title: PCR deduction of invasive and colonizing pneumococcal serotypes from Venezuela: a critical appraisal.
abstract: BACKGROUND
  Serotype surveillance of Streptococcus pneumoniae is indispensable for evaluating the potential impact of pneumococcal conjugate vaccines. Serotyping by the standard Quellung reaction is technically demanding, time consuming, and expensive. A simple and economical strategy is multiplex PCR-based serotyping. We evaluated the cost effectiveness of a modified serial multiplex PCR (mPCR), resolving 24 serotypes in four PCR reactions and optimally targeting the most prevalent invasive and colonizing pneumococcal serotypes found in Venezuela.
  
  
  METHODS
  A total of 223 pneumococcal isolates, 140 invasive and 83 carriage isolates, previously serotyped by the Quellung reaction and representing the 18 most common serotypes/groups identified in Venezuela, were serotyped with the adapted mPCR.
  
  
  RESULTS
  The mPCR serotyped 76% of all the strains in the first two PCR reactions and 91% after four reactions, correctly identifying 17 serotypes/groups. An isolate could be serotyped with mPCR in less than 2 minutes versus 15 minutes for the Quellung reaction, considerably lowering labor costs. A restrictive weakness of mPCR was found for the detection of 19F strains. Most Venezuelan 19F strains were not typeable using the mPCR, and two 19F cps serotype variants were identified.
  
  
  CONCLUSIONS
  The mPCR assay is an accurate, rapid, and economical method for the identification of the vast majority of the serotypes from Venezuela and can be used in place of the standard Quellung reaction. An exception is the identification of serotype 19F. In this setting, most 19F strains were not detectable with mPCR, demonstrating a need of serology-based quality control for PCR-based serotyping.
authors.affiliation: Instituto de Biomedicina, Universidad Central de Venezuela, Caracas, Venezuela. teresitabego@gmail.com.
authors.forename: Teresita
authors.initials: T
authors.lastname: Bello Gonzalez
authors.forename: Ismar Alejandra
authors.initials: IA
authors.lastname: Rivera-Olivero
authors.forename: María Carolina
authors.initials: MC
authors.lastname: Sisco
authors.forename: Enza
authors.initials: E
authors.lastname: Spadola
authors.forename: Peter W
authors.initials: PW
authors.lastname: Hermans
authors.forename: Jacobus H
authors.initials: JH
authors.lastname: de Waard
mesh_terms: D002353:Carrier State / Q000382:microbiology*; D003362:Cost-Benefit Analysis; D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D011008:Pneumococcal Infections / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods; D065288:Serogroup; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D013997:Time Factors; D014687:Venezuela
chemical_list: D004269:DNA, Bacterial