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Updated: Jun 25, 2026

Metagenomic Next-Generation Sequencing of Cerebrospinal Fluid for the Detection of Central Nervous System Pathogens
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Metagenomic Next-Generation Sequencing of Cerebrospinal Fluid for the Detection of Central Nervous System Pathogens

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Eight-plex PCR and liquid-array detection of bacterial and viral pathogens in cerebrospinal fluid from patients with

Mette Kusk Bøving1, Lisbeth Nørum Pedersen, Jens Kjølseth Møller

  • 1Department of Clinical Microbiology, Aarhus University Hospital, Skejby, Arhus, Denmark.

Journal of Clinical Microbiology
|February 6, 2009
PubMed

Insights

A new multiplex PCR assay accurately detects nine key meningitis pathogens in cerebrospinal fluid (CSF). This rapid test enhances early diagnosis and treatment for bacterial and viral meningitis.

Area of Science:

  • Clinical microbiology
  • Molecular diagnostics
  • Infectious disease epidemiology

Background:

  • Meningitis diagnosis relies on traditional methods like microscopy and culture, which can be slow.
  • Rapid and accurate identification of causative pathogens is crucial for effective meningitis treatment.
  • A need exists for a comprehensive assay to detect multiple common meningitis-causing microorganisms simultaneously.

Purpose of the Study:

  • To develop and validate a novel multiplex PCR assay for detecting nine significant bacterial and viral meningitis pathogens.
  • To evaluate the assay's performance against established diagnostic standards.
  • To assess the utility of this assay in a routine diagnostic setting for suspected meningitis cases.

Main Methods:

  • Development of a multiplex PCR assay targeting nine key pathogens: Neisseria meningitidis, Streptococcus pneumoniae, Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Streptococcus agalactiae, herpes simplex virus types 1 and 2, and varicella-zoster virus.
  • Detection of PCR products using the Luminex 100 suspension array system.
  • Validation using 1,187 cerebrospinal fluid (CSF) samples, comparing results to a gold standard of routine laboratory tests.

Main Results:

  • The multiplex PCR assay demonstrated excellent sensitivity and specificity for Streptococcus pneumoniae (95% and 99.1%) and Neisseria meningitidis (100% and 99.7%).
  • Out of 1,187 samples analyzed, 55 tested positive by the developed PCR assay.
  • The assay provides results within one workday, significantly faster than traditional methods.

Conclusions:

  • The novel multiplex PCR assay is a rapid, sensitive, and specific tool for identifying nine important meningitis pathogens in CSF samples.
  • This molecular method serves as a valuable supplement to microscopy and culture, aiding in timely and appropriate antimicrobial therapy selection.
  • The assay is suitable for initial screening and identification in patients with suspected meningitis, improving diagnostic workflow.

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