Regulation of Carbapenemase Gene Conjugation in Escherichia coli Clinical Isolates

Background: The purpose of this study is to raise awareness of the hazards of carbapenemase epidemics and provide theoretical support for preventing the spread of carbapenemase-producing organisms. Methods: A total of 893 non-duplicate E. coil strains were recruited from three major local hospitals. The carbapenemase genotype of each imipenem-resistant strain was analyzed. Molecular typing and homology analysis of the main carbapenemase-producing strains reveal the transmission mode of resistance genes. Through the conjugation experiment, the potential spreading risk of carbapenemase genes was analyzed. Extended-spectrum beta-lactamase genes and replicon detection of the conjugant carrying plasmid were performed. The unannotated Escherichia coli bacterial small non-coding RNAs (sRNAs) interacting with sdiA were predicted through a bioinformatics tool. The sRNAs overexpression and knockout strains were constructed, and the effect of sRNA on conjugation was analyzed. Results: A total of 8 carbapenemase-producing strains were detected (0.90%, 8/893). The main carbapenemase genotype was blaKPC -2 (7 strains). Multilocus sequence typing indicated that 7 E. coli isolates belonged to ST-10, ST-101, ST-131, ST-405, ST-410, and ST-1193, ST-2562, respectively. Homologous cluster analysis revealed that the sequence types among the 7 E. coli were high diversity. The blaKPC -2 genes were successfully transferred from these isolates to EC600 by conjugation. All transconjugant cells exhibited significantly reduced susceptibility to the imipenem. IncFII was the most common conjugative plasmid type (85.7%, 6/7). Bioinformatics predicted the interaction between RydB and sdiA. Further experiments found that the interaction between RydB and sdiA improved the bacterial conjugation rate between MG1655 and EC600. The regulation effect of RydB on E. coli conjugation was not affected by the replicon type and/or harboring resistance coding genotype in conjugative plasmids. Conclusion: Our findings emphasized the epidemiological characteristics of carbapenemase-resistant E. coli. A functional phenotype of the new sRNA RydB was identified, and the regulation effect of RydB on E. coli conjugation was improved.