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.
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.
Food Chem. 2021 Jan 1;334:127608. doi: 10.1016/j.foodchem.2020.127608. Epub 2020 Jul 19.
Food analysis to ensure food safety and quality are relevant to all countries. This study aimed to develop a detection technique by combining recombinase polymerase amplification with CRISPR-Cas12a for food safety (termed RPA-Cas12a-FS). Our data showed that this novel method could be detected via fluorescence intensity for the molecular identification of foodborne pathogenic bacteria, genetically modified crops, and meat adulteration. After optimization, the sensitivity and stability of RPA-Cas12a-FS was further enhanced. The RPA-Cas12a-FS system could specifically detect target gene levels as low as 10 copies in 45 min at 37 °C. The RPA-Cas12a-FS system was sensitive both using standard samples in the lab and using samples from the field, which indicated that this detection method was practical. In conclusion, a simple, rapid, and highly sensitive detection method based on CRISPR-Cas12a was developed for molecular identification in the food safety field without requiring technical expertise or ancillary equipment.
食品分析对于确保食品安全和质量至关重要，这一点对所有国家都适用。本研究旨在开发一种检测技术，将重组酶聚合酶扩增与 CRISPR-Cas12a 相结合用于食品安全检测（称为 RPA-Cas12a-FS）。我们的数据表明，这种新方法可以通过荧光强度进行检测，用于食源性致病菌、转基因作物和肉类掺假的分子鉴定。经过优化，RPA-Cas12a-FS 的灵敏度和稳定性得到进一步提高。RPA-Cas12a-FS 系统在 37°C 下 45 分钟内即可特异性检测到低至 10 个拷贝的靶基因水平。RPA-Cas12a-FS 系统在实验室标准样本和现场样本中的检测均具有敏感性，这表明该检测方法具有实用性。总之，开发了一种基于 CRISPR-Cas12a 的简单、快速、高灵敏度的检测方法，用于食品安全领域的分子鉴定，无需专业技术或辅助设备。
