Background: Emergence of carbapenemase producing microorganisms, specifically the New Delhi metallo-β-lactamase (NDM) and its different groups, in different areas all over the world, raised a global health concern. New Delhi metallo-beta-lactamase destructs carbapenems, that are considered one of the last lines of treatment for infections caused by resistant strains. Aim: Our study aimed for detection of the NDM gene among the carbapenem resistant Klebsiella spp. isolated from a variety of clinical samples and detection of the associated risk factors for acquiring such infection. Methods: Two hundred carbapenem-resistant Klebsiella isolates were enrolled and subjected to microbiological identification and antibiotic sensitivity testing. Isolates were gathered from 560 diverse clinical specimens collected from various medical departments in Ain Shams University Hospital. Klebsiella spp. isolates were exposed to Imipenem-EDTA combined disk method for the identification of metallo-β-lactamase (MBL) production, at that point real time polymerase chain reaction (RT-PCR) was done for the detection of the New Delhi metallo-β-lactamase (bla_NDM-1) gene among MBL phenotypic producing organisms. Results: Using Imipenem-EDTA, 82 Klebsiella isolates were detected to be MBL producers. The PCR showed that the blaNDM-1 gene is carried by 80 of the 82 MBL positive isolates (97.6%). There was no statistical significance regarding the risk factors and the gene acquisition. Conclusion: Real time-PCR used for detection of MBL is more sensitive than the phenotypic detection methods. There was no specific risk factor identified for acquiring of bla_NDM-1 gene in this study. Increasing in the  incidence of MBL prompt the need to evolve preventive measures to reduce their spread.