RPA-Cas12a-FS: A frontline nucleic acid rapid detection system for food safety based on CRISPR-Cas12a combined with recombinase polymerase amplification
RPA-Cas12a-FS: A frontline nucleic acid rapid detection system for food safety based on CRISPR-Cas12a combined with recombinase polymerase amplification
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DOI：
10.1016/j.foodchem.2020.127608
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发表时间：
2021-01-01
期刊：
影响因子：
8.8
通讯作者：
Tang, Xueming
中科院分区：
文献类型：
--
作者：
Liu, Hua;Wang, Jinbin;Tang, Xueming
确保食品安全和质量的食品分析与所有国家都相关。本研究旨在开发一种将重组酶聚合酶扩增与CRISPR-Cas 12 a结合用于食品安全的检测技术（称为RPA-Cas 12 a-FS）。我们的数据表明，这种新的方法可以检测通过荧光强度的分子鉴定食源性致病菌，转基因作物和肉类掺假。优化后，RPA-Cas 12 a-FS的灵敏度和稳定性进一步提高。RPA-Cas 12 a-FS系统可以在37 ℃下在45分钟内特异性检测低至10个拷贝的靶基因水平。RPA-Cas 12 a-FS系统对实验室标准样品和现场样品均具有较高的灵敏度，表明该检测方法具有一定的实用性。综上所述，基于CRISPR-Cas 12 a的简单、快速、高灵敏度的检测方法被开发用于食品安全领域的分子鉴定，而不需要技术专业知识或辅助设备。
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 degrees 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.