title: [Mycoplasma pneumoniae detection from throat swab by two-step polymerase chain reaction].
abstract: Two-step polymerase chain reaction (PCR) with primers designated against 16S rRNA gene of Mycoplasma pneumoniae for diagnosis of infection was evaluated in comparison with the conventional single-step PCR and culture methods. The two-step PCR method showed specific amplification of M. pneumoniae DNA and higher sensitivity (1.5 fg/assay) than the single-step PCR method. With the two-step PCR method, 76 of 322 throat swabs (23.6%) from patients with acute respiratory complaints gave positive results whereas 20.2% were positive in the culture method. Seven of 13 samples which were negative in the single-step PCR method but positive in either serological or the culture method showed positive results by the two-step PCR method. In addition, 5 samples which were weakly positive in the single-step PCR method showed distinctly positive results in the two-step PCR. These results indicate that the two-step PCR method is a useful tool for detection of M. pneumoniae in clinical specimens, although it requires a relatively sophisticated in technique.
authors.affiliation: Kanagawa Prefectural Public Health Laboratories, Department of Bacteriology and Pathology.
authors.forename: N
authors.initials: N
authors.lastname: Okazaki
authors.forename: S
authors.initials: S
authors.lastname: Yamai
authors.forename: Y
authors.initials: Y
authors.lastname: Sasaki
authors.forename: T
authors.initials: T
authors.lastname: Sasaki
mesh_terms: D006801:Humans; D009177:Mycoplasma pneumoniae / Q000302:isolation & purification*; D010614:Pharynx / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*