Amplification of P1 and 16S rRNA genes by nested PCR for detection of Mycoplasma pneumoniae in paediatric patients.

Mycoplasma (M.) pneumoniae is the most frequent atypical pathogen responsible for community-acquired respiratory infection in children and adults. The etiologic diagnosis of these infections still remains difficult. This is mainly due to the absence of characteristic clinical findings, and to the available detection methods (serology and culture) which are time consuming, insensitive and non-specific. To improve the detection of this infectious agent, nested polymerase chain reaction (PCR) analysis was developed. A total of 46 nasal aspirates, from children hospitalised with severe lower respiratory tract infection and in whom M. pneumoniae was suspected, were analysed for the presence of M. pneumoniae DNA by PCR. Routine microbiological investigations revealed no virus in these 46 samples. Using nested PCR, two targets were amplified: the sequences of 16S ribosomal (r) RNA gene (rDNA) and P1 adhesin gene. Evidence of M. pneumoniae infection was identified in four paediatric patients. The amplification of 16S rDNA was found to be more sensitive for the detection of M. pneumoniae. Our results suggest that amplification of the 16S rDNA by nested PCR and detection of the amplification products by visual inspection of the polyacrylamide gel should allow the rapid diagnosis of M. pneumoniae in respiratory tract infection in paediatric patients.