Background: Biofilm forming bacteria are the cause of many chronic and recurrent infections and are believed to be main factor for the development of non-healing wounds by inducing chronic inflammation. Therefore, early detection ofbiofilm producing bacterial infection along with their antimicrobial susceptibility pattern is needed for initiation of appropriate therapy. A prospective study was planned to detectbiofilm production in surgical site wound isolates and to compare antimicrobial susceptibility pattern of biofilm producing and biofilm non-producing isolates. Method: The present study was conducted for six months during which 170 pus isolates obtained from patients having surgical site wounds admitted in various wards in NSCB medical college, Jabalpur (M.P.) were studied. The organism was identified using standard microbiological procedures and AST was done by Kirby-Bauer disc diffusion method in accordance with CLSI guidelines 2023. Biofilm production was detected by Modified Tissue Culture Plate (MTCP) method. Result and Discussion: A total of 170 isolates were studied which included Staphylococcus aureus (n = 25), Coagulase negative Staphylococcus (n=21), Pseudomonas aeruginosa (n=44),Escherichia coli (n=20), Acinetobacterbaumannii (n=20), Klebsiellaspp. (n=15), Citrobacterspp. (n=13), Proteus spp. (n=08),Enterobacterspp. (n=04). Out of these, 63% isolates showed biofilm production. Also, multidrug resistance (MDR) was observed in 78% of the biofilm producing isolates and 40% of non- biofilm producing strains. The association of biofilm formation and multi drug resistance was found to be statistically significant. Conclusion: This study showed significantly high rate of biofilm formation in surgical site infection wound isolates. Also, it was observed that MDR strains were more commonly biofilm producers. Timely identification of these bacterial biofilms in surgical wounds will change the course of treatment thus helping in better patient management.