CRISPR-Hg: Rapid and visual detection of Hg2+based on PCR coupled with CRISPR/Cas12a

被引:4
作者
Kong, Fange [1 ]
Wang, Chunxia [1 ]
Peng, Shichao [1 ]
Chen, Zhengrui [1 ]
Huang, Yibing [2 ]
Zhang, Jicheng [4 ]
Wang, Jiasi [3 ]
Wang, Di [1 ]
机构
[1] Jilin Agr Univ, Chinese Minist Educ Edible & Med Fungi, Engn Res Ctr, Changchun City 130118, Jilin Prov, Peoples R China
[2] Jilin Univ, Sch Life Sci, Changchun 130012, Peoples R China
[3] Sun Yat Sen Univ, Sch Biomed Engn, Guangdong Prov Key Lab Sensor Technol & Biomed Ins, Shenzhen Campus, Shenzhen 518107, Guangdong Prov, Peoples R China
[4] Georgia Inst Technol, Sch Chem & Biomol Engn, Atlanta, GA 30332 USA
关键词
T-Hg2+-T triggered PCR; CRISPR/Cas12a; Trans-cleavage activity; Portable device;
D O I
10.1016/j.talanta.2024.126379
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Mercury (Hg) is a notorious toxic heavy metal, causing neurotoxicity and liver damage, posing grave threats to human health and environmental safety. There is an urgent imperative for developing novel Hg2+ detection methods. In this work, we developed a CRISPR-based method for Hg2+ detection named CRISPR-Hg. A CRISPR/ Cas12a system was employed and could be activated by the PCR product, generating fluorescence signals based on the trans-cleavage activity. CRISPR-Hg exhibited remarkable selectivity and specificity, achieving a detection limit of 10 pM and minimal interference with background signals. This approach has been successfully applied to detect Hg2+ in real samples, including water, soil, and mushroom. Ulteriorly, a portable device was devised to streamline the readout of fluorescence signals by a smartphone within 30 min. We offer an affordable, highly selective and visually interpretable method for Hg2+ detection, with the potential for broad application in Hg2+ monitoring for food safety and public health.
引用
收藏
页数:8
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