AI Article Synopsis
- Klebsiella pneumoniae carbapenemase (KPC) is an enzyme responsible for making certain bacteria resistant to carbapenem antibiotics, making infections difficult to treat.
- This study introduced a new detection method for the KPC gene (bla) using a combination of recombinase polymerase amplification (RPA) and CRISPR/Cas13a technology, achieving high sensitivity and specificity in identifying the gene.
- The RPA-Cas13a assay proved efficient and rapid in detecting antibiotic resistance in clinical samples, showing strong alignment with traditional antibiotic susceptibility tests.
Article 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 bla, 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 bla and achieved a lower limit of detection of approximately 2.5 copies/μL by repeatedly amplifying a recombinant plasmid containing bla. The RPA-Cas13a assay demonstrated a sensitivity of 96.5% and specificity of 100% when tested on 57 bla-positive CRE strains, which were confirmed by DNA sequencing. Moreover, in 311 sputum samples, the theoretical antibiotic resistance characteristics of bla-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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10638124 | PMC |
| http://dx.doi.org/10.1007/s00284-023-03457-z | DOI Listing |
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