Mycoplasma gallisepticum (MG) is major cause of chronic respiratory disease in chickens. M. synoviae (MS) most frequently occurs a subclinical upper respiratory infection but may result in airsacculitis and synovitis in chickens and turkeys. Both mycoplasmas induce economic losses by triggering chronic respiratory signs, airsacculitis and decreased egg production. For prevention of the infections, live attenuated andinactivated vaccines are commercially used for prevention of MG but not MS in Korea. Serum plate agglutination (SPA) and enzyme-linked immunosorbent assay (ELISA) have been commonly used for serological diagnosis for MG and MS. Recently, it is believed that MS spread in chickens is very seriously in Korea and respiratory infection with MS causes substantial loss in poultry farms. In this study, we investigated the serological prevalence of MG and MS in unvaccinated chickens between 2008 and 2009. The overall seroprevalence of MG was 24% of 2,094 for individual chickens and 24% of 189 farms. The overall seroprevalence of MS was 36% in 2,095 chickens and 39% in 198 farms. The results show that seropositive ratio of MS is higher than MG. The geographical prevalence of MG has been estimated in following sequence; Gangwon, Jeolla, Gyeonggi, Gyeongsang, and Chungcheong. The geographical prevalence of MS has been estimated as follows; Gangwon, Gyeonggi, Gyeongsang, Chungcheong, and Jeolla. Seasonal seroprevalencewas also examined, and it found that seroprevalence in spring, fall and winter was higher than that in summer in MG, but not in MS. No significant difference was shown in seroprevalence according to breed. Future study about pathogenicity of MS isolates would be needed and economical losses by MS outbreaks should be analyzed. Moreover, we compared sero-positivity obtained with SPA and ELISA. The kappa value of MG between SPA and ELISA was 0.8061 and the kappa value of MS between SPA and ELISA was 0.7649.