Ceftriaxone’s widespread utilisation in treating infections brought on by bacteria with multidrug resistance (MDR) has led to an increase in clinical resistance to the antibiotic in recent years. This research seeks to extract and characterise K. pneumoniae and A. baumannii bacteria, as well as examine the expression of Acr-B and Ade-B resistance genes for anti-Ceftriaxone resistance. Due to the collection of 170 human clinical samples from a diversity of sources. After being cultured, inspected microscopically, subjected to biochemical analysis, and put through the Vitec device, the bacteria revealed that 35 of the isolates were K. pneumoniae and 16 were A. baumannii. The MIC of ceftriaxone was then measured using the nutrient broth technique, and efflux pump gene expression was analysed using quantitative reverse transcription-PCR. Gene expression results for the Acr-B and Ade-B genes indicated that there was a change in the levels of expression of these genes after ceftrazone treatment of the bacterial isolates; the value of gene expression for the AcrB-Ab gene was 0.8 relative to the control gene, the amount of gene expression for the AcrB-gene Kp was (0.02), and the gene expression of the Ade-B-Kp gene was (10.33), i.e., an increase of (9. No gene expression was identified for the AdeB-Ab gene (the value was 0).