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- Title
Rapid Detection of bla<sub>KPC</sub> in Carbapenem-Resistant Enterobacterales Based on CRISPR/Cas13a.
- Authors
Liang, Mingjun; Xiao, Bin; Chen, Lidan; Huang, Xiaoyan; Li, Jinchao; Kuang, Zhenzhan; Chen, Xinping; Huang, Xiuna; Sun, Zhaohui; Li, Linhai
- Abstract
Klebsiella pneumoniae carbapenemase (KPC) is a crucial enzyme that causes carbapenem resistance in Enterobacterales, and infections by these "superbugs" are extremely challenging to treat. Therefore, there is a pressing need for a rapid and accurate KPC detection test to control the prevalence of carbapenem-resistant Enterobacterales (CREs). In this study, we established a novel method for detection of blaKPC, the gene responsible for encoding KPC, based on a recombinase polymerase amplification (RPA) and a CRISPR/Cas13a reaction coupled to fluorophore activation (termed RPA-Cas13a assay). We carefully selected a pair of optimal amplification primers for blaKPC and achieved a lower limit of detection of approximately 2.5 copies/μL by repeatedly amplifying a recombinant plasmid containing blaKPC. The RPA-Cas13a assay demonstrated a sensitivity of 96.5% and specificity of 100% when tested on 57 blaKPC-positive CRE strains, which were confirmed by DNA sequencing. Moreover, in 311 sputum samples, the theoretical antibiotic resistance characteristics of blaKPC-positive strains obtained by the RPA-Cas13a assay were highly consistent with the results of antibiotic susceptibility test (Kappa = 0.978 > 0.81, P < 0.01). In conclusion, the RPA-Cas13a system is a simple and one-hour efficient technology for the detection of a potentially fatal antibiotic resistance gene.
- Publication
Current Microbiology, 2023, Vol 80, Issue 11, p1
- ISSN
0343-8651
- Publication type
Article
- DOI
10.1007/s00284-023-03457-z