Phenotypic and genotypic characterization of carbapenem encoding genes among carbapenem-resistant Gram-negative bacteria isolated from North Casablanca, Morocco

Carbapenem resistance genes in Gram-negative bacteria (CR-GNB) are a major cause of critical infections and are considered an urgent public health concern. The present study aimed to describe the prevalence of CR-GNB and the dissemination of extended-spectrum beta-lactamase (ESBL) and carbapenemase genes in clinical isolates from Casablanca, Morocco. Firstly, the strains were collected and identified using phenotypic and biochemical methods, then the antibiotic susceptibility was evaluated by the disc diffusion assay to screen isolates resistant to carbapenems. Secondly, three traditional methods, the carbapenem inactivation method, the modified Hodge, and the in-house carba-NP, were performed to predict the carbapenemase production by the included strains. Finally, conventional PCR was utilized to validate and detect the carbapenemase- and ESBL-related genes. Concerning the results, out of the identified 122 strains, 48 were CR isolates, including 30 Klebsiella pneumoniae, 13 Escherichia coli, and 5 Pseudomonas aeruginosa. Furthermore, these strains presented a high level of resistance. Moreover, the prediction of carbapenemase production by the phenotypic methods showed variable results. Also, the PCR analysis revealed a high occurrence of β-lactamase (ESBL and carbapenemase) genes in the included clinical strains, and most strains harbored multiple resistance genes. Our findings suggest that the three existing methods have some limitations, and a validation study is still necessary for the carbapenemase diagnostics.