Development and application of a quantitative loop-mediated isothermal amplification method for detecting genetically modified maize MON863

作者：Huang, Sicong; Xu, Yuancong; Yan, Xinghua; Shang, Ying; Zhu, Pengyu; Tian, Wenying; Xu, Wentao^*

来源：Journal of the Science of Food and Agriculture, 2015, 95(2): 253-259.

DOI：10.1002/jsfa.6707

• loop-mediated isothermal amplification
• quantitative
• genetically modified maize
• Cry3Bb1

BACKGROUNDA SYBR Green I-based quantitative loop-mediated isothermal amplification (LAMP) assay was developed for the rapid detection of genetically modified maize MON863. A set of primers was designed based on the integration region of the Cry3Bb1 and tahsp17 genes.
RESULTSThe qualitative and quantitative reaction conditions (dNTPs, betaine, primers, Mg2+, Bst polymerase, temperature, reaction time) were optimized. The concentrations of Mg2+ and betaine were found to be important to the LAMP assay. The detection limits of both qualitative and quantitative LAMP for MON863 were as low as 4 haploid genomic DNA, and the LAMP reactions can be completed within 1 h at an isothermal temperature of 65 degrees C.
CONCLUSIONThe results of this study demonstrate that this new SYBR Green I-based quantitative LAMP assay system is reliable, sensitive and accurate.

• 出版日期2015-1-30
• 单位清华大学; 中国农业大学