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利用侧流层析试纸环介导等温扩增快速筛选检测转基因作物。

Rapid Screening Detection of Genetically Modified Crops by Loop-Mediated Isothermal Amplification with a Lateral Flow Dipstick.

机构信息

Division of Analytical Science, Food Research Institute , National Agriculture and Food Research Organization , 2-1-12 Kannondai , Tsukuba , Ibaraki 305-8642 , Japan.

FASMAC Co., Ltd. , 5-1-3 Midorigaoka , Atsugi , Kanagawa 243-0041 , Japan.

出版信息

J Agric Food Chem. 2018 Jul 25;66(29):7839-7845. doi: 10.1021/acs.jafc.8b01765. Epub 2018 Jul 11.

Abstract

We developed a novel loop-mediated isothermal amplification (LAMP)-based detection method using lateral flow dipstick chromatography for genetically modified (GM) soybean and maize events. The single-stranded tag hybridization (STH) for the chromatography printed-array strip (C-PAS) system was used for detections targeting the cauliflower mosaic virus 35S promoter, mannose-6-phosphate isomerase gene, Pisum sativum ribulose 1, 5-bisphosphate carboxylase terminator, a common sequence between the Cry1Ab and Cry1Ac genes, and a GA21-specific sequence. The STH C-PAS system was applicable for multiplex analyses to perform simultaneous detections. The limit of detection was 0.5% or less for each target. By using the developed method, the LAMP amplification was visually detected. Moreover, the detection could be carried out without any expensive instruments, even for the DNA amplification steps, by virtue of the isothermal reaction. We demonstrated that the rapid and useful method developed here would be applicable for screening GM crops.

摘要

我们开发了一种基于环介导等温扩增(LAMP)的新型检测方法,该方法使用侧流层析试纸条(C-PAS)进行基因改良(GM)大豆和玉米事件的检测。针对花椰菜花叶病毒 35S 启动子、甘露糖-6-磷酸异构酶基因、豌豆 RuBisCO 小亚基终止子、Cry1Ab 和 Cry1Ac 基因之间的共同序列以及 GA21 特异性序列,我们使用单链标签杂交(STH)进行了 C-PAS 系统的检测。STH C-PAS 系统适用于多重分析,可同时进行检测。每个目标的检测限为 0.5%或更低。通过使用所开发的方法,可以对 LAMP 扩增进行直观检测。此外,由于等温反应,即使对于 DNA 扩增步骤,也可以无需任何昂贵的仪器进行检测。我们证明,这里开发的快速有用的方法将适用于 GM 作物的筛选。

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