目的 建立高通量評價(jià)藥物遺傳毒性的微型細(xì)菌回復(fù)突變（Ames）試驗(yàn)。方法 采用平皿摻入法進(jìn)行標(biāo)準(zhǔn)Ames試驗(yàn)，采用6孔板摻入法進(jìn)行微型Ames試驗(yàn)，兩試驗(yàn)均設(shè)置陰性對照組和陽性對照組，2組又分別分為代謝活化和非活化2個(gè)亞組，各組培養(yǎng)基中分別加入菌液（鼠傷寒沙門菌組氨酸缺陷型菌株：TA97、TA98、TA100、TA102和TA1535），活化組加大鼠肝微粒體酶（S9）混合液，陽性對照組再加入陽性誘變劑：Dexon（-S9），應(yīng)用于TA97、TA98和TA102；NaN₃（-S9），應(yīng)用于TA100和TA1535；2-AA（+S9），應(yīng)用于TA97、TA98、TA100、TA102和TA1535。凝固后37℃溫箱培養(yǎng)約48 h，計(jì)數(shù)各組回變菌落數(shù)。結(jié)果 在標(biāo)準(zhǔn)Ames試驗(yàn)中，陰性對照組各個(gè)菌株回變菌落數(shù)均在本實(shí)驗(yàn)室自發(fā)回變菌落數(shù)正常值范圍內(nèi)。在兩試驗(yàn)中，陽性對照組所誘導(dǎo)的回變菌落數(shù)均是陰性對照組的2倍以上，標(biāo)準(zhǔn)試驗(yàn)細(xì)菌回變菌落數(shù)約為微型試驗(yàn)的5倍。結(jié)論 在6孔板上進(jìn)行微型細(xì)菌回復(fù)突變試驗(yàn)，大大減少了受試物的用量，提高了篩選速度。本實(shí)驗(yàn)室建立的微型細(xì)菌回復(fù)突變試驗(yàn)背景數(shù)據(jù)，用于遺傳毒理的檢測是高效可靠的。

Objective To establish the method of bacterial reverse-mutation test for high throughput evaluation of drug genotoxicity. Methods The standard Ames test was carried out by the method of plate with incorporation, and the mini-Ames test was performed using 6-well micro plates with incorporation. We established the negative control group and positive control group with metabolic activation or not. The bacteria solution (TA97, TA98, TA100, TA102, and TA1535) was added to the medium respectively. Rat liver microsomal enzyme (S9) mixture was added to the activation group, and the positive control group was added with the positive mutagens:Dexon (-S9) applied to TA97, TA98, and TA102, NaN₃ (-S9) applied to TA100 and TA1535, and 2-AA (+S9) applied to TA97, TA98, TA100, TA102, and TA1535. After incubated, culture plates incubated at 37℃ for about 48 h, counting the number of colonies in each group. Results In the standard Ames test, the number of bacterial colonies in the negative control group was all within the normal range of spontaneous colonies in the laboratory. The number of colonies induced by the positive control group was over two times than that of the negative control group. And the number of colonies in classical test was about five times than that of the mini-Ames. Conclusion The mini-Ames test on 6-well micro plates greatly reduced the dosage of the subject matter and improved the screening speed. The background data of mini-Ames test had been established in our laboratory and this test was highly effective and reliable for the detection of genetic toxicology.

國家科技重大新藥創(chuàng)制項(xiàng)目（2015ZX09501004）；天津市科技計(jì)劃項(xiàng)目（16PTGCCX00090）