The present study was done to see the microbial flora in the environment (air and surface) of Nepal Medical College Teaching Hospital and the staffs working in the hospital. Altogether 160 environmental (air n = 43, surface n = 117) samples were collected and studied from different wards. Similarly 150 samples (48 nasal swabs, 48 throat swabs and 54 hand samples) from the staffs were collected and studied following the standard microbiological protocols. Gram +ve cocci were the most predominant ones among the bacterial isolates from the environment followed by gram +ve bacilli and gram -ve bacilli. Among fungal isolates, yeast were the most common isolates while Aspergillus spp. were the most frequently occurring mold. Out of 150 samples collected for the study of carrier pattern, 32 out of 54 samples collected were found to have Staphylococcus aureus in their hands, 1 had Escherichia coli. Other isolates were Bacillus spp., Micrococci and coagulase negative staphylococci. Similarly 21 (43.8%) out of 48 nasal samples were found to have S. aureus while none of the staffs were found to have beta-hemolytic streptococci in their throat. In the study, 1.6% environmental isolates and 5.7% carrier isolates of S. aureus were found to be Methicillin resistant.