Simultaneous single-tube PCR assay for the detection of Neisseria meningitidis, Haemophilus influenzae type b and Streptococcus pneumoniae

Clin Microbiol Infect. 2005 May;11(5):386-90. doi: 10.1111/j.1469-0691.2005.01109.x.

Abstract

Rapid, accurate and inexpensive diagnosis of bacterial meningitis is critical for patient management. This study describes the development and evaluation of a multiplex PCR assay for the detection of Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae type b, which globally account for 90% of cases of bacterial meningitis. The single-tube assay, based on the ctrA, ply and bex targets, respectively, enabled detection of 5-10 pg DNA. When the assay was tested with clinical samples (n = 425), its sensitivity for the three targets was 93.9%, 92.3% and 88%, respectively, while the overall specificity and positive predictive value of the assay was 100%. The negative predictive value was 99.1-99.5%. The methodology permits rapid and accurate detection of the three main pathogens that cause bacterial meningitis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • DNA, Bacterial / analysis
  • DNA-Binding Proteins / genetics
  • Diagnosis, Differential
  • Haemophilus influenzae type b / genetics
  • Haemophilus influenzae type b / isolation & purification*
  • Humans
  • Membrane Transport Proteins / genetics
  • Meningitis, Bacterial / diagnosis*
  • Meningitis, Haemophilus / diagnosis
  • Meningitis, Meningococcal / diagnosis
  • Meningitis, Pneumococcal / diagnosis
  • Neisseria meningitidis / genetics
  • Neisseria meningitidis / isolation & purification*
  • Pneumococcal Infections / diagnosis*
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Streptococcus pneumoniae / genetics
  • Streptococcus pneumoniae / isolation & purification*
  • Streptolysins / genetics
  • Transcription Factors / genetics

Substances

  • Bacterial Proteins
  • CtrA protein, Caulobacter
  • DNA, Bacterial
  • DNA-Binding Proteins
  • Membrane Transport Proteins
  • Streptolysins
  • Transcription Factors
  • plY protein, Streptococcus pneumoniae
  • bexB protein, Haemophilus influenzae