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

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

外文关键词：

Genetically modified cropsOn-site detectionLateral flow test stripsDual recombinase polymerase amplification(RPA)

作者：

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

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作者单位：

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

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基金：

Scientific and Innovative Action Plan of ShanghaiShanghai Rising-Star ProgramShanghai Sailing ProgramShanghai Science and Technology Commission,the Belt and Road ProjectTalent Project of SAAS(2023-2025)Runup Plan of SAASSAAS Program for Excellent Research Team

项目编号：

21N3190080023QB140350020YF144300020310750500ZP222112022B-16

出版年：

2024

DOI：

10.26599/FSHW.2022.9250015

食品科学与人类健康(英文)

CSTPCD

ISSN：

年,卷(期)：2024.13(1)

参考文献量35