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转基因产品成分检测是转基因生物安全管理和标识的重要支撑,而转基因生物标准物质是获得准确、可靠、具有可比性检测结果的保证。转基因大豆(Glycine max)MON89788为我国批准进口用作加工原料的农业转基因生物,因此,制备转基因大豆MON89788基体标准物质是对其进行安全管理所必需的。本研究通过将转基因大豆MON89788和其受体材料A3244分别冷冻研磨成粉末,按质量分数称量配制,得到转基因含量为49.3 g/kg的标准物质。采用方差分析法(F检验法)进行均匀性检验,统计量F值为1.618,小于临界值F_((0.05,11,24)),标准物质均匀性良好;短期稳定性考察结果表明,标准物质可在常温下稳定储存4周,在-20℃下长期稳定性达到30个月。本批MON89788基体标准物质有两个特性量值,一是基于重量法配比混合的粉末质量分数,量值为49.3 g/kg,扩展不确定度为2.0 g/kg;二是基于q RT-PCR 8家实验室联合定值的转基因DNA与总DNA的质量分数,量值为49.8 ng/μg,扩展不确定度为5.9 ng/μg。本批标准物质每瓶1.0 g,使用时最小取样量为100 mg,可用于对转基因大豆MON89788进行定性和定量检测,以及转基因大豆MON89788特异性检测方法的评价和实验室质量控制。本研究为我国转基因检测基体标准物质的研制提供了理论基础和技术支持。
Detection of genetically modified product components is an important support for GMO safety management and identification, and GMO standard material is to obtain accurate, reliable, comparable test results guarantee. Glycine max MON89788 is an agricultural genetically modified organism approved for import in China as a raw material for processing. Therefore, it is necessary to prepare GM substrate MON89788 standard substance for its safety management. In this study, transgenic soybean MON89788 and its receptor material A3244 were respectively cryo-ground into powder and weighed in mass fraction to obtain a standard substance with a transgene content of 49.3 g / kg. The variance test (F test) was used to test the uniformity. The statistic F value was 1.618, which was less than the critical value F _ ((0.05,11,24)), and the standard substance had good homogeneity. The short-term stability test showed that the standard substance Stable storage at room temperature for 4 weeks, long-term stability at -20 ℃ for 30 months. This batch of MON89788 matrix standard material has two characteristic values, one is based on gravimetric mixing powder mass fraction, the value of 49.3 g / kg, the expansion of uncertainty of 2.0 g / kg; the second is based on q RT- PCR 8 laboratories jointly set the mass fraction of transgenic DNA and total DNA, the value of 49.8 ng / μg, extended uncertainty of 5.9 ng / μg. This batch of standard material 1.0 g per bottle, the minimum sample size of 100 mg when used, can be used for qualitative and quantitative detection of transgenic soybean MON89788, as well as genetically modified soybean MON89788-specific evaluation methods and laboratory quality control. This study provided the theoretical basis and technical support for the development of our country’s standard substance for the detection of genetically modified organisms.