Rapid detection of genetically modified products based on CRISPR-Cas12a combined with recombinase polymerase amplification

Jinbin Wang; Xiuwen Hu; Yu Wang; Haijuan Zeng; Xiaofeng Liu; Hua Liu

Current Research in Food Science (Jan 2022)

Rapid detection of genetically modified products based on CRISPR-Cas12a combined with recombinase polymerase amplification

Jinbin Wang,
Xiuwen Hu,
Yu Wang,
Haijuan Zeng,
Xiaofeng Liu,
Hua Liu

Affiliations

Jinbin Wang: Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Key Laboratory of Agricultural Genetics and Breeding, 2901 Beidi Road, Shanghai, 201106, China; Crops Ecological Environment Security Inspection and Supervision Center (Shanghai), Ministry of Agriculture and Rural Affairs, P.R.C., 2901 Beidi Road, Shanghai, 201106, China; Corresponding author. Shanghai Academy of Agricultural Sciences, Shanghai, China.
Xiuwen Hu: Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Key Laboratory of Agricultural Genetics and Breeding, 2901 Beidi Road, Shanghai, 201106, China; College of Food Sciences and Technology, Shanghai Oecean University, 999 Huancheng Road, Shanghai, 200120, China
Yu Wang: Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Key Laboratory of Agricultural Genetics and Breeding, 2901 Beidi Road, Shanghai, 201106, China
Haijuan Zeng: Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Key Laboratory of Agricultural Genetics and Breeding, 2901 Beidi Road, Shanghai, 201106, China; Crops Ecological Environment Security Inspection and Supervision Center (Shanghai), Ministry of Agriculture and Rural Affairs, P.R.C., 2901 Beidi Road, Shanghai, 201106, China
Xiaofeng Liu: School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China
Hua Liu: Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Key Laboratory of Agricultural Genetics and Breeding, 2901 Beidi Road, Shanghai, 201106, China; Crops Ecological Environment Security Inspection and Supervision Center (Shanghai), Ministry of Agriculture and Rural Affairs, P.R.C., 2901 Beidi Road, Shanghai, 201106, China; Corresponding author. Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences; Key Laboratory of Agricultural Genetics and Breeding, 2901 Beidi Road, Shanghai, 201106, China.

Journal volume & issue: Vol. 5
pp. 2281 – 2286

Abstract

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With the large-scale planting of genetically modified (GM) crops, consumers were more aware of biosafety. Onsite rapid diagnostic methods were advantageous to the regulation of GM products. In this study, a rapid, sensitive and portable detection method based on recombinase polymerase amplification were proposed based on RPA reaction and Cas12a cleavage reaction for GM ingredients, named RPA-Cas12a-GM. The results would be displayed by fluorescence signal (FS) and visual bands of lateral flow strip (LFS). RPA-Cas12a-GM method could be completed within 45 min, and the detection limit was as low as 45 copies/μL of the standard plasmid containing CP4-EPSPS gene and Cry1Ab/Ac gene. Furthermore, the detection coincidence rate of RPA-Cas12a-GM method was 100%. In conclusion, the proposed RPA-Cas12a-GM method based FS and LFS were sensitive, specific, rapid and visible for diagnosis of CP4-EPSPS gene and Cry1Ab/Ac gene without complex equipment, which provides technical support for the regulation of GM products in the field.

Published in Current Research in Food Science

ISSN: 2665-9271 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Technology: Home economics: Nutrition. Foods and food supply; Technology: Chemical technology: Food processing and manufacture
Website: https://www.journals.elsevier.com/current-research-in-food-science

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