Pneumococcal diseases caused by the bacterium Streptococcus pneumoniawere a worldwide public health problem.. According to the World Health Organization, community-acquired pneumonia was the main cause of pediatric death. This study was curriedatKing Saud University during 2008- 2010 in Saudi Arabia.Four hundred eighty eight isolates of Streptococcal pneumonia were kindly supplied by Al Kharj Military Hospital and these isolates represented a part of a large collection maintained at -70 ⁰C.  Their sources of isolation were infected patients of different ages. The viability of the isolates was tested on blood agar. Unfortunately only 266 isolates were viable. The viable isolates were serogrouped by using latex agglutination method and were found to fall into 20 serogroups that matched the same serogroups listed in the source culture (488isolates).Out of these 20 serogroups, 170 isolates were chosen for serotyping because they were common in all serogroups.Multiplex polymerase chain reaction (PCR) with specific primer was used for serotyping.Real time PCR were also used applying DNA belonging to 19F serotypes using the specific primer 19F produced five distinct beaks and illustrated variable amount of DNA concentration and similar melting point.  The process was repeated using with known serotypes 19A and specific primer 19A. The products were four peaks and showed variable concentration of DNA and similar melting point.All serotypes detected in this study included in the available vaccine PCV-7   (4, 6B, 9V, 14,18C, 19F, 23F) which is currently being used in Saudi Arabia. But in this study we found two new serotypes (19A, 6A) which do not include in this PCV-7 vaccine.