title: Pleural fluid nucleic acid testing enhances pneumococcal surveillance in children.
abstract: OBJECTIVE
  National surveillance of invasive pneumococcal disease (IPD) includes serotyping Streptococcus pneumoniae (SP) isolates from sterile site cultures. PCR is more sensitive and can identify more SP serotypes (STs) in culture-negative samples. The aim of this study was to determine whether enhanced surveillance of childhood empyema, using PCR, provides additional serotype information compared with conventional surveillance.
  
  
  METHODS
  Pleural fluid (PF) from children with empyema were cultured and tested by PCR to identify SP, targeting the autolysin gene (lytA). Multiplex PCR-based reverse line blot assay was used to identify SP STs. Corresponding IPD surveillance and serotype data were obtained from the National Notifiable Diseases Surveillance System (NNDSS).
  
  
  RESULTS
  Eighty-nine children with empyema, aged ≤16 years, were recruited between April 2008 and March 2009, inclusive. SP was isolated from 5/84 (5.9%) PF cultures and by PCR in 43/79 (54.4%) PF samples. Serotypes were unidentifiable in 15 samples. The frequency of six serotypes (or serotype pairs) identified in 28 samples, including one with two serotypes, were: ST1, n = 4/29 (13.8%); ST3, n = 9/29 (31.0%); ST19A, n = 12/29 (41.4%); ST7F/7A, n = 1/29 (3.4%); ST9V/9A, n = 1/29 (3.4%); ST22F/22A, n = 2/29 (6.9%). Over the same period, 361 IPD patients, aged 16 years or less, were notified to NNDSS. Among 331 serotypeable NNDSS isolates (71.5% from blood), the frequencies of ST1 and 3 were significantly lower than in PF samples: ST1, n = 8/331 (2.4%; P < 0.05); ST3, n = 13/331 (3.9%; P < 0.0001).
  
  
  CONCLUSIONS
  The use of PCR to identify and serotype SP in culture-negative specimens provides additive information.
authors.affiliation: Department of Respiratory Medicine, Sydney Children's Hospital, Randwick, Australia.
authors.forename: Roxanne E
authors.initials: RE
authors.lastname: Strachan
authors.forename: Anita
authors.initials: A
authors.lastname: Cornelius
authors.forename: Gwendolyn L
authors.initials: GL
authors.lastname: Gilbert
authors.forename: Tanya
authors.initials: T
authors.lastname: Gulliver
authors.forename: Andrew
authors.initials: A
authors.lastname: Martin
authors.forename: Tim
authors.initials: T
authors.lastname: McDonald
authors.forename: Gillian
authors.initials: G
authors.lastname: Nixon
authors.forename: Rob
authors.initials: R
authors.lastname: Roseby
authors.forename: Sarath
authors.initials: S
authors.lastname: Ranganathan
authors.forename: Hiran
authors.initials: H
authors.lastname: Selvadurai
authors.forename: Greg
authors.initials: G
authors.lastname: Smith
authors.forename: Manuel
authors.initials: M
authors.lastname: Soto-Martinez
authors.forename: Sadasivam
authors.initials: S
authors.lastname: Suresh
authors.forename: Laurel
authors.initials: L
authors.lastname: Teoh
authors.forename: Kiran
authors.initials: K
authors.lastname: Thapa
authors.forename: Claire E
authors.initials: CE
authors.lastname: Wainwright
authors.forename: Adam
authors.initials: A
authors.lastname: Jaffé
mesh_terms: D000293:Adolescent; D001315:Australia / Q000453:epidemiology; D002648:Child; D002675:Child, Preschool; D016724:Empyema, Pleural / Q000276:immunology / Q000382:microbiology*; D005260:Female; D006801:Humans; D017589:Immunization Programs; D007223:Infant; D008297:Male; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics*; D011008:Pneumococcal Infections / Q000453:epidemiology / Q000276:immunology / Q000382:microbiology* / Q000517:prevention & control; D022242:Pneumococcal Vaccines; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D018571:Sentinel Surveillance; D013296:Streptococcus pneumoniae / Q000145:classification / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
chemical_list: D022242:Pneumococcal Vaccines; D009238:N-Acetylmuramoyl-L-alanine Amidase