A rapid and high-throughput system for the detection of transgenic products based on LAMP-CRISPR-Cas12a
文献类型： 外文期刊
作者： Liu, Hua 1 ; Hu, Xiuwen 1 ; Zeng, Haijuan 1 ; He, Chuan 1 ; Cheng, Fang 2 ; Tang, Xueming 3 ; Wang, Jinbin 1 ;
作者机构： 1.Shanghai Acad Agr Sci, Inst Biotechnol Res, Key Lab Agr Genet & Breeding, 2901 Beidi Rd, Shanghai 201106, Peoples R China
2.Shanghai Ocean Univ, Coll Food Sci & Technol, 999 Huancheng Rd, Shanghai 200120, Peoples R China
3.Shanghai Jiao Tong Univ, Inst Nat Sci, Shanghai 200240, Peoples R China
4.Shanghai Jiao Tong Univ, Sch Agr & Biol, Shanghai 200240, Peoples R China
关键词： GM organisms; CRISPR-Cas12a; LAMP; 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： 2023 年 7 卷
页码：
收录情况： SCI
摘要： With the increasing acreage of genetically modified crops worldwide, rapid and efficient detection technologies have become very important for the regulation and screening of GM organisms. We constructed a method based on loop-mediated isothermal amplification (LAMP), CRISPR-Cas12a and lateral flow assay (LAMP-CRISPRCas12a-LFA). It is an intuitive, sensitive and specific fluorescence detection and test strip system to detect CP4EPSPS and Cry1Ab/Ac genes in field screening. The LAMP-CRISPR-Cas12a-LFA method has a limit of detection (LOD) of 100 copies based on lateral flow test strips after optimization of the conditions with screened specific primers, and the entire detection process can be completed within 1 h at 61 degrees C. The system was used to evaluate field test samples and showed high reproducibility after testing products containing CP4-EPSPS and Cry1Ab/Ac genes, and both were detectable. The LAMP-CRISPR-Cas12a-LFA method established in this paper functions as a rapid field detection method. It requires only one portable thermostatic instrument, which renders it compatible with the rapid detection of field samples and useable at experimental workstations, in law enforcement field work, and in local inspection and quarantine departments.
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