This study was designed to evaluate the serologic response to Mycoplasma pneumoniae infection. A total of 589 children < or =18 years (190 in the year 2004; 399 in 2005) and 2,073 adults > or = 18 years of age (980 in the year 2004; 1,093 in 2005) with respiratory symptoms underwent serological testing for M. pneumoniae infection. The tests included passive particle agglutination (PA) and enzyme-linked immunosorbent assay (ELISA). The seroprevalence rates of M. pneumoniae infection in the years 2004 and 2005 were 6.9 and 10.1%, respectively. The seropositivity rate was significantly higher in children (29.6% in 2005; 23.7% in 2004) than in adults (2.9% in 2005; 3.7% in 2004) (odds ratio, 8.138 in 2004; 13.923 in 2005; 95% confidence interval, 5.077-13.045 in 2004; 9.220-21.026 in 2005). Paired sera for the PA test were obtained from 32 of 399 children, and 22 of them demonstrated at least fourfold rises in antibody titer. ELISA had a sensitivity of 77.3% and a specificity of 40.0%; PA had a sensitivity of 9.5% and a specificity of 80%. The ELISA test was superior to the PA test in diagnosing acute M. pneumoniae infection in children. Both tests were significantly more sensitive when they were performed 1 week after the onset of infection.