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Updated: Feb 1, 2026
Neisseria meningitidis Infection of Induced Pluripotent Stem-Cell Derived Brain Endothelial Cells
Published on: July 14, 2020
Development of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African
Kanny Diallo1,2, Mamadou D Coulibaly1, Lisa S Rebbetts2
1Centre pour le Développement des Vaccins (CVD), Bamako, Mali.
Abstract:
Improved methods for the detection and characterization of carried Neisseria meningitidis isolates are needed. We evaluated a multiplex PCR algorithm for the detection of a variety of carriage strains in the meningitis belt. To further improve the sensitivity and specificity of the existing PCR assays, primers for gel-based PCR assays (sodC, H, Z) and primers/probe for real-time quantitative PCR (qPCR) assays (porA, cnl, sodC, H, E, Z) were modified or created using Primer Express software. Optimized multiplex PCR assays were tested on 247 well-characterised carriage isolates from six countries of the African meningitis belt. The PCR algorithm developed enabled the detection of N. meningitidis species using gel-based and real-time multiplex PCR targeting porA, sodC, cnl and characterization of capsule genes through sequential multiplex PCR assays for genogroups (A, W, X, then B, C, Y and finally H, E and Z). Targeting both porA and sodC genes together allowed the detection of meningococci with a sensitivity of 96% and 89% and a specificity of 78% and 67%, for qPCR and gel-based PCR respectively. The sensitivity and specificity ranges for capsular genogrouping of N. meningitidis are 67% - 100% and 98%-100% respectively for gel-based PCR and 90%-100% and 99%-100% for qPCR. We developed a PCR algorithm that allows simple, rapid and systematic detection and characterisation of most major and minor N. meningitidis capsular groups, including uncommon capsular groups (H, E, Z).
Insights
A new multiplex PCR algorithm improves the detection and characterization of Neisseria meningitidis carriage strains. This method enhances identification of major and minor capsular groups, including uncommon ones, crucial for meningitis belt surveillance.
Area of Science:
- Microbiology
- Molecular Biology
- Epidemiology
Background:
- Neisseria meningitidis carriage is a significant public health concern, particularly in the African meningitis belt.
- Existing methods for detecting and characterizing carriage strains have limitations in sensitivity and specificity.
- Improved diagnostic tools are essential for effective surveillance and control of meningococcal disease.
Purpose of the Study:
- To develop and evaluate a multiplex PCR algorithm for enhanced detection and characterization of Neisseria meningitidis carriage strains.
- To improve the sensitivity and specificity of PCR assays for identifying N. meningitidis species and capsular genogroups.
- To provide a rapid and systematic method for analyzing meningococcal isolates from the African meningitis belt.
Main Methods:
- Modification and creation of primers/probes for gel-based and real-time quantitative PCR (qPCR) assays targeting specific N. meningitidis genes (sodC, H, Z, porA, cnl, E).
- Optimization of multiplex PCR assays and testing on 247 well-characterized carriage isolates from six African meningitis belt countries.
- Sequential multiplex PCR assays were used for genogroup characterization (A, W, X, B, C, Y, H, E, Z).
Main Results:
- The developed PCR algorithm successfully detected N. meningitidis species using both gel-based and real-time multiplex PCR.
- Targeting porA and sodC genes achieved high sensitivity (96% qPCR, 89% gel-based) and specificity (78% qPCR, 67% gel-based) for meningococcal detection.
- Capsular genogrouping demonstrated high sensitivity (67%-100%) and specificity (98%-100%) for gel-based PCR, and (90%-100%) and (99%-100%) for qPCR, respectively.
Conclusions:
- A novel, simple, rapid, and systematic PCR algorithm was developed for N. meningitidis detection and characterization.
- The algorithm effectively identifies most major and minor capsular groups, including uncommon ones (H, E, Z).
- This tool is valuable for enhanced surveillance and understanding of N. meningitidis carriage dynamics in the African meningitis belt.
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