The development of a 16S rRNA gene based PCR for the identification of Streptococcus pneumoniae and comparison with four other species specific PCR assays

BMC Infect Dis. 2010 Apr 29:10:104. doi: 10.1186/1471-2334-10-104.

Abstract

Background: Streptococcus pneumoniae is one of the most frequently encountered pathogens in humans but its differentiation from closely related but less pathogenic streptococci remains a challenge.

Methods: This report describes a newly-developed PCR assay (Spne-PCR), amplifying a 217 bp product of the 16S rRNA gene of S. pneumoniae, and its performance compared to other genotypic and phenotypic tests.

Results: The new PCR assay designed in this study, proved to be specific at 57 degrees C for S. pneumoniae, not amplifying S. pseudopneumoniae or any other streptococcal strain or any strains from other upper airway pathogenic species. PCR assays (psaA, LytA, ply, spn9802-PCR) were previously described for the specific amplification of S. pneumoniae, but psaA-PCR was the only one found not to cross-react with S. pseudopneumoniae.

Conclusion: Spne-PCR, developed for this study, and psaA-PCR were the only two assays which did not mis-identify S. pseudopneumoniae as S. pneumoniae. Four other PCR assays and the AccuProbe assay were unable to distinguish between these species.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriological Techniques / methods*
  • Humans
  • Pneumococcal Infections / diagnosis*
  • Polymerase Chain Reaction / methods*
  • RNA, Bacterial / genetics*
  • RNA, Ribosomal, 16S / genetics*
  • Sensitivity and Specificity
  • Streptococcus pneumoniae / genetics*
  • Streptococcus pneumoniae / isolation & purification*

Substances

  • RNA, Bacterial
  • RNA, Ribosomal, 16S