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Research Article | Open Access

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

Jinbin Wanga,b,1, Yu Wanga,c,1, Xiuwen Hua,, Yifan Chena,b, Wei Jianga,b, Xiaofeng Liuc, Juan Liud,e, Lemei Zhud,e, Haijuan Zenga,b( ), Hua Liua,b( )
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
School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China
School of Public Health, Changsha Medical University, Changsha 410219, China
Academician Workstation, Changsha Medical University, Changsha 410219, China

1 Jinbin Wang and Yu Wang contributed equally to this work.

Peer review under responsibility of Tsinghua University Press.

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Abstract

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 forCP4-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, equipmentfree field manner, providing a new alternative for rapid screening for transgenic assays in the field.

Keywords

On-site detectionLateral flow test stripsGenetically modified cropsDual recombinase polymerase amplification(RPA)

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Food Science and Human Wellness
Volume 13 Issue 1,
January 2024
Pages 183-190
DOI: 10.26599/FSHW.2022.9250015
Cite this article:
Wang J, Wang Y, Hu X, et al. A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops. Food Science and Human Wellness, 2024, 13(1): 183-190. https://doi.org/10.26599/FSHW.2022.9250015

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Received: 07 January 2022
Revised: 27 February 2022
Accepted: 30 March 2022
Published: 01 June 2023
© 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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