点、DNA-DNA和DNA-蛋白质交联等。2011年11月,修订完成的ICH S2(R1)指导原则将体内彗星试验列入遗传毒性标准组合体内首选的第2个检测终点,故规范试验操作方法具有重要意义。该文主要依据JaCVAM联合验证试验,从给药方案设计、取样、样品处理、电泳,DNA损伤检测指标及可接受标准6个方面阐述目前公认的体内彗星试验方法,为该试验用于药物新药安全性评价研究等领域提供方法学参考。此外,本文就体内彗星试验的主要影响因素进行了探讨,希望进一步提高该试验准确性和可重复性。The in vivo Comet assay is a simple method for measuring DNA damage, such as single strand breaks, double strand breaks, alkali labile sites, DNA-DNA and DNA-protein crosslinks. It is a useful method for screening and evaluation of the genetic toxicity of new drugs and prediction of carcinogenic compounds. With in- creasing application in detecting genotoxicity damage, and being listed in ICH $2 （ R1 ） guidelines in genetic toxici- ty standard combinatorial test, the standardization of the assay operation has important significance. This paper is mainly based on JaCVAM international validation study. The treatment schedule, tissue collection, preparation of specimens, unwinding and electrophoresis, endpoints of DNA damage and the acceptance criteria were described to provide methodology reference for safety evaluation and research of new drugs. What＇s more, we discussed the main influencing factors of in vivo comet assay to further improve the assay accuracy and repeatability.

关 键 词：体内彗星试验 DNA损伤 方法学 影响因素 

分 类 号：R99[医药卫生—毒理学]