期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 334, 期 -, 页码 -出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2021.129600
关键词
Acinetobacter baumannii; Multidrug; Resistant; Multiplex PCR; CRISPR-Cas12a; Sensor array
资金
- NSFC [81802115, 82002255]
- China Postdoctoral Science Foundation [2018M643425]
- Natural Science Foundation of Chongqing [cstc2019jcyjmsxmX0203]
- Chongqing Technology Innovation and Application Demonstration Project [cstc2018jscxmsybX0010]
- Top Talent Project of Chongqing Medical University [BJRC201926]
By using multiplex PCR and CRISPR-Cas method, we have developed a rapid platform for detecting multidrug-resistant Acinetobacter baumannii, with genotypic antibiotic susceptibility test completed within 2 hours, high specificity in differentiating primer dimers, and the potential for further application in the diagnosis and treatment of multidrug-resistant bacteria.
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 beta-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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