Rapid detection of genetically modified products based on CRISPR-Cas12a combined with recombinase polymerase amplification
文献类型： 外文期刊
作者： Wang, Jinbin 1 ; Hu, Xiuwen 1 ; Wang, Yu 1 ; Zeng, Haijuan 1 ; Liu, Xiaofeng 4 ; Liu, Hua 1 ;
作者机构： 1.Inst Biotechnol Res, Shanghai Acad Agr Sci, Key Lab Agr Genet & Breeding, 2901 Beidi Rd, Shanghai 201106, Peoples R China
2.Minist Agr & Rural Affairs, Crops Ecol Environm Secur Inspection & Supervis Ct, RC,2901 Beidi Rd, Shanghai 201106, Peoples R China
3.Shanghai Oecean Univ, Coll Food Sci & Technol, 999 Huancheng Rd, Shanghai 200120, Peoples R China
4.Lanzhou Univ Technol, Sch Life Sci & Engn, Lanzhou 730050, Peoples R China
5.Shanghai Acad Agr Sci, Shanghai, Peoples R China
关键词： Genetically modified crops; CRISPR-Cas12a; CP4-EPSPS; Cry1Ab; Ac; Rapid detection
期刊名称：CURRENT RESEARCH IN FOOD SCIENCE （ 影响因子：6.3； 五年影响因子：6.3 ）
ISSN：
年卷期： 2022 年 5 卷
页码：
收录情况： SCI
摘要： 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/mu L of the standard plasmid con-taining 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.
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