Simultaneous detection of Streptococcus pneumoniae and Haemophilus influenzae by nested PCR amplification from cerebrospinal fluid samples

FEMS Immunol Med Microbiol. 1997 Oct;19(2):151-7. doi: 10.1111/j.1574-695X.1997.tb01084.x.


Haemophilus influenzae and Streptococcus pneumoniae are often the cause of serious diseases such as meningitis. We designed a nested PCR assay to identify these pathogens from cerebrospinal fluid samples. The first-step PCR was able to detect eubacterial rRNA genes with a unified set of universal primers. In the second-step PCR, the identification primers, HI I and II and SP I and II, could detect H. influenzae and S. pneumoniae respectively through amplification of the rRNA spacer between the 16S and 23S rRNA genes. We suggest that the two-step PCR assay can be used as a novel method for the immediate and retrospective diagnosis of bacterial meningitis caused by H. influenzae and S. pneumoniae.

MeSH terms

  • Cerebrospinal Fluid / microbiology*
  • Child
  • Child, Preschool
  • Female
  • Haemophilus influenzae / isolation & purification*
  • Humans
  • Infant
  • Male
  • Meningitis, Bacterial / diagnosis
  • Polymerase Chain Reaction*
  • Streptococcus pneumoniae / isolation & purification*