Multiplex-PCR for diagnosis of bacterial meningitis

Braz J Microbiol. 2019 Apr;50(2):435-443. doi: 10.1007/s42770-019-00055-9. Epub 2019 Feb 22.

Abstract

Considering the great lethality and sequels caused by meningitis, rapid diagnosis and prompt treatment initiation have a great impact on patient outcome. Here, we developed a multiplex-PCR for simultaneous detection of the four most prevalent bacterial pathogens directly in CSF samples. The multiplex-PCR was designed to detect the following genes: fbsA (Streptococcus agalactiae), lytA (Streptococcus pneumoniae), crtA (Neisseria meningitidis), p6 (Haemophilus influenzae), and 16S rRNA (any bacterial agent). The multiplex-PCR showed a DNA detection limit of 1 pg/μL. Among 447 CSF samples tested, 40 were multiplex-PCR positive, in which 27 and 13 had positive and negative bacterial culture, respectively. Our multiplex-PCR is fast, reliable, and easily implementable into a laboratory routine for bacterial meningitis confirmation, especially for patients who previously started antimicrobial therapy. Our molecular approach can substantially improve clinical diagnosis and epidemiological measures of meningitis disease burden.

Keywords: Bacterial pathogens; Cerebrospinal fluid samples; Meningitis; Multiplex-PCR.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Cerebrospinal Fluid / microbiology*
  • Child
  • Child, Preschool
  • Female
  • Haemophilus influenzae / genetics*
  • Haemophilus influenzae / isolation & purification
  • Humans
  • Infant
  • Infant, Newborn
  • Male
  • Meningitis, Bacterial / diagnosis*
  • Meningitis, Bacterial / microbiology
  • Multiplex Polymerase Chain Reaction / methods
  • Neisseria meningitidis / genetics*
  • Neisseria meningitidis / isolation & purification
  • Streptococcus agalactiae / genetics*
  • Streptococcus agalactiae / isolation & purification
  • Streptococcus pneumoniae / genetics*
  • Streptococcus pneumoniae / isolation & purification