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  • Title: Development and application of a quantitative loop-mediated isothermal amplification method for detecting genetically modified maize MON863.
    Author: Huang S, Xu Y, Yan X, Shang Y, Zhu P, Tian W, Xu W.
    Journal: J Sci Food Agric; 2015 Jan; 95(2):253-9. PubMed ID: 24771525.
    Abstract:
    BACKGROUND: A 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. RESULTS: The qualitative and quantitative reaction conditions (dNTPs, betaine, primers, Mg(2+), Bst polymerase, temperature, reaction time) were optimized. The concentrations of Mg(2+) 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 °C. CONCLUSION: The results of this study demonstrate that this new SYBR Green I-based quantitative LAMP assay system is reliable, sensitive and accurate.
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