目的 使用鼠伤寒沙门菌和大肠埃希菌及阳性剂开展基于6孔板、24孔板和标准10 cm平皿的Ames试验，为确立标准化微孔板Ames试验奠定研究基础。方法 6种不同鼠伤寒沙门菌（TA97、TA98、TA100、TA102、TA1535、TA1537）和1种大肠埃希菌（WP2 uvrA）经鉴定及扩增后，分别使用标准平皿、6孔板和24孔板在有无S9代谢活化条件下使用不同浓度的阳性剂开展平板掺入法Ames试验。所有样本置37℃培养约48 h后进行菌落计数。结果 所有试验条件下均出现浓度相关性的回复菌落形成率增加，且最高浓度条件下的菌落数高于对照组3倍。然而微孔板中可出现阴性背景菌落数过少的情况，阳性剂的用量也不能照搬标准平皿中的浓度设置。结论 本研究所用条件可成功开展基于6孔板和24孔板的Ames试验，其试验条件和背景数据需要经过摸索与积累。

Objectives To compare the Ames tests based on 6-well/24-well plates and standard 10cm plates using Salmonella typhimurium/Escherichia coli and postive agents, and lay a research foundation for the standardized microplate based Ames tests. Methods 6 Salmonella typhimurium (TA97, TA98, TA100, TA102, TA1535, TA1537) and a Escherichia coli (WP2 uvrA) were idenfied and amplified, and subsequently used to perform standard plate, 6-well plate and 24-well plate based Ames tests with a range of different concentrations of positive agents in the presence or absence of S9 metabolic activation. The colonies were counted 48 hours after cultured at 37℃. Results The concentration relevant colony formation rates increasing were observed in all test conditions Under the highest concentrations, the colony counts were 3 times higher than the solvent control. However, too few backgournd colony counts might be found in the solvent controls of microplates, and the amount of positive agents might not be set in accord with the concentrations used in standard plate test. Conclusions 6-well and 24-well plate based Ames tests were performed successfully following the current protocol, whereas the experimental conditions and background data shall be further explored and accumulated.

国家"重大新药创制"科技重大专项（2018ZX09201017）；国家自然科学基金（81503347）