Stethoscopes are important vehicle for transmission of nosocomial infection from one patient to others. The study showed assessment of bacteria from diaphragm of stethoscopes of health care workers in a tertiary care hospital, Navi Mumbai. In our study swabs were collected from stethoscopes by rubbing sterile, moist swabs on the entire surface of diaphragms. In the second part of the study, the same stethoscope diaphragm was cleaned with alcohol rubs, allowed to act for 3-5 minutes and again second swab was collected from the same stethoscopes. The swabs were directly inoculated on blood agar, MacConkey agar plates. Standard methods were followed for isolation and identification of bacteria. All Staphylococcus strain was tested for MRSA. Antibiotic sensitivity was performed by Kirby-Bauer disc diffusion method as per CLSI guidelines to study antibiotic susceptibility pattern.Our study highlights the need to disinfect the stethoscopes diaphragms by simply applying the alcohol rubs to prevent any spread of bacteria from patient to patients.