Klebsiella Pneumoniae is globally responsible for hospital- and community-acquired infections. This study aimed to determine the prevalence of K. pneumoniae and investigate the antibiotic resistance profile among clinical specimens at Azadi Teaching Hospital in Kirkuk, Iraq, and detect the rpoB gene for molecular identification of K. pneumoniae in comparison with phenotypic and biochemical methods. In total, 250 clinical specimens were collected from patients in Azadi Teaching Hospital in Kirkuk, Iraq, between January 2018 and May 2018. The isolates were identified by morphologic and biochemical testing. Kirby-Bauer disk diffusion method was used in the antibiotics susceptibility test. Following that, 19 (7.6%) K. pneumoniae isolates were isolated from 250 clinical specimens (5 [5.61%] and 14 [8.69%] from males and females, respectively), and most of them (n=12; 11.76%) were isolated from the age group of 10-35 years old. The isolates were reported high resistance towards various types of antibiotics, especially penicillins and cephalosporins. In contrast, K. pneumoniae showed very low resistance to imipenem and amikacin (5.26% and 10.52%, respectively). The range of multidrug-resistant K. pneumoniae isolates in this study was estimated at 100%. In gene detection, all isolates in this study showed PCR product with 108 bp by K. pneumonia specific primer (rpoB). Developed antibiotic policies and regular surveillance of antibiotic susceptibility patterns may help to overcome the indiscriminate use of antibiotics that is a major cause of the emergence of drug resistance among pathogens.