title: Comparison of sputum and nasopharyngeal swab specimens for molecular diagnosis of Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella pneumophila.
abstract: BACKGROUND
  Differentiation of atypical pathogens is important for community-acquired pneumonia (CAP). In this study, we compared sputum and nasopharyngeal swabs (NPS) for use in detection of Mycoplasma pneumoniae (MP), Chlamydophila pneumoniae (CP), and Legionella pneumophila (LP), using Seeplex PneumoBacter ACE Detection Assay (PneumoBacter; Seegene).
  
  
  METHODS
  Sputum and NPS specimens were collected from patients in 15 hospitals. DNA was extracted from sputum using QIAamp DNA Stool Mini Kit (Qiagen) and from NPS using easyMAG (bioMérieux). Both types of specimens were evaluated by multiplex PCR using PneumoBacter. To determine the diagnostic performance of this assay, sputum samples were also tested using BD ProbeTec ET Atypical Pneumonia Assay (APA; Becton Dickinson).
  
  
  RESULTS
  Among 217 sputum and NPS, 20 (9.2%), 2 (0.9%), and 0 sputum were positive for MP, LP, and CP, respectively, whereas 8 (3.7%) NPS were positive for MP. The sputum APA test yielded 186, 206, and 204 interpretable results for MP, LP, and CP, respectively. Of these, 21 (11.3%) were positive for MP, 2 (1.0%) were positive for LP, and 0 samples were positive for CP. Compared to APA, the sensitivity and specificity of the sputum assay for MP were 95.2% and 100.0%, respectively, whereas for the NPS assay, these were 38.1% and 93.9%. Sputum testing was more sensitive than NPS testing (P=0.002). For LP and CP diagnosis, PneumoBacter and APA tests agreed 100%.
  
  
  CONCLUSIONS
  Specimen type is crucial and sputum is preferred over NPS for simultaneous detection of MP, LP, and CP using multiplex PCR in CAP.
authors.affiliation: Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.
authors.forename: Min-Chul
authors.initials: MC
authors.lastname: Cho
authors.forename: Hyewon
authors.initials: H
authors.lastname: Kim
authors.forename: Dongheui
authors.initials: D
authors.lastname: An
authors.forename: Miyoung
authors.initials: M
authors.lastname: Lee
authors.forename: Shin-Ae
authors.initials: SA
authors.lastname: Noh
authors.forename: Mi-Na
authors.initials: MN
authors.lastname: Kim
authors.forename: Young Pil
authors.initials: YP
authors.lastname: Chong
authors.forename: Jun Hee
authors.initials: JH
authors.lastname: Woo
mesh_terms: D023521:Chlamydophila Infections / Q000175:diagnosis; D016993:Chlamydophila pneumoniae / Q000235:genetics* / Q000302:isolation & purification; D017714:Community-Acquired Infections / Q000175:diagnosis*; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D016952:Legionella pneumophila / Q000235:genetics* / Q000302:isolation & purification; D007877:Legionnaires' Disease / Q000175:diagnosis; D060885:Multiplex Polymerase Chain Reaction; D009177:Mycoplasma pneumoniae / Q000235:genetics* / Q000302:isolation & purification; D009305:Nasopharynx / Q000382:microbiology*; D011019:Pneumonia, Mycoplasma / Q000175:diagnosis; D011933:Reagent Kits, Diagnostic; D013183:Sputum / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic