WANG Yong,LAN Qing-kuo,ZHU Zhu,et al.Development of Visual Loop-Mediate Isothermal Amplification Assay for Herbicide-resistant Soybean GTS40-3-2 and Its Derivates[J].Soybean Science,2014,33(04):570-573.[doi:10.11861/j.issn.1000-9841.2014.04.0570]
转基因大豆GTS40-3-2成分现场可视化检测方法的建立
- Title:
- Development of Visual Loop-Mediate Isothermal Amplification Assay for Herbicide-resistant Soybean GTS40-3-2 and Its Derivates
- 关键词:
- 转基因大豆GTS40-3-2; 转化体特异性; 环状等温扩增技术; 现场可视化检测
- Keywords:
- Herbicide-resistant soybean GTS40-3-2; Event specific; Loop-mediated isothermal amplification; Visualization technology
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 转基因大豆GTS40-3-2转化体是种植面积最广的转基因大豆品种,根据GTS40-3-2转化体特异性序列设计引物,应用可视化环介导等温扩增技术,对其进行快速扩增及可视化判断;同时建立转基因成分人工污染的食品模型,比较了LAMP法与定性PCR方法检测的敏感性。结果表明:该方法仅对GTS40-3-2转化体产生特异性扩增,灵敏度高达0.001%。所建立的GTS40-3-2转化体特异性现场可视化筛查方法具有较高的特异性与灵敏性,操作简单、快速,可用于转基因大豆GTS40-3-2成分污染的现场可视化检测。
- Abstract:
- It is important to detect herbicide-resistant soybean GTS40-3-2 and its derivates in food or food materials in order to prevent the illegal diffusion.A rapid visual loop-mediated isothermal amplification(LAMP)method assay for GTS40-3-2 event-specific detection was developed.A set of primers were designed according to the nucleotide sequence of the target,8 genetically modified organisms(GMO)were detected by LAMP for method specificity,and meanwhile the mode of artificially contaminated food was constructed to evaluate the sensitivity of LAMP assay and qualitative PCR method.The results showed that the GTS40-3-2 event had specific amplification,but the non-CpTI GMO submitted negative reactions.Sensitivity of LAMP assay for GTS40-3-2 was 0.001%.In conclusion,the LAMP assay developed in the present study is a specific,sensitive,simple and convenient method for the rapid screening of GTS40-3-2 in contaminated foods.
参考文献/References:
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备注/Memo
基金项目:农业科技成果转化资金项目(2011GB2A100011);科技部国际合作项目(2006DFA32380)。