Abstract: Objective Mycoplasma pneumoniae infection and provide evidence for the clinical diagnosis and treatment of M. pneumoniae infection. Methods A total of 346 clinical throat swabs were detected to conduct M.pneumoniae isolation and real-time PCR detection. Results Among the 346 specimens, the isolation rate of M.pneumoniae was 28.6% and the real-time PCR positive rate of M. pneumoniae was 32.1%. Among 205 community-acquired pneumonia (CAP) cases, the M. pneumoniae infection rate was 52.2%, but among 141 cases of upper respiratory tract infection (URTI), the detection rate of M. pneumoniae was only 4.2%, the difference was statistical significant. The difference in infection rate between children 14 years and adults 40 years had no statistical significance. Conclusion This study indicated that M. pneumoniae was a major pathogen to cause CAP, but was rare to be detected in URTI. Compared with the M. pneumoniae infection in children, the infection rate in adult was very high. More attention should be paid to it in clinical diagnosis.