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Detection of Bacterial Meningitis Pathogens by PCR-Mass Spectrometry in Cerebrospinal Fluid

Piao Jing-Zi, He Zheng-Xin, Chen Wei-Jun

    Clinical Laboratory
    |June 27, 2018
    PubMed
    Abstract

    Insights

    A novel PCR-mass spectrometry (PCR-Mass) assay enables simultaneous detection of key bacterial meningitis and tuberculosis pathogens. This method accurately identifies bacteria in cerebrospinal fluid, even after early antibiotic treatment.

    Area of Science:

    • Molecular Diagnostics
    • Infectious Diseases
    • Microbiology

    Background:

    • Acute bacterial meningitis is a severe infectious disease with high morbidity and mortality.
    • Accurate and rapid detection of causative pathogens is crucial for outbreak prevention and infection control.
    • DNA-based detection methods are needed for timely diagnosis of meningitis-causing bacteria.

    Purpose of the Study:

    • To develop and evaluate a novel PCR-mass spectrometry (PCR-Mass) assay.
    • To simultaneously detect four major meningitis-causing pathogens: Neisseria meningitidis, Streptococcus pneumoniae, Haemophilus influenzae, and Mycobacterium tuberculosis.
    • To compare the performance of PCR-Mass with real-time PCR using clinical cerebrospinal fluid samples.

    Main Methods:

    • A multiplex PCR-mass spectrometry (PCR-Mass) assay was designed to target specific DNA sequences of four pathogens.
    • A total of 138 cerebrospinal fluid (CSF) samples were analyzed, including culture-positive, culture-negative, and control samples.
    • Results from PCR-Mass were compared against those obtained from real-time PCR detection.

    Main Results:

    • The PCR-Mass assay successfully discriminated between the four target pathogens without cross-reactivity.
    • The limit of detection was 10^2 copies/reaction for S. pneumoniae, H. influenzae, and N. meningitidis, and 10^3 for M. tuberculosis.
    • An overall agreement rate of 85.7% was observed between PCR-Mass and real-time PCR for culture-positive CSF samples. The assay also detected pathogens in culture-negative samples from patients receiving early antibiotic treatment.

    Conclusions:

    • PCR-Mass is a viable molecular method for detecting bacterial meningitis and tuberculosis.
    • The assay is particularly useful for diagnosing suspected cases where early antibiotic treatment may hinder culture-based methods.
    • This technology aids in the timely identification of pathogens, improving patient management and infection control strategies.

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