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

被引:29
作者
Takabatake, Reona [1 ]
Kagiya, Yukari [2 ]
Minegish, Yasutaka [3 ]
Futo, Satoshi [2 ]
Soga, Keisuke [4 ]
Nakamura, Kosuke [4 ]
Kondo, Kazunari [4 ]
Mano, Junichi [1 ]
Kitta, Kazumi [1 ]
机构
[1] Natl Agr & Food Res Org, Food Res Inst, Div Analyt Sci, 2-1-12 Kannondai, Tsukuba, Ibaraki 3058642, Japan
[2] FASMAC Co Ltd, 5-1-3 Midorigaoka, Atsugi, Kanagawa 2430041, Japan
[3] Nippon Gene Co Ltd, Chiyoda Ku, 1-5 Kandanishiki Cho, Tokyo 1010054, Japan
[4] Natl Inst Hlth Sci, Kawasaki Ku, 3-25-26 Tonomachi, Kawasaki, Kanagawa 2109501, Japan
关键词
loop-mediated isothermal amplification (LAMP); genetically modified (GM); dipstick DNA chromatography; rapid; multiplex analysis; MODIFIED MAIZE; LABORATORY VALIDATION; MODIFIED ORGANISMS; VISUAL DETECTION; GMO DETECTION; FEED; GENE; FOOD;
D O I
10.1021/acs.jafc.8b01765
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
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 CrylAb 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.
引用
收藏
页码:7839 / 7845
页数:7
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