A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops
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 immunochromatogra...
Saved in:
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Tsinghua University Press
2024-01-01
|
| Series: | Food Science and Human Wellness |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213453023001325 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832557740957892608 |
|---|---|
| author | Jinbin Wang Yu Wang Xiuwen Hu Yifan Chen Wei Jiang Xiaofeng Liu Juan Liu Lemei Zhu Haijuan Zeng Hua Liu |
| author_facet | Jinbin Wang Yu Wang Xiuwen Hu Yifan Chen Wei Jiang Xiaofeng Liu Juan Liu Lemei Zhu Haijuan Zeng Hua Liu |
| author_sort | Jinbin Wang |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-426fff0968964f8a966c686448b16199 |
| institution | Kabale University |
| issn | 2213-4530 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | Tsinghua University Press |
| record_format | Article |
| series | Food Science and Human Wellness |
| spelling | doaj-art-426fff0968964f8a966c686448b161992025-02-03T02:51:42ZengTsinghua University PressFood Science and Human Wellness2213-45302024-01-01131183190A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified cropsJinbin Wang0Yu Wang1Xiuwen Hu2Yifan Chen3Wei Jiang4Xiaofeng Liu5Juan Liu6Lemei Zhu7Haijuan Zeng8Hua Liu9Key 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, ChinaKey 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, ChinaKey Laboratory of Agricultural Genetics and Breeding, Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Shanghai 201106, ChinaKey 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, ChinaKey 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, ChinaSchool of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, ChinaSchool of Public Health, Changsha Medical University, Changsha 410219, China; Academician Workstation, Changsha Medical University, Changsha 410219, ChinaSchool of Public Health, Changsha Medical University, Changsha 410219, China; Academician Workstation, Changsha Medical University, Changsha 410219, ChinaKey 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.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.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.http://www.sciencedirect.com/science/article/pii/S2213453023001325Genetically modified cropsOn-site detectionLateral flow test stripsDual recombinase polymerase amplification (RPA) |
| spellingShingle | Jinbin Wang Yu Wang Xiuwen Hu Yifan Chen Wei Jiang Xiaofeng Liu Juan Liu Lemei Zhu Haijuan Zeng Hua Liu 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 Genetically modified crops On-site detection Lateral flow test strips Dual recombinase polymerase amplification (RPA) |
| title | A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops |
| title_full | A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops |
| title_fullStr | A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops |
| title_full_unstemmed | A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops |
| title_short | A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops |
| title_sort | dual rpa based lateral flow strip for sensitive on site detection of cp4 epsps and cry1ab ac genes in genetically modified crops |
| topic | Genetically modified crops On-site detection Lateral flow test strips Dual recombinase polymerase amplification (RPA) |
| url | http://www.sciencedirect.com/science/article/pii/S2213453023001325 |
| work_keys_str_mv | AT jinbinwang adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT yuwang adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT xiuwenhu adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT yifanchen adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT weijiang adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT xiaofengliu adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT juanliu adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT lemeizhu adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT haijuanzeng adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT hualiu adualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT jinbinwang dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT yuwang dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT xiuwenhu dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT yifanchen dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT weijiang dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT xiaofengliu dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT juanliu dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT lemeizhu dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT haijuanzeng dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops AT hualiu dualrpabasedlateralflowstripforsensitiveonsitedetectionofcp4epspsandcry1abacgenesingeneticallymodifiedcrops |