Streptococcus pneumoniae is a prominent pathogen in children under age 5 and in elderly people, particularly in developing countries. A total of 100 S. pneumoniae isolates (clinical and normal flora) were obtained in Tehran, Iran. By using mPCR, 91 and 7 isolates were typed in the first and second reaction, respectively. Two isolates were identified as non-typeable. The most frequent serotypes were 23F (n=20, 20%), 3 (n=15, 15%), and 19F (n=15, 15%).