转基因大豆MON89788基体标准物质的研制

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摘要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qRT-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。

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关键词 ：转基因大豆, MON89788, 基体标准物质, 定值, qRT-PCR

Abstract：Genetically modified organism (GMO) ingredients detection is an important part of GMO safety management and GMO labeling, GMO detection reference material (RM) is guaranteed to get accurate, reliable, comparable GMO ingredients detection results. GM soybean (Glycine max) MON89788 is a GM plant that has been approved to import and use as in-process raw material in China. Therefore, preparation of GM soybean MON89788 RM is essential for its safety management. In this study, GM soybean MON89788 and traditional soybean seeds A3244 were milled using a cryo-grinding vibrating mill separately, then the GM soybean MON89788 matrix RM was prepared by weighting and mixing of GM and Non-GM powder. The value of MON89788 matrix RM was 49.3 g/kg in mass fraction. The mixture was further dispensed into brown bottles. Analysis of variance (F test) was performed to test homogeneity of RMs. The calculated F-statistics is 1.618, less than the critical value F (0.05, 11, 24). The results indicated that there was no significant difference between bottles and within bottles, and the RMs were homogeneous. Stability study results indicated the RMs could be stored under ambient conditions for 4 weeks, and stored at - 20 ℃ for 30 months or more. The certified value was assigned based on mass fraction of GMO powder and non-GMO powder with the units of g/kg, and mass fraction of GMO DNA and non-GMO DNA with the units of ng/μg. The certified value was expressed as (49.3±2.0) g/kg, and (49.8±5.9) ng/μg. This batch of RMs was 1.0 g/bottle, and the recommended minimum sample intake was 100 mg. The RMs could be used in qualitative and quantitative detection of GM soybean MON89788, as well as in standardization of GMO detection method and laboratory quality control. This work provides theoretical basis and technical support for the development of GM detection matrix RMs.

Key words： Genetically modified soybean MON89788 Matrix reference material Value characterization qRT-PCR

收稿日期: 2015-07-27 出版日期: 2015-12-29

基金资助:国家转基因生物新品种培育重大专项;国家自然科学基金资助项目;中南民族大学中央高校基本科研业务费专项

通讯作者: 吴刚 E-mail: wugang1976@gmail.com;wugang@caas.cn

引用本文:

张丽宋贵文沈平李文龙武玉花吴刚. 转基因大豆MON89788基体标准物质的研制[J]. , 2016, 24(2): 295-304.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2016/V24/I2/295

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