放大器
重组酶聚合酶扩增
金黄色葡萄球菌
底漆(化妆品)
清脆的
聚合酶链反应
Cas9
重组酶
生物
微生物学
病毒学
计算生物学
化学
遗传学
细菌
基因
重组
有机化学
作者
Haoyu Wang,Qian Wu,Chao Yan,Jianguo Xu,Xinsheng Qin,Jie Wang,Wei Chen,Yao Li,Liwei Huang,Panzhu Qin
标识
DOI:10.1016/j.snb.2022.132293
摘要
Bloodstream infections have become a major global public health problem with high mortality. Rapid and robust pathogen diagnostics are necessary for improving patient care. Herein, we report an assay for the detection of Staphylococcus aureus (S. aureus) based on the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 and recombinase polymerase amplification (RPA) integrated lateral flow biosensor (LFB), namely CCR-LFB. Unlike existing LFBs based on dual-labeled amplicon bridging, CCR-LFB uses CRISPR/Cas9-mediated specific recognition of the amplicons induced by S. aureus DNA to form complexes that bind gold nanoparticles (GNPs) to the test (T) line. This rational design can completely overcome primer-dimer interference without reducing amplification efficiency, thereby improving sensitivity and robustness. Under optimal experimental conditions, S. aureus as low as 102 CFU/mL can be visually judged and a detection limit of 63 CFU/mL was achieved by ImageJ analysis. Additionally, this method has been successfully used to detect S. aureus in real blood samples without any cross-reaction. We hope that this method can serve as a model tool to meet the growing need for point-of-care diagnosis of bloodstream infections.
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