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Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification

文献类型： 外文期刊

作者： Chen, Xiaoyun 1 ; Wang, Xiaofu 1 ; Jin, Nuo 2 ; Zhou, Yu 1 ; Huang, Sainan 1 ; Miao, Qingmei 1 ; Zhu, Qing 1 ; Xu, Junfen 1 ;

作者机构： 1.Zhejiang Acad Agr Sci, Inst Agr Qual & Stand Agroprod, Hangzhou 310021, Zhejiang, Peoples R China

2.E China Univ Sci & Technol, Sch Biotechnol, Shanghai 200237, Peoples R China

3.Shenyang Normal Univ, Coll Chem & Life Sci, Shenyang 110034, Peoples R China

关键词： GMO detection;LAMP;transgenic rice;TT51-1;KMD1;KF6

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

ISSN： 1422-0067

年卷期： 2012 年 13 卷 11 期

页码：

收录情况： SCI

摘要： Genetically modified (GM) rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR), currently the most common method for detecting genetically modified organisms, requires temperature cycling and relatively complex procedures. Here we developed a visual and rapid loop-mediated isothermal amplification (LAMP) method to amplify three GM rice event-specific junction sequences. Target DNA was amplified and visualized by two indicators (SYBR green or hydroxy naphthol blue [HNB]) within 60 min at an isothermal temperature of 63 degrees C. Different kinds of plants were selected to ensure the specificity of detection and the results of the non-target samples were negative, indicating that the primer sets for the three GM rice varieties had good levels of specificity. The sensitivity of LAMP, with detection limits at low concentration levels (0.01%-0.005% GM), was 10- to 100-fold greater than that of conventional PCR. Additionally, the LAMP assay coupled with an indicator (SYBR green or HNB) facilitated analysis. These findings revealed that the rapid detection method was suitable as a simple field-based test to determine the status of GM crops.

  • 相关文献

[1]Multiplex event-specific qualitative polymerase chain reaction for detecting three transgenic rice lines and application of a standard plasmid as a quantitative reference molecule. Wang, Xiaofu,Shen, Wenbiao,Wang, Xiaofu,Chen, Xiaoyun,Xu, Junfeng,Wang, Pengfei.

[2]Degradation and detection of transgenic Bacillus thuringiensis DNA and proteins in flour of three genetically modified rice events submitted to a set of thermal processes. Wang, Xiaofu,Chen, Xiaoyun,Xu, Junfeng,Wang, Xiaofu,Dai, Chen,Shen, Wenbiao.

[3]快速检测兔支气管败血波氏杆菌环介导等温扩增(LAMP)方法的建立及应用. 李科,刘燕,肖琛闻,韦强,鲍国连,季权安. 2013

[4]Ectopic expression of the spike protein of Rice Ragged Stunt Oryzavirus in transgenic rice plants inhibits transmission of the virus to insects. Shao, CG,Wu, JH,Zhou, GY,Sun, G,Peng, BZ,Lei, JL,Jin, DD,Chen, SX,Upadhyaya, NM,Waterhouse, P,Gong, ZX. 2003

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[6]Effectiveness in the field of Bt rice lines against target pests under various cultural regimes. Yang, Yajun,Xu, Hongxing,Zheng, Xusong,Tian, Junce,Lu, Zhongxian,Han, Hailiang,Wang, Guiyue,Lin, Yongjun,Lin, Yongjun.

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