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A novel real-time PCR assay for the simultaneous detection of the four main causes of bacterial meningitis

K Diallo1, Tls Amoikon2, K F Missa3

  • 1Centre Suisse de Recherches Scientifiques en Côte d'Ivoire (CSRS), Abidjan, Côte d'Ivoire.

International Journal of Infectious Diseases : IJID : Official Publication of the International Society for Infectious Diseases
|January 19, 2026
PubMed
Abstract

Insights

A new real-time PCR assay accurately detects four key meningitis pathogens: Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, and Streptococcus agalactiae. This multiplex test improves meningitis diagnosis, especially in resource-limited settings.

Area of Science:

  • Microbiology
  • Molecular Diagnostics
  • Infectious Disease

Background:

  • Meningitis diagnosis relies on identifying key bacterial pathogens.
  • Current multiplex assays fail to detect all four WHO priority pathogens: Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, and Streptococcus agalactiae.
  • Accurate and rapid detection is crucial for effective meningitis management.

Purpose of the Study:

  • To develop and evaluate a novel real-time PCR assay for simultaneous detection of four WHO priority meningitis pathogens.
  • To assess the performance of the multiplex assay compared to monoplex assays.

Main Methods:

  • Developed a real-time PCR assay using specific primers and probes for N. meningitidis (sodC), H. influenzae (dmsA), S. pneumoniae (SP2020), and S. agalactiae (cfb).
  • Tested 44 DNA samples, including type cultures.
  • Evaluated assay performance using monoplex and multiplex formats, determining limits of detection, sensitivity, specificity, and predictive values.

Main Results:

  • The multiplex PCR assay demonstrated equivalent performance to monoplex assays.
  • Achieved 100% sensitivity for all targets.
  • Specificities ranged from 91.7% to 100%, with positive predictive values (PPVs) from 72.7% to 100% and negative predictive values (NPVs) of 100%.
  • Limits of detection ranged from 24 to 66 genome copies/µl.

Conclusions:

  • The developed multiplex PCR assay is effective for rapid and accurate detection of major meningitis-associated bacteria.
  • This assay has significant potential for improving meningitis diagnosis in low- and middle-income countries (LMICs).
  • Further field validation with clinical specimens is necessary prior to implementation.
Keywords:
EfficiencyMultiplex real-time PCRNPVPPVSensitivitySpecificity

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