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首页> 外文期刊>Food Control >Loop-mediated isothermal amplification targeting insect resistant and herbicide tolerant transgenes: Monitoring for GM contamination in supply chain
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Loop-mediated isothermal amplification targeting insect resistant and herbicide tolerant transgenes: Monitoring for GM contamination in supply chain

机译：靶向昆虫抗性和除草剂耐受性转基因的环介导等温扩增：监测供应链中的转基因污染

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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, BM, Bt176, MON89034), cotton (MON531, MON15985, GFM-cry 1A, Event1, MLS9124, MON1445, MON15985 x MON88913), eggplant (EE1) and soybean (GTS40-3-2). Real-time LAMP was found sensitive enough to detect as low as 2 copies for crylAc 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. (C) 2014 Elsevier Ltd. All rights reserved.
机译：由于全球对转基因作物的采用前所未有地增加，因此需要有效的转基因作物检测策略来有效解决某些生物安全和释放后监测问题。除草剂耐受性和杀虫剂抗性是商业化的转基因粮食作物的主要特征。已经报道了基于环介导的等温扩增（LAMP）的视觉和实时检测系统，该系统靶向鳞翅目昆虫抗性cry1Ac，cry2Ab2和草甘膦耐受性cp4-epsps基因。使用四种作物的十四次转基因事件，即玉米（MON810，NK603，BM，Bt176，MON89034），棉花（MON531，MON15985，GFM-cry 1A，事件1，MLS9124，MON1445，MON15985 x MON88913）确认LAMP分析的特异性，茄子（EE1）和大豆（GTS40-3-2）。使用校准曲线，发现实时LAMP足够灵敏，足以在35分钟内检测出cry1Ac低至2拷贝，cry2Ab2和cp4-epsps低至4拷贝。视觉LAMP分析的检测限（LOD）降至0.01％（4份GM含量），低于常规PCR（根据目标检测40-400份）。 LAMP分析比常规PCR更快，更方便用户使用，可高效，高特异性和高灵敏度地用于监测食品和饲料供应链中的转基因污染。结合快速DNA提取方法开发的检测方法，将有助于现场检测，以检查进入口和农民田地中样品或产品的GM状态。 （C）2014 Elsevier Ltd.保留所有权利。

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