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

Kanny Diallo, Mamadou D Coulibaly, Lisa S Rebbetts, Odile B Harrison, Jay Lucidarme, Kadidja Gamougam, Yenenesh K Tekletsion, Akalifa Bugri, Aliou Toure, Bassira Issaka, Marietou Dieng, Caroline Trotter, Jean-Marc Collard, Samba O Sow, Xin Wang, Leonard W Mayer, Ray Borrow, Brian M Greenwood, Martin C J Maiden, Olivier Manigart and for the MenAfriCar Consortium

PLOS ONE, 2018, vol. 13, issue 12, 1-16

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).

Date: 2018
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Persistent link: https://EconPapers.repec.org/RePEc:plo:pone00:0206453

DOI: 10.1371/journal.pone.0206453

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