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Background: CTX-M betalactamases have shown a rapid spread in the recent years among Enterobacteriaceae and have become the most prevalent Extended Spectrum Beta-Lactamases (ESBLs) in many parts of the world. The introduction and dissemination of antibiotic-resistant genes limits options for treatment, increases mortality and morbidity in patients, and leads to longer hospitalization and expensive costs. We aimed to identify the beta-lactamases circulating encoded by the genes <i>bla</i>_CTX-M-15, <i>bla</i>_SHV-1 and <i>bla</i>_TEM-1 in <i>Escherichia coli (E. coli)</i> and <i>Klebsiella pneumoniae (K. pneumoniae)</i> strains. Furthermore, we established the associated resistance phenotypes among patients hospitalized in the Intensive Care Unit (ICU) from County Clinical Emergency Hospital of Craiova, Romania. Methods: A total of 46 non-duplicated bacterial strains (14 strains of <i>E. coli</i> and 32 strains of <i>K. pneumoniae</i>), which were resistant to ceftazidime (CAZ) and cefotaxime (CTX) by Kirby–Bauer disk diffusion method, were identified using the automated VITEK2 system. Detection of ESBL-encoding genes and other resistance genes was carried out by PCR. Results. <i>E. coli</i> strains were resistant to 3rd generation cephalosporins and moderately resistant to quinolones, whereas <i>K. pneumoniae</i> strains were resistant to penicillins, cephalosporins, and sulfamides, and moderately resistant to quinolones and carbapenems. Most <i>E. coli</i> strains harbored <i>bla</i>_CTX-M-15 gene (13/14 strains), a single strain had the <i>bla</i>_SHV-1 gene, but 11 strains harbored <i>bla</i>_TEM-1 gene. The <i>mcr-1</i> gene was not detected. We detected <i>tet</i>(A) gene in six strains and <i>tet</i>(B) in one strain. In <i>K. pneumoniae</i> strains we detected <i>bla</i>_CTX-M-15 in 23 strains, <i>bla</i>_SHV-₁ in all strains and <i>bla</i>_TEM-1 in 14 strains. The colistin resistance gene <i>mcr-1</i> was not detected. The tetracycline gene <i>tet</i>(A) was detected in 11 strains, but the gene <i>tet</i>(B) was not detected in any strains. Conclusions. The development in antibiotic resistance highlights the importance of establishing policies to reduce antibiotic use and improving the national resistance surveillance system in order to create local antibiotic therapy guidelines.