清脆的
布鲁氏菌
生物传感器
重组酶聚合酶扩增
计算生物学
布鲁氏菌病
生物
布鲁氏菌
质粒
细菌
聚合酶链反应
微生物学
病毒学
遗传学
DNA
基因
生物化学
作者
Jian-Hao Xu,Jianfeng Ma,Yanwei Li,Lin Woo Kang,Bin Yuan,Shiqing Li,Jie Chao,Lianhui Wang,Jinglin Wang,Shao Su,Yuan Yuan
标识
DOI:10.1016/j.snb.2022.131864
摘要
Brucellosis is a disease that leads to long-term damage in human and livestock and is caused by Brucella spp., which is an important and dangerous pathogen. Accordingly, the rapid and accurate detection of Brucella spp. is critical for decreasing the rate of infection. Since the discovery of the trans-cleavage ability of Cas effector proteins, the CRISPR/Cas system has shown great potential in the development of next-generation biosensors for biomolecule detection. Here, we coupled the trans-cleavage activity of a CRISPR/Cas12a system with recombinase polymerase amplification (RPA) and used two analytical methods to develop a general sensing platform for Brucella spp. detection. The resultant RPA-CRISPR/Cas12a based fluorescent biosensor (F-CRISPR) and electrochemical biosensor (E-CRISPR) can detect as few as 2 copies/reaction of positive reference plasmid. Importantly, the present biosensors developed can determine four main Brucella strains and distinguish them from other non-target bacteria. Finally, these dual- biosensors based on RPA-CRISPR/Cas12a enable a rapid and accurate detection of Brucella spp. in milk (food) samples and blood (clinical) samples that is comparable to the traditional RT-PCR method, suggesting that this RPA-CRISPR/Cas12a sensing platform is a powerful tool in the early diagnosis of brucellosis.
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