A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops

Jinbin Wang; Yu Wang; Xiuwen Hu; Yifan Chen; Wei Jiang; Xiaofeng Liu; Juan Liu; Lemei Zhu; Haijuan Zeng; Hua Liu

Food Science and Human Wellness (Jan 2024)

A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops

Jinbin Wang,
Yu Wang,
Xiuwen Hu,
Yifan Chen,
Wei Jiang,
Xiaofeng Liu,
Juan Liu,
Lemei Zhu,
Haijuan Zeng,
Hua Liu

Affiliations

Jinbin Wang: Key Laboratory of Agricultural Genetics and Breeding, Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; Crops Ecological Environment Security Inspection and Supervision Center (Shanghai), Ministry of Agriculture and Rural Affairs, Shanghai 201106, China
Yu Wang: Key Laboratory of Agricultural Genetics and Breeding, Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China
Xiuwen Hu: Key Laboratory of Agricultural Genetics and Breeding, Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China
Yifan Chen: Key Laboratory of Agricultural Genetics and Breeding, Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; Crops Ecological Environment Security Inspection and Supervision Center (Shanghai), Ministry of Agriculture and Rural Affairs, Shanghai 201106, China
Wei Jiang: Key Laboratory of Agricultural Genetics and Breeding, Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; Crops Ecological Environment Security Inspection and Supervision Center (Shanghai), Ministry of Agriculture and Rural Affairs, Shanghai 201106, China
Xiaofeng Liu: School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China
Juan Liu: School of Public Health, Changsha Medical University, Changsha 410219, China; Academician Workstation, Changsha Medical University, Changsha 410219, China
Lemei Zhu: School of Public Health, Changsha Medical University, Changsha 410219, China; Academician Workstation, Changsha Medical University, Changsha 410219, China
Haijuan Zeng: Key Laboratory of Agricultural Genetics and Breeding, Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; Crops Ecological Environment Security Inspection and Supervision Center (Shanghai), Ministry of Agriculture and Rural Affairs, Shanghai 201106, China; Corresponding author at: Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China.
Hua Liu: Key Laboratory of Agricultural Genetics and Breeding, Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; Crops Ecological Environment Security Inspection and Supervision Center (Shanghai), Ministry of Agriculture and Rural Affairs, Shanghai 201106, China; Corresponding author at: Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China.

Journal volume & issue: Vol. 13, no. 1
pp. 183 – 190

Abstract

Read online

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 ℃ in a rapid, equipment-free field manner, providing a new alternative for rapid screening for transgenic assays in the field.

Published in Food Science and Human Wellness

ISSN: 2213-4530 (Online)
Publisher: Tsinghua University Press
Country of publisher: China
LCC subjects: Technology: Home economics: Nutrition. Foods and food supply
Website: https://www.sciopen.com/journal/2097-0765

About the journal

Abstract

Keywords