转基因玉米T25数字PCR方法的建立与验证

doi:10.13304/j.nykjdb.2019.0154

中国农业科技导报 ›› 2020, Vol. 22 ›› Issue (2): 173-178.DOI: 10.13304/j.nykjdb.2019.0154

转基因玉米T25数字PCR方法的建立与验证

李夏莹1,武玉花2,李俊2,肖晓琳1,张飞燕1,梁晋刚1,王顥潜1,张旭冬1,张秀杰1*

1.农业农村部科技发展中心, 北京 100176; 2.中国农业科学院油料作物研究所, 武汉 430062

收稿日期:2019-03-07 出版日期:2020-02-15 发布日期:2019-03-28
通讯作者: *通信作者张秀杰 E-mail:zhxj7410@sina.com
作者简介:李夏莹 E-mail: esmacloed006@163.com;
基金资助:
国家科技重大专项（2016ZX08012003）。

Establishment and Testing of Genetically Modified T25 Maize Digital PCR Method

LI Xiaying1, WU Yuhua2, LI Jun2, XIAO Xiaolin1, ZHANG Feiyan1, LIANG Jingang1, WANG Haoqian1, ZHANG Xudong1, ZHANG Xiujie1*

1.Development Center for Science and Technology, Ministry of Agriculture and Rural Affairs, Beijing 100176, China; 2.Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, China

Received:2019-03-07 Online:2020-02-15 Published:2019-03-28

摘要/Abstract

摘要： 转基因安全管理和标识制度的实施需要标准化的检测方法和转基因检测标准物质，标准物质是获得准确、可靠、可比检测结果的保证。转基因玉米T25为我国批准进口用作加工原料的转基因植物。为加强对T25的监管，以T25基体标准物质为研究对象，建立数字PCR方法，并选择8家实验室，采用数字PCR联合定值测定。结果表明，T25/Adh1二重数字PCR系统具有良好的扩增特异性。初始模板量在100～10 000拷贝·μL-1之间可获得稳定、可靠的检测结果。通过多实验室协同定值说明T25/Adh1二重数字PCR方法准确、可靠。T25基体标准物质的量值为1.001 2，相对不确定度为0.001 6。研究表明建立的T25/Adh1二重数字PCR方法可以用于转基因玉米T25的定量检测，为转基因成分定量检测提供了物质基础和技术保证。

关键词: 转基因玉米, T25, 基体标准物质, 微滴数字PCR

Abstract: The implementation of genetically modified organism (GMO) safety management and labeling system requires standardized testing methods and GMO testing standard substances, which guarantee the acquirement of accurate, reliable and comparable testing results. GMO maize T25 was approved for import as processing raw material in China. In order to strengthen the supervision of T25, the present work took the T25 matrix standard substance as the research object and established the digital PCR method. Eight laboratories were selected and the digital PCR combined quantitative determination were adopted. The results showed that the T25/Adh1 dual digital PCR system had good amplification specificity. Stable and reliable detection results could be obtained between 100 and 10 000 copies·μL-1 of the initial template. The method of T25/Adh1 dual digital PCR was proved to be accurate and reliable through multi-laboratory collaborative evaluation. The quantitative value of T25 matrix standard substance was 1.001 2 and the relative uncertainty was 0.001 6. It was established that T25/Adh1 dual digital PCR method could be used for the quantitative evaluation of genetically modified corn T25, which provided material basis and technical guarantee for the quantitative detection of genetically modified components.

Key words: genetically modified maize, T25, matrix reference material, microdrop digital PCR

李夏莹1,武玉花2,李俊2,肖晓琳1,张飞燕1,梁晋刚1,王顥潜1,张旭冬1,张秀杰1*. 转基因玉米T25数字PCR方法的建立与验证[J]. 中国农业科技导报, 2020, 22(2): 173-178.

LI Xiaying1, WU Yuhua2, LI Jun2, XIAO Xiaolin1, ZHANG Feiyan1, LIANG Jingang1, WANG Haoqian1, ZHANG Xudong1, ZHANG Xiujie1*. Establishment and Testing of Genetically Modified T25 Maize Digital PCR Method[J]. Journal of Agricultural Science and Technology, 2020, 22(2): 173-178.

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转基因玉米T25数字PCR方法的建立与验证

Establishment and Testing of Genetically Modified T25 Maize Digital PCR Method

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