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.
Morozumi
Miyuki
M
Laboratory of Infectious Agents Surveillance, Kitasato Institute for Life Sciences, Kitasato University, and National Hospital Organization Tokyo Medical Center, Japan.
Ito
Akira
A
Murayama
Somay Y
SY
Hasegawa
Keiko
K
Kobayashi
Reiko
R
Iwata
Satoshi
S
Kawamura
Naohisa
N
Kuroki
Haruo
H
Nakayama
Eiichi
E
Tajima
Takeshi
T
Ubukata
Kimiko
K
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