CRISPR/Cas12a-Assisted Dual Visualized Detection of SARS-CoV-2 on Frozen Shrimps

文献类型： 外文期刊

第一作者： Qian, Siwenjie

作者： Qian, Siwenjie;Chen, Yanju;Wu, Jian;Ye, Zhangying;Wang, Xiaofu;Xu, Junfeng;Wang, Tingzhang;Che, Yang;Wu, Jian

作者机构：

关键词： SARS-CoV-2; food safety; dual detection; CRISPR; Cas12a; anti-contamination

期刊名称：BIOSENSORS-BASEL （ 影响因子：5.4； 五年影响因子：5.7 ）

ISSN：

年卷期： 2023 年 13 卷 1 期

页码：

收录情况： SCI

摘要： Given the possibility that food contaminated with SARS-CoV-2 might become an infection source, there is an urgent need for us to develop a rapid and accurate nucleic acid detection method for SARS-CoV-2 in food to ensure food safety. Here, we propose a sensitive, specific, and reliable molecular detection method for SARS-CoV-2. It has a mechanism to control amplicon contamination. Swabs from spiked frozen shrimps were used as detection samples, which were processed by heating at 95 degrees C for 30 s. These preprocessed samples served as the templates for subsequent amplification. A colorimetric LAMP reaction was carried out to amplify both the SARS-CoV-2 target and the MS2 phage simultaneously in one tube. MS2 phage was detected by colorimetric LAMP as the internal control, while SARS-CoV-2 was detected with a CRISPR/Cas12a system. The fluorescence results could be visually detected with an ultraviolet lamp. Meanwhile, uracil was incorporated during the LAMP reaction to provide an amplicon contamination proof mechanism. This test could detect as low as 20 copies of SARS-CoV-2 in one reaction. Additionally, the detection could be finished in 45 min. The test only needs a heating block and an ultraviolet lamp, which shows the potential for field detection.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序