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RPA-Cas12a-FS: A frontline nucleic acid rapid detection system for food safety based on CRISPR-Cas12a combined with recombinase polymerase amplification

文献类型： 外文期刊

作者： Liu, Hua 1 ; Wang, Jinbin 1 ; Zeng, Haijuan 1 ; Liu, Xiaofeng 4 ; Jiang, Wei 1 ; Wang, Yu 4 ; Ouyang, Wanbao 4 ; Tang, Xu 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Inst Biotechnol Res, 2901 Beidi Rd, Shanghai 201106, Peoples R China

2.Key Lab Agr Genet & Breeding, 2901 Beidi Rd, Shanghai 201106, Peoples R China

3.Minist Agr & Rural Affairs, Crops Ecol Environm Secur Inspect & Supervis Ctr, 2901 Beidi Rd, Shanghai 201106, Peoples R China

4.Lanzhou Univ Technol, Sch Life Sci & Engn, Lanzhou 730050, Peoples R China

关键词： CRISPR-Cas12a; Detection; Foodborne pathogenic bacteria; Genetically modified crops; Meat adulteration

期刊名称：FOOD CHEMISTRY （ 影响因子：7.514； 五年影响因子：7.516 ）

ISSN： 0308-8146

年卷期： 2021 年 334 卷

页码：

收录情况： SCI

摘要： 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.

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