Background and Objectives: Despite improvements in modern diagnosis and therapies, hospital acquired infections remain a leading problem of global health systems. Healthcare Workers mobile phones is a reservoir for potential pathogens. Despite the high possibility of being contaminated, mobile phones are rarely clean and are often touched during or after examination of patients and handling of specimens without proper hand washing. The main objective of the present study was to isolate, identify different types of bacteria and their antibiotic sensitivity from mobile phones of healthcare Workers and non-healthcare Workers. Materials and Methods: Samples were collected aseptically by rolling over the exposed surfaces of the mobile phones inoculated on the agar plates and incubated aerobically. After incubation, plates were examined for growth. Bacteria were identified and antibiotic sensitivity was tested as per standard microbiological procedures. Results: In this study a total of 175 samples were examined, out of which 125 samples were from healthcare Workers (HCWs), 50 samples were from non-healthcare Workers (non-HCWs). Among the mobile phones of HCW’ s from ICUs, Acinetobacter baumannii (36. 84%) was the predominant organism isolated followed by methicillin resistant Staphylococcus aureus (MRSA) (21. 05%). Predominant organism isolated from HCW’ s in operation theater theater was MRSA (46. 66%). Out of 50 worker’ s non-HCWs mobile phones samples cultured, 23 (46. 00%) samples yielded growth of six different types of bacteria. Conclusion: Our study reveals that there is definite colonization of bacteria on mobile phones of the HCWs. It is not only capable of transferring message but also disease-producing microbes. In order to reduce incidence of nosocomial infections, there should be implementation of hand washing practices and regulations around the use of mobile telephones in hospital settings.