Background: Hospital acquired infections (HAI) are a major problem in each hospital. HAI remains a significant hazard for hospitalized patients and health care workers are potential sources of these infections. Health care workers can transmit pathogens through their hands and contaminated medical devices such as stethoscopes, Blood pressure cuff etc. Hence we intended to determine the level of bacterial contamination of stethoscopes using in present hospital.Methods: Total of 50 stethoscopes were sampled before and after cleaning with 70% isopropyl alcohol by using sterile swab soaked in sterile saline. Samples were processed as per standard microbiological procedures.Results: Out of 50 stethoscopes, 30 (60%) stethoscopes were showing significant bacterial colonization, 7 (14%) showed insignificant colonization, and in 13 (26%) stethoscopes no growth was observed. The bacteria isolated were Methicillin Resistant Staphylococcus aurues (MRSA) (14%), Methicillin Sensitive Staphylococcus aureus (MSSA) (12%), Coagulase Negative Staphylococcus (CNS) (14%), Klebsiella species (20%).Conclusions: Our study concluded that stethoscopes used by health care workers were colonized by both pathogens and non-pathogenic microorganisms. Strict protocol of cleaning the stethoscopes in between the patients will eliminate the colonization and there by spread of infections.