Polymerase chain reaction (PCR) with primers directed against the 16S-rRNA gene of Mycoplasma pneumoniae was used to diagnose M. pneumoniae infections, and the results were compared with those of culture and serology methods. Eighty (22%) of 363 throat swab samples from patients with acute respiratory complaints gave positive results by using the PCR method. Sixty-seven (18.5%) of the samples were positive in culture method. Of 35 samples which were unreliable culture results due to contamination with other bacteria, 13 gave positive results in the PCR method. Of the 97 cases obtained throat swabs and paired sera, 28 (28.9%) showed positive results by the PCR assay, and 29 (29.9%) by serology method (particle agglutination test). The positive rate was increased to 36% by using both the PCR and the serology methods. From these results it was concluded that the PCR method is useful for laboratory diagnosis of M. pneumoniae infections.