Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 17 5 2025 10 14 741 750 Muhammad Shafiq Department of Biosciences, Comsats University, Islamabad, Pakistan Amnah Khan Department of Biosciences, Comsats University, Islamabad, Pakistan Muhammad Hyder Department of Biosciences, Comsats University, Islamabad, Pakistan Umme Farwa Department of Pathology, Fazaia Medical College, Air University, Islamabad, Pakistan Farhan Kursheed Department of Microbiology, PMAS Arid Agriculture University, Rawalpindi, Pakistan Sana Khan Department of Pathology, Pakistan Institute of Medical Sciences (PIMS), Islamabad, Pakistan Ayesha Siddiqa Department of Pathology, Pakistan Institute of Medical Sciences (PIMS), Islamabad, Pakistan Iftikhar Ahmed National Culture Collection of Pakistan (NCCP), Land Resources Research Institute (LRRI), National Agriculture Research Centre, Park Road, Islamabad, Pakistan Muhammad Saeed Department of Biosciences, Comsats University, Islamabad, Pakistan Muhammad Fiaz Department of Pathology, Pakistan Institute of Medical Sciences (PIMS), Islamabad, Pakistan 2024 12 25 2025 07 18 Background and Objectives: Global health is seriously threatened by the rise of carbapenem-resistant Enterobacterales (CRE). The blaNDM gene, a key carbapenemase coding gene, causes global health concern due to its multidrug resistance and easy spread through mobile genetic elements. This study aimed to identify and genetically characterize the blaNDM genes from uropathogens, its antibiotic susceptibility, and its correlation with global sequences. Materials and Methods: Urine samples were processed following microbiological guidelines. Isolates were identified using API-20E. Antibiotic susceptibility was tested using disc diffusion method, and bacterial DNAs were extracted for blaNDM gene sequencing for phylogenetic analysis. Results: CREs were detected in 11.92% (n=51) of the 428 Enterobacterales. Among CRE isolates, 45% (n=23) were positive for blaNDM gene harbored by Klebsiella pneumoniae (57%), followed by Escherichia coli (26%). Uropathogenic CRE, harboring blaNDM, revealed susceptibility of 34.78%, 60.87%, and 65.22% to amikacin, nitrofurantoin, and fosfomycin respectively. The blaNDM-5 variant was most common (69.57%), followed by blaNDM-1 (26.09%) and blaNDM-7 (4.35%). Phylogenetic analysis revealed that blaNDM variants exhibit diverse relationships with Pakistani and worldwide sequences. Conclusion: The significant presence of blaNDM in uropathogens, along with extensive antibiotic resistance, underscores the urgent need for continuous monitoring and antibiotic stewardship programs to manage the growing threat of CRE infections. https://ijm.tums.ac.ir/index.php/ijm/article/view/5216 https://ijm.tums.ac.ir/index.php/ijm/article/download/5216/1820