Background: Urinary tract infections (UTIs) are a significant public health concern, and Klebsiella pneumoniae is a common cause of these infections. The rise of antibiotic resistance among K. pneumoniae strains necessitates accurate and timely identification for effective treatment. Aim: This study aimed to isolate and identify K. pneumoniae from urine samples of patients with UTIs. It also sought to evaluate the effectiveness of different diagnostic methods, including phenotypic, biochemical, Vitek 2, and molecular techniques. Method: The study was conducted between August 2023 and November 2024 in Babylon province, Iraq, encompassing both public and private healthcare facilities. A total of 100 urine samples were collected from patients with UTIs, covering a diverse range of age groups and both sexes. The study employed a combination of diagnostic approaches. Phenotypic and biochemical tests were performed for initial identification. The Vitek 2 automated system provided rapid and accurate identification. Molecular identification was carried out using PCR amplification of the 16S rRNA gene. Results: K. pneumoniae was isolated from 12% of the urine samples. The highest prevalence was observed in the 10-30-year age group, and females had a higher incidence compared to males. All isolates were successfully identified using all diagnostic methods employed in the study. Conclusion: The study demonstrated the significant role of K. pneumoniae in causing UTIs in the studied population. The findings highlight the importance of utilizing a combination of diagnostic approaches for accurate and timely identification of this pathogen, which is crucial for effective treatment and infection control.