title: Comprehensive detection of causative pathogens using real-time PCR to diagnose pediatric community-acquired pneumonia.
abstract: We have developed a real-time reverse transcription-PCR (RT-PCR) method to detect 13 respiratory viruses: influenza virus A and B; respiratory syncytial virus (RSV) subgroup A and B; parainfluenza virus (PIV) 1, 2, and 3; adenovirus; rhinovirus (RV); enterovirus; coronavirus (OC43); human metapneumovirus (hMPV); and human bocavirus (HBoV). The new method for detection of these viruses was applied simultaneously with real-time PCR for the detection of six bacterial pathogens in clinical samples from 1700 pediatric patients with community-acquired pneumonia (CAP). Of all the patients, 32.5% were suspected to have single bacterial infections; 1.9%, multiple bacterial infections; 15.2%, coinfections of bacteria and viruses; 25.8%, single viral infections; and 2.1%, multiple viral infections. In the remaining 22.6%, the etiology was unknown. The breakdown of suspected causative pathogens was as follows: 24.4% were Streptococcus pneumoniae, 14.8% were Mycoplasma pneumoniae, 11.3% were Haemophilus influenzae, and 1.4% were Chlamydophila pneumoniae. The breakdown of viruses was as follows: 14.5% were RV, 9.4% were RSV, 7.4% were hMPV, 7.2% were PIV, and 2.9% were HBoV. The new method will contribute to advances in the accuracy of diagnosis and should also result in the appropriate use of antimicrobials.
authors.affiliation: Kitasato Institute for Life Sciences, Laboratory of Molecular Epidemiology for Infectious Agents, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8641, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Hamano-Hasegawa
authors.forename: Miyuki
authors.initials: M
authors.lastname: Morozumi
authors.forename: Eiichi
authors.initials: E
authors.lastname: Nakayama
authors.forename: Naoko
authors.initials: N
authors.lastname: Chiba
authors.forename: Somay Y
authors.initials: SY
authors.lastname: Murayama
authors.forename: Reiko
authors.initials: R
authors.lastname: Takayanagi
authors.forename: Satoshi
authors.initials: S
authors.lastname: Iwata
authors.forename: Keisuke
authors.initials: K
authors.lastname: Sunakawa
authors.forename: Kimiko
authors.initials: K
authors.lastname: Ubukata
mesh_terms: D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D017714:Community-Acquired Infections / Q000175:diagnosis* / Q000382:microbiology / Q000821:virology; D004267:DNA Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006090:Gram-Negative Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006095:Gram-Positive Cocci / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D018410:Pneumonia, Bacterial / Q000175:diagnosis* / Q000821:virology; D011024:Pneumonia, Viral / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012328:RNA Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D055815:Young Adult