目的 使用鼠傷寒沙門菌和大腸埃希菌及陽性劑開展基于6孔板、24孔板和標(biāo)準(zhǔn)10 cm平皿的Ames試驗(yàn)，為確立標(biāo)準(zhǔn)化微孔板Ames試驗(yàn)奠定研究基礎(chǔ)。方法 6種不同鼠傷寒沙門菌（TA97、TA98、TA100、TA102、TA1535、TA1537）和1種大腸埃希菌（WP2 uvrA）經(jīng)鑒定及擴(kuò)增后，分別使用標(biāo)準(zhǔn)平皿、6孔板和24孔板在有無S9代謝活化條件下使用不同濃度的陽性劑開展平板摻入法Ames試驗(yàn)。所有樣本置37℃培養(yǎng)約48 h后進(jìn)行菌落計(jì)數(shù)。結(jié)果 所有試驗(yàn)條件下均出現(xiàn)濃度相關(guān)性的回復(fù)菌落形成率增加，且最高濃度條件下的菌落數(shù)高于對(duì)照組3倍。然而微孔板中可出現(xiàn)陰性背景菌落數(shù)過少的情況，陽性劑的用量也不能照搬標(biāo)準(zhǔn)平皿中的濃度設(shè)置。結(jié)論 本研究所用條件可成功開展基于6孔板和24孔板的Ames試驗(yàn)，其試驗(yàn)條件和背景數(shù)據(jù)需要經(jīng)過摸索與積累。

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.

國(guó)家"重大新藥創(chuàng)制"科技重大專項(xiàng)（2018ZX09201017）；國(guó)家自然科學(xué)基金（81503347）