Sensitive and specific methods for rapid laboratory diagnosis of Mycoplasma pneumoniae were not available until nucleic acid amplification methods were developed. The choice of sample type and method of sampling are crucial to optimal diagnostic efficacy. Three types of respiratory samples from 32 young military conscripts with pneumonia were collected during an outbreak of M. pneumoniae infection. Sputum, nasopharyngeal aspirate and throat swab specimens were tested by 16S rRNA gene-based PCR with liquid-phase probe hybridization, and the results were compared with serology. The PCR result was positive for 22 (69 %) of the sputa, 16 (50 %) of the aspirates and 12 (37.5 %) of the swabs. Serology with increasing or high titres supported the positive findings in all instances. Sputum, when available, is clearly the best sample type for young adults with pneumonia.