is an important cause of nosocomial infections in children. The study undertaken identified antibiotic resistance markers among biofilm-forming A total of 105 bacteremia-positive samples from hospitalized children were processed for identification of using species-specific gene. Phenotypic antibiotic resistance was checked through Kirby-Bauer disc diffusion method. 96-well microtiter plate assays and PCR were used for biofilm production and antibiotic-resistant genes, respectively. Among 105 clinical isolates, gene was detected in 34 (32.38%) isolates. The detected isolates exhibited biofilm formation ( = 34; 100%). Multidrug-resistant (MDR) pattern was observed among , while the frequency of MDR was higher in very strong biofilm-forming ( = 18; 52.9%, ≤ 0.002) as compared to weak biofilm-forming (6; 17.6%). All strains were resistant to cefoxitin, penicillin, and augmentin ( = 34; 100%). High resistance was observed against erythromycin ( = 29; 85.29%) and ciprofloxacin ( = 25; 73.5%). displayed complete susceptibility ( = 34; 100%) toward vancomycin, tetracycline, and linezolid. Among the isolates, the methicillin resistance gene (,  = 29; 85.2%, ≤ 0.000), the erythromycin resistance gene (,  = 19; 55.7%) and the beta-lactamase resistance gene (,  = 17; 50%) were detected. Detection of ( = 17; 94.4%), ( = 8; 44.4%) and ( = 11; 61.1%) significantly ( ≤ 0.0052) correlated with very strong biofilm-forming . Biofilm formation is significantly associated with antibiotic resistance. The study's result will help to understand the molecular mechanism of antimicrobial resistance in biofilm-forming among pediatric patients.