Background: The aim of this study was to find out the prevalence of bacteria and their antimicrobial susceptibility pattern in hands of the laboratory workers. Laboratory associated infections are an occupational hazard for laboratory workers in the microbiology laboratory. The workers can expose to infection if they do not properly wash their hands before taking food. Materials: Swabs from 35 laboratory workers was taken before and after applying the different disinfectants. The swabs were directly inoculated onto blood agar, MacConkey agar and nutrient agar. Inoculated plates were incubated at 37ºC for 24 hours. The antibiotic sensitivity testing was done by Kirby Bauer disc diffusion method according to CLSI guidelines. Results: This study detects the major pathogenic bacteria in hands i.e. Staphylococcus aureus (40.58%), CoNS (21.74%), Klebsiella oxytoca and Pseudomonas aeruginosa (8.70%) were isolated. Conclusion: This study helps to minimize the infections by proper hand washing and also minimizing the spread of infection from one person to others.