鲍曼不动杆菌
多重聚合酶链反应
生物
清脆的
底漆(化妆品)
多路复用
计算生物学
微生物学
基因
细菌
多重耐药
聚合酶链反应
抗生素
遗传学
化学
铜绿假单胞菌
有机化学
作者
Yufeng Wang,Yongcan Guo,Li Zhang,Yujun Yang,Shuangshuang Yang,Yang Liu,Huajian Chen,Chenggui Liu,Junjie Li,Guoming Xie
标识
DOI:10.1016/j.snb.2021.129600
摘要
Bacterial identification and phenotypic antibiotic susceptibility test (p-AST) limit timely anti-infection treatment of patients in clinic to a large extent. Here, we develop a rapid platform integrated multiplex PCR with CRISPR-Cas array to detect multidrug-resistant Acinetobacter baumannii (MDRAB). The platform relies on multiplex PCR amplification strategy which can simultaneously amplify the house-keeping gene and 4 β-lactamase genes of Acinetobacter baumannii (A. baumannii), accompanying with the indiscriminate single-stranded DNase activity of LbaCas12a to generate a single fluorescent signal for multiplex PCR products. The genotypic antibiotic susceptibility test (g-AST) of A. baumannii was completed within 2 h, with a detection limit down to 50 CFU/mL. In addition, we also proved the specificity to differentiate primer dimers. These findings jointly demonstrate the ultimate potential of the CRISPR-Cas12a method for multiple genes detection with high throughput and high specificity. Given the versatility and universality of CRISPR-Cas12a platform, it is expected that this research will further promote its application in the diagnosis and treatment of multidrug-resistant bacteria.
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