A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops
文献类型： 外文期刊
作者： Wang, Jinbin 1 ; Wang, Yu 1 ; Hu, Xiuwen 1 ; Chen, Yifan 1 ; Jiang, Wei 1 ; Liu, Xiaofeng 3 ; Liu, Juan 4 ; Zhu, Lemei 4 ; Zeng, Haijuan 1 ; Liu, Hua 1 ;
作者机构： 1.Shanghai Acad Agr Sci, Inst Biotechnol Res, Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China
2.Minist Agr & Rural Affairs, Crops Ecol Environm Secur Inspect & Supervis Ctr S, Shanghai 201106, Peoples R China
3.Lanzhou Univ Technol, Sch Life Sci & Engn, Lanzhou 730050, Peoples R China
4.Changsha Med Univ, Sch Publ Hlth, Changsha 410219, Peoples R China
5.Changsha Med Univ, Acad Workstat, Changsha 410219, Peoples R China
6.Shanghai Acad Agr Sci, Inst Biotechnol Res, Shanghai 201106, Peoples R China
关键词： Genetically modified crops; On-site detection; Lateral flow test strips; Dual recombinase polymerase amplification; (RPA)
期刊名称：FOOD SCIENCE AND HUMAN WELLNESS （ 影响因子：7.0； 五年影响因子：8.3 ）
ISSN：
年卷期： 2024 年 13 卷 1 期
页码：
收录情况： SCI
摘要： Traditional transgenic detection methods require high test conditions and struggle to be both sensitive and efficient. In this study, a one-tube dual recombinase polymerase amplification (RPA) reaction system for CP4-EPSPS and Cry1Ab/Ac was proposed and combined with a lateral flow immunochromatographic assay, named "Dual-RPA-LFD", to visualize the dual detection of genetically modified (GM) crops. In which, the herbicide tolerance gene CP4-EPSPS and the insect resistance gene Cry1Ab/Ac were selected as targets taking into account the current status of the most widespread application of insect resistance and herbicide tolerance traits and their stacked traits. Gradient diluted plasmids, transgenic standards, and actual samples were used as templates to conduct sensitivity, specificity, and practicality assays, respectively. The constructed method achieved the visual detection of plasmid at levels as low as 100 copies, demonstrating its high sensitivity. In addition, good applicability to transgenic samples was observed, with no cross-interference between two test lines and no influence from other genes. In conclusion, this strategy achieved the expected purpose of simultaneous detection of the two popular targets in GM crops within 20 min at 37 & DEG;C in a rapid, equipmentfree field manner, providing a new alternative for rapid screening for transgenic assays in the field. & COPY; 2024 Beijing Academy of Food Sciences. Publishing services by Tsinghua University Press. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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