The increase in the prevalence of infections caused by certain bacteria, such asKlebsiella pneumonia(K. pneumoniae), is a global health concern. Bacterial production of an enzyme called extended-spectrum beta-lactamase (ESBL) can generate resistance to antimicrobial therapeutics. Therefore, between 2012 and 2013, we investigatedK. pneumoniaethat produce ESBLs with the prevalence of individual genes includingblaSHV,blaCTX-M,blaTEM, andblaOXAisolated from clinical samples. A total of 99 variable diagnostic samples including blood from hematological malignancies (n= 14) or other clinical sources including sputum, pus, urine, and wound (n= 85) were analyzed. All samples' bacterial type was confirmed and their susceptibility to antimicrobial agents was established. Polymerase chain reaction (PCR) amplification was carried out to ascertain presence of specific genes that includedblaSHV,blaCTX-M,blaTEM, andblaOXA.Plasmid DNA profiles were determined to assess significance between resistance to antimicrobial agents and plasmid number. It was found that among non-hematologic malignancy isolates, the highest rate of resistance was 87.9% to imipenem, with lowest rate being 2% to ampicillin. However, in hematologic malignancy isolates, the highest microbial resistance was 92.9% to ampicillin with the lowest rate of resistance at 28.6% to imipenem. Among collected isolates, 45% were ESBL-producers with 50% occurrence in hematologic malignancy individuals that were ESBL-producers. Within ESBL-producing isolates from hematologic malignancy individuals, blaSHVwas detected in 100%,blaCTX-Min 85.7%, andblaTEMandblaOXA-1at 57.1% and 27.1%, respectively. In addition,blaSHV,blaCTX-M, andblaOXAwere found in all non-hematological malignancy individuals withblaTEMdetected in 55.5% of samples. Our findings indicate that ESBLs expressingblaSHVandblaCTX-Mgenes are significantly prevalent inK. pneumoniaeisolates from hematologic malignancy individuals. Plasmid analysis indicated plasmids in isolates collected from hematological malignancy individuals. Furthermore, there was a correlation between resistance to antimicrobial agents and plasmids within two groups analyzed. This study indicates an increase in incidence ofK. pneumoniaeinfections displaying ESBL phenotypes in Jordan.Supplementary InformationThe online version contains supplementary material available at 10.1007/s44197-023-00096-2.