Fosfomycin (Phosphomycin) a new antimicrobial substance was first introduced in 1969. It is characterized as bactericidal agent with a wide spectrum of antimicrobial activity, both to Gram-negative and Gram-positive bacteria. It was used for many years as a highly effective antimicrobial drug especially for the treatment of urinary tract infections (UTIs). But currently the evolution and spread of various mechanisms of antimicrobial drug resistance among common family, Enterobacteriaceae is increasing. This results into narrowing of the available therapeutic options, which is of increasing global concern. The objective of this study was to evaluate the current pattern of fosfomycin resistance in Enterobacteriaceae using Disc Diffusion method. Total of 100 Enterobacteriaceae spp. were isolated from different specimen received in Pathology department of Tertiary Care Hospital. All the specimen were subjected to gram staining and culture. The samples were inoculated on Blood and MacConkey agar. Urine samples were cultured on CLED media. Enterobacteriaceae were identified based on their colony morphology, gram staining and biochemical tests (Triple sugar iron agar, Sulphur indole motility agar, Citrate agar) as per standard protocols. Antibiotic sensitivity testing was done by Kirby-Bauer disc diffusion method as per CLSI 2021 guidelines. Susceptibility patterns of fosfomycin against Enterobacteriaceae were observed and results were reported. Most of the gram negative Enterobacteriaceae were resistant to commonly used antibiotics. A total of 100 Enterobacteriaceae isolates were studied, including Escherichia coli (56%), Klebsiella oxytoca (16%), Klebsiella pneumoniae (10%), Serratia spp. (8%), Proteus spp. (5%) and others (5%). Antimicrobial susceptibility rates were highest for fosfomycin (92.8%) Of the 100 isolates, 69% Enterobacteriaceae isolates were fosfomycin sensitive and 31% were fosfomycin resistant.