Assessment of real-time PCR for diagnosis of Mycoplasma pneumoniae pneumonia in pediatric patients.
We developed a real-time PCR to detect Mycoplasma pneumoniae with a primer set designed for the 16S rRNA gene. Clinical samples (n=937) were collected from children with community-acquired pneumonia between April 2002 and March 2004 at 12 Japanese medical institutions. Sensitivity of real-time PCR was calculated as 10 colony-forming units per reaction tube using a pMP01 plasmid carrying a 225-bp target DNA fragment of the 16S rRNA gene in M. pneumoniae M129, a standard strain. Results, obtained within 2 h, were compared with those of conventional culture and serologic methods. Of all cases tested, 151 (16.4%) and 129 (13.8%) were positive for M. pneumoniae by real-time PCR and by culture, respectively. Among the 151 cases, almost all of those tested serologically by passive agglutination showed a rise in M. pneumoniae antibody titre between acute and convalescent sera. We conclude that this real-time PCR can identify M. pneumoniae rapidly and fulfills the need for rapid identification, high sensitivity, and high specificity.
Laboratory of Infectious Agents Surveillance, Kitasato Institute for Life Sciences, Kitasato University, and National Hospital Organization Tokyo Medical Center, Japan.
Miyuki
M
Morozumi
Akira
A
Ito
Somay Y
SY
Murayama
Keiko
K
Hasegawa
Reiko
R
Kobayashi
Satoshi
S
Iwata
Naohisa
N
Kawamura
Haruo
H
Kuroki
Eiichi
E
Nakayama
Takeshi
T
Tajima
Kimiko
K
Ubukata
D000293:Adolescent; D002648:Child; D015169:Colony Count, Microbial / Q000379:methods; D017714:Community-Acquired Infections / Q000175:diagnosis / Q000382:microbiology; D006801:Humans; D009177:Mycoplasma pneumoniae / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D011019:Pneumonia, Mycoplasma / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods
D012336:RNA, Ribosomal, 16S