title: Detection of Streptococcus pneumoniae and Haemophilus influenzae type B by real-time PCR from dried blood spot samples among children with pneumonia: a useful approach for developing countries.
abstract: BACKGROUND
  Dried blood spot (DBS) is a reliable blood collection method for storing samples at room temperature and easily transporting them. We have previously validated a Real-Time PCR for detection of Streptococcus pneumoniae in DBS. The objective of this study was to apply this methodology for the diagnosis of S. pneumoniae and Haemophilus influenzae b (Hib) in DBS samples of children with pneumonia admitted to two hospitals in Mozambique and Morocco.
  
  
  METHODS
  Ply and wzg genes of S. pneumoniae and bexA gene of Hib, were used as targets of Real-Time PCR. 329 DBS samples of children hospitalized with clinical diagnosis of pneumonia were tested.
  
  
  RESULTS
  Real-Time PCR in DBS allowed for a significant increase in microbiological diagnosis of S. pneumoniae and Hib. When performing blood bacterial culture, only ten isolates of S. pneumoniae and none of Hib were detected (3·0% positivity rate, IC95% 1·4-5·5%). Real-Time PCR from DBS samples increased the detection yield by 4x fold, as 30 S. pneumoniae and 11 Hib cases were detected (12·4% positivity rate, IC95% 9·0-16·5%; P<0·001).
  
  
  CONCLUSIONS
  Real-Time PCR applied in DBS may be a valuable tool for improving diagnosis and surveillance of pneumonia caused by S. pneumoniae or Hib in developing countries.
authors.affiliation: University Hospital Sant Joan de Deu, Esplugues, Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Selva
authors.forename: Rachid
authors.initials: R
authors.lastname: Benmessaoud
authors.forename: Miguel
authors.initials: M
authors.lastname: Lanaspa
authors.forename: Imane
authors.initials: I
authors.lastname: Jroundi
authors.forename: Cinta
authors.initials: C
authors.lastname: Moraleda
authors.forename: Sozinho
authors.initials: S
authors.lastname: Acacio
authors.forename: Melania
authors.initials: M
authors.lastname: Iñigo
authors.forename: Alien
authors.initials: A
authors.lastname: Bastiani
authors.forename: Manuel
authors.initials: M
authors.lastname: Monsonis
authors.forename: Roman
authors.initials: R
authors.lastname: Pallares
authors.forename: Quique
authors.initials: Q
authors.lastname: Bassat
authors.forename: Carmen
authors.initials: C
authors.lastname: Muñoz-Almagro
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D001800:Blood Specimen Collection / Q000379:methods; D002648:Child; D002675:Child, Preschool; D004269:DNA, Bacterial / Q000097:blood / Q000235:genetics; D003906:Developing Countries; D005260:Female; D006192:Haemophilus Infections / Q000097:blood / Q000175:diagnosis / Q000382:microbiology*; D019741:Haemophilus influenzae type b / Q000235:genetics* / Q000302:isolation & purification; D006761:Hospitals; D006801:Humans; D007223:Infant; D008297:Male; D009018:Morocco; D009073:Mozambique; D011018:Pneumonia, Pneumococcal / Q000097:blood / Q000175:diagnosis / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013296:Streptococcus pneumoniae / Q000235:genetics* / Q000302:isolation & purification; D013301:Streptolysins / Q000235:genetics
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; C500439:CpsA protein, Streptococcus pneumoniae; D004269:DNA, Bacterial; D013301:Streptolysins; C032290:plY protein, Streptococcus pneumoniae; C068101:bexA protein, Haemophilus influenzae