Background With the advent of and rise in antibiotic resistance globally, especially in postoperative patients, studying the antibiogram and associated factors is the need of the hour. The present study was undertaken to document the microbiological profile in postoperative orthopedic patients with the infected implant in situ and to observe the antibiotic susceptibility patterns of isolated organisms in such infections. Methods This study was conducted in the Department of Microbiology of a tertiary care hospital for six months after obtaining institutional ethical approval. A total of 236 samples from patients with orthopedic implant infections were received during the study period, out of which 53 samples with positive culture isolate were further analyzed for microbiological profile including biofilm production. All observations and demographics were recorded and analyzed using SPSS software version 21.0 (IBM Corp., Armonk, NY, USA) and represented in the form of graphs, data, and tables. Results and conclusion The study showed a culture positivity rate of 53 (23%) out of 236 samples, in which gram-negative isolates 36 (68%) were more than gram-positive isolates 17 (32%). The most common isolate was 17 (32%) of which the majority were MRSA 13 (76%), followed by 9 (17%) and Klebsiella pneumoniae 9 (17%). Out of the 53 isolates, 20 were biofilm producers. Biofilm-producing isolates were more resistant to tested routine antibiotics compared to non-biofilm. This study could represent the initial interdisciplinary effort in an ongoing process to better understand and manage orthopedic implant infections at the hospital, specifically focusing on infections related to orthopedic devices.