To determine the major molecular mechanisms of macrolide resistance among Streptococcus pneumoniae isolates in Turkey, we examined a total of 151 isolates collected from different regions of Turkey. Overall, 40 (26.4%) isolates were resistant to erythromycin. The most common mechanism (38/40) was target site modification due to erm (B) genes. Only two isolates hat-bored the mef(A)/(E) efflux gene. A clonal spread of resistant strains could not be demonstrated by BOX-polymerase chain reaction. The results from this study have shown that the erm (B) gene is predominant in Turkish S. pneumoniae isolates, as in isolates from the rest of the world, and a clonal dissemination is not responsible for this resistance profile.