Molecular typing and characterization of TEM, SHV, CTX-M, and CMY-2 β-lactamases in Enterobacter cloacae strains isolated in patients and their hospital environment in the west of Algeria.

OBJECTIVES

Enterobacter cloacae is a major nosocomial bacterium causing severe infections. A multicenter retrospective cohort study was conducted to collect baseline information on the molecular characteristics of β-lactamase producing Enterobacter cloacae in the west of Algeria.

MATERIALS AND

METHODS

We report a series of 42 extended-spectrum β-lactamase (ESBL) producing non-repetitive Enterobacter cloacae strains, collected in 3 university hospital (Tlemcen, Oran, and Sidi Bel Abbes). Antibiotic susceptibility testing (antibiogram and MIC) and screening for ESBL were performed according to the French Society for Microbiology guidelines. PFGE typing was used to characterize the clonality of all the strains. β-lactamase genes (blaTEM, blaSHV, blaCTX-M, blaECB, and blaCMY-2) were amplified by PCR with specific primers. Plasmid isolation, electroporation, and conjugation experiments were carried out using standard methods.

RESULTS

Sequence analysis revealed that most strains produced CTX-M type ESBLs (CTX-M-15 and CTX-M-3), whereas only 5 produced SHV-type ESBLs (SHV-12). The blaTEM gene was identified in all strains of Enterobacter cloacae. Several epidemic clones were determined. One strain was found to produce plasmid-mediated AmpC β-lactamase (CMY-2); this gene was transferred from E. cloacae by electroporation. Conjugation experiments showed that blaCTX-M, blaTEM, and blaSHV were carried by conjugative plasmids of high molecular weight (≥70kb).

CONCLUSION

The emergence of resistance genes is a public health problem.