Comparative genomics of Neisseria meningitidis strains: new targets for molecular diagnostics.

Détails

Ressource 1Télécharger: BIB_79B332FB57D3.P001.pdf (505.27 [Ko])
Etat: Serval
Version: de l'auteur
ID Serval
serval:BIB_79B332FB57D3
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Comparative genomics of Neisseria meningitidis strains: new targets for molecular diagnostics.
Périodique
Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases
Auteur(s)
Diene S.M., Bertelli C., Pillonel T., Jacquier N., Croxatto A., Jaton K., Greub G.
ISSN
1469-0691 (Electronic)
ISSN-L
1198-743X
Statut éditorial
Publié
Date de publication
06/2016
Peer-reviewed
Oui
Volume
22
Numéro
6
Pages
568.e1-7
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Publication types: Comparative Study ; Evaluation Studies ; Journal Article

Résumé
In 2010, Jaton et al. (False-negative PCR result due to gene polymorphism: the example of Neisseria meningitidis. J Clin Microbiol 2010;48:4590-2) reported an isolate of Neisseria meningitidis serogroup B that was not detected by the ctrA quantitative real-time PCR (qRT-PCR) used in our diagnostic laboratory. Sequence analysis of ctrA revealed several single nucleotide polymorphisms responsible for the negative qRT-PCR. Therefore, we sequenced the genome of this isolate and performed comparative genomics to propose new gene targets for the specific detection of N. meningitidis from clinical specimens. We identified 11 genes as specific to N. meningitidis genomes and common to at least 177 (97%) of the 183 genomes available. Among them, three genes (metA, tauE and shlA) were selected to develop new qRT-PCRs for the detection of N. meningitidis DNA. The three qRT-PCRs were highly sensitive and specific, and they exhibited a good reproducibility when tested on plasmidic positive controls and genomic DNA extracted from strains of N. meningitidis and other relevant bacterial species. The clinical sensitivity and specificity of metA and tauE qRT-PCRs were both 100% based on a testing of cerebrospinal fluid samples positive for N. meningitidis or other clinically relevant bacteria. Despite a 100% specificity, the sensitivity of the shlA qRT-PCR was only 70%. We thus recommend using the metA and/or tauE qRT-PCRs developed here. To prevent PCR failure in the presence of new polymorphic strains, the detection of dual targets by duplex qRT-PCR would be more accurate and suitable for the diagnosis of N. meningitidis from clinical specimens.

Mots-clé
Child, Preschool, DNA, Bacterial/chemistry, DNA, Bacterial/genetics, Genes, Bacterial, Genome, Bacterial, Humans, Meningococcal Infections/diagnosis, Meningococcal Infections/microbiology, Molecular Diagnostic Techniques/methods, Neisseria meningitidis/classification, Neisseria meningitidis/genetics, Neisseria meningitidis/isolation & purification, Real-Time Polymerase Chain Reaction/methods, Sensitivity and Specificity, Sequence Analysis, DNA
Pubmed
Open Access
Oui
Création de la notice
03/05/2016 17:53
Dernière modification de la notice
08/05/2019 20:43
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