Development of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African meningitis belt

PLoS One. 2018 Dec 5;13(12):e0206453. doi: 10.1371/journal.pone.0206453. eCollection 2018.

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).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms*
  • Female
  • Humans
  • Male
  • Mali / epidemiology
  • Meningitis, Meningococcal* / diagnosis
  • Meningitis, Meningococcal* / epidemiology
  • Meningitis, Meningococcal* / genetics
  • Multiplex Polymerase Chain Reaction / methods*
  • Neisseria meningitidis / genetics*
  • Neisseria meningitidis / isolation & purification
  • Porins / genetics*
  • Sensitivity and Specificity
  • Superoxide Dismutase / genetics*

Substances

  • Porins
  • porin protein, Neisseria
  • Superoxide Dismutase