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The pathogenicity of Klebsiella is a major problem in most hospitals because of their resistance to multiple antibiotics and potential transfer of plasmids to other organisms. Isolation and enumeration of different bacterial species were made from 250 clinical samples collected from various sites of different patients in Chennai of India during 2013. The samples were subjected to different processes in order to isolate the causative agents of different diseases. The isolated organisms were Klebsiella, E. coli, Staphylococcus, and Salmonella, among these Klebsiella was the dominant one comprised 63% of total positive samples. Out of 75 clinical samples, 95 were positive and the rest (155) were sterile samples. Antibiotic sensitivity as well as determination of minimum inhibitory concentration (MIC) of antibiotic-Cefotaxime on Klebsiella species was done by tube dilution. The organisms resistant to all were categorized in 1st line of antibiotics, those arei ntermediate sensitive were categorized in 2nd line of antibiotics and finally the rem aining were kept for sensitive with 3rd line of antibiotics. It was found the antibiotics like Cefotaxime, Amykacin, Netilmycin, Cetpirane, Cifriaxone, Ampisulbactum, Azithromycin and Gentamycin were useful for most Klebsiella infection which arrested bacterial cell wall synthesis, in turn inhibited the bacterial growth.