Liu Hua, Hu Xiuwen, Zeng Haijuan, He Chuan, Cheng Fang, Tang Xueming, Wang Jinbin
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 Ocean University, 999 Huancheng Road Shanghai, 200120, China.
Curr Res Food Sci. 2023 Oct 4;7:100605. doi: 10.1016/j.crfs.2023.100605. eCollection 2023.
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-CRISPR-Cas12a-LFA). It is an intuitive, sensitive and specific fluorescence detection and test strip system to detect and 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 °C. The system was used to evaluate field test samples and showed high reproducibility after testing products containing and 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.
随着全球转基因作物种植面积的不断增加，快速高效的检测技术对于转基因生物的监管和筛选变得非常重要。我们构建了一种基于环介导等温扩增（LAMP）、CRISPR-Cas12a和侧向流动分析（LAMP-CRISPR-Cas12a-LFA）的方法。它是一种直观、灵敏且特异的荧光检测和试纸条系统，用于田间筛选中检测和基因。在使用筛选出的特异性引物优化条件后，基于侧向流动试纸条，LAMP-CRISPR-Cas12a-LFA方法的检测限（LOD）为100拷贝，整个检测过程在61℃下1小时内即可完成。该系统用于评估田间测试样品，在检测含有和基因的产品后显示出高重现性，且两者均能被检测到。本文建立的LAMP-CRISPR-Cas12a-LFA方法作为一种快速的田间检测方法。它仅需一台便携式恒温仪器，这使其适用于田间样品的快速检测，并可在实验工作站、执法现场工作以及当地检验检疫部门使用。
