Development and application of a quantitative loop‐mediated isothermal amplification method for detecting genetically modified maize MON863. (27th May 2014)

• Record Type: Journal Article
• Authors: Huang, Sicong; Xu, Yuancong; Yan, Xinghua; Shang, Ying; Zhu, Pengyu; Tian, Wenying; Xu, Wentao
• Is Part Of: Journal of the science of food and agriculture. Volume 95:Number 2(2015:Jan. 30)
• Page Start: 253
• Page End: 259
• Publication Date: 2014-05-27
• Subjects: Food -- Periodicals; Agriculture -- Periodicals; 664
• DOI: 10.1002/jsfa.6707
• Languages: English
• ISSNs: 0022-5142

Abstract

<abstract abstract-type="main" id="jsfa6707-abs-0001"> <title>Abstract</title> <sec id="jsfa6707-sec-0001" sec-type="section"> <title>BACKGROUND</title> <p id="jsfa6707-para-0001">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 <italic>Cry3Bb1</italic> and <italic>tahsp17</italic> genes.</p> </sec> <sec id="jsfa6707-sec-0002" sec-type="section"> <title>RESULTS</title> <p id="jsfa6707-para-0002">The 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 °C.</p> </sec> <sec id="jsfa6707-sec-0003" sec-type="section"> <title>CONCLUSION</title> <p id="jsfa6707-para-0003">The results of this study demonstrate that this new SYBR Green I‐based quantitative LAMP assay system is reliable, sensitive and accurate. © 2014 Society of Chemical Industry</p> </sec> </abstract>