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Efficient detection strategies for genetically modified (GM) crops are required to effectively address some of the biosafety and post-release monitoring issues, as global adoption of GM crops has been unprecedently increased. Herbicide tolerance and insecticide resistance are the major traits in commercialized GM food crops. Visual as well as real-time detection system based on loop-mediated isothermal amplification (LAMP) targeting lepidopteron insect resistant cry1Ac, cry2Ab2 and glyphosate tolerant cp4-epsps genes has been reported. Specificity of LAMP assays were confirmed using fourteen GM events of four crops, namely, corn (MON810, NK603, Bt11, Bt176, MON89034), cotton (MON531, MON15985, GFM-cry1A, Event1, MLS9124, MON1445, MON15985 × MON88913), eggplant (EE1) and soybean (GTS40-3-2). Real-time LAMP was found sensitive enough to detect as low as 2 copies for cry1Ac and 4 copies for cry2Ab2 and cp4-epsps within 35 min using a calibration curve. The limit of detection (LOD) of visual LAMP assays was down to 0.01% (4 copies of GM content) which is lower than conventional PCR (detecting 40–400 copies depending on target). LAMP assays are faster and more user-friendly than conventional PCR and could be efficiently utilized for monitoring of GM contamination in food and feed supply chain, with high specificity and sensitivity. The developed assays, when combined with a fast DNA extraction method, will facilitate on-site detection to check the GM status of a sample or product at ports of entry and in farmers' fields. |
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