Development of the Visual Loop-Mediated Isothermal Amplification Assays for Seven Genetically Modified Maize Events and Their Application in Practical Samples Analysis

被引:50
作者
Chen, Lili [1 ,2 ]
Guo, Jinchao [1 ]
Wang, Qidi [1 ]
Kai, Guoyin [2 ]
Yang, Litao [1 ]
机构
[1] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, SJTU Bor Luh Food Safety Ctr, Natl Ctr Mol Characterizat Genetically Modified O, Shanghai 200240, Peoples R China
[2] Shanghai Normal Univ, Coll Life & Environm Sci, Plant Biotechnol Lab, Shanghai 200234, Peoples R China
关键词
Genetically modified maize; loop-mediated isothermal amplification; event-specific; RAPID DETECTION; PCR; QUANTIFICATION; TECHNOLOGY; LIGATION; SIMPLEX; GMOS; DNA;
D O I
10.1021/jf200459s
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
As more and more genetically modified (GM) crops are approved for commercialization and planting, the development of quick and on-spot methods for GM crops and their derivates is required. Herein, we established the polymerase chain reaction and agarose gel electrophoresis-free system for the identification of seven GM maize events (DAS-59122-7, T25, BT176, TC1507, MON810, BT11, and MON863) employing a loop-mediated isothermal amplification (LAMP) technique. The LAMP assay was performed using a set of four specific primers at 60-65 degrees C in less than 40 min, and the results were observed by direct visual observation. In these developed assays, the specificity targeted at each GM maize event based on the event-specific sequence was well confirmed, and the limits of detection were as low as four copies of maize haploid genomic DNA with an exception of 40 copies for MON810 assay. Furthermore, these developed assays were successfully used to test six practical samples with different GM maize events and contents (ranged from 0.0 to 2.0%). All of the results indicated that the established event-specific visual LAMP assays are more convenient, rapid, and low-cost for GM maize routine analysis.
引用
收藏
页码:5914 / 5918
页数:5
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