title: Target gene sequencing to characterize the penicillin G susceptibility of Neisseria meningitidis.
abstract: Clinical isolates of Neisseria meningitidis with reduced susceptibility to penicillin G (intermediate isolates, Pen(I)) harbor alterations in the penA gene encoding the penicillin binding protein 2 (PBP2). A 402-bp DNA fragment in the 3' half of penA was sequenced from a collection of 1,670 meningococcal clinical isolates from 22 countries that spanned 60 years. Phenotyping, genotyping, and the determination of MICs of penicillin G were also performed. A total of 139 different penA alleles were detected with 38 alleles that were highly related, clustered together in maximum-likelihood analysis and corresponded to the penicillin G-susceptible isolates. The remaining 101 penA alleles were highly diverse, corresponded to different genotypes or phenotypes, and accounted for 38% of isolates, but no clonal expansion was detected. Analysis of the altered alleles that were represented by at least five isolates showed high correlation with the Pen(I) phenotype. The deduced amino acid sequence of the corresponding PBP2 comprised five amino acid residues that were always altered. This correlation was not complete for rare alleles, suggesting that other mechanisms may also be involved in conferring reduced susceptibility to penicillin. Evidence of mosaic structures through events of interspecies recombination was also detected in altered alleles. A new website was created based on the data from this work (http://neisseria.org/nm/typing/penA). These data argue for the use of penA sequencing to identify isolates with reduced susceptibility to penicillin G and as a tool to improve typing of meningococcal isolates, as well as to analyze DNA exchange among Neisseria species.
authors.affiliation: Neisseria Unit, Institut Pasteur, 28 Rue du Dr Roux, Paris, France. mktaha@pasteur.fr
authors.forename: Muhamed-Kheir
authors.initials: MK
authors.lastname: Taha
authors.forename: Julio A
authors.initials: JA
authors.lastname: Vázquez
authors.forename: Eva
authors.initials: E
authors.lastname: Hong
authors.forename: Desiree E
authors.initials: DE
authors.lastname: Bennett
authors.forename: Sophie
authors.initials: S
authors.lastname: Bertrand
authors.forename: Suzana
authors.initials: S
authors.lastname: Bukovski
authors.forename: Mary T
authors.initials: MT
authors.lastname: Cafferkey
authors.forename: Françoise
authors.initials: F
authors.lastname: Carion
authors.forename: Jens Jørgen
authors.initials: JJ
authors.lastname: Christensen
authors.forename: Mathew
authors.initials: M
authors.lastname: Diggle
authors.forename: Giles
authors.initials: G
authors.lastname: Edwards
authors.forename: Rocío
authors.initials: R
authors.lastname: Enríquez
authors.forename: Cecilia
authors.initials: C
authors.lastname: Fazio
authors.forename: Matthias
authors.initials: M
authors.lastname: Frosch
authors.forename: Sigrid
authors.initials: S
authors.lastname: Heuberger
authors.forename: Steen
authors.initials: S
authors.lastname: Hoffmann
authors.forename: Keith A
authors.initials: KA
authors.lastname: Jolley
authors.forename: Marcin
authors.initials: M
authors.lastname: Kadlubowski
authors.forename: Amel
authors.initials: A
authors.lastname: Kechrid
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Kesanopoulos
authors.forename: Paula
authors.initials: P
authors.lastname: Kriz
authors.forename: Lotte
authors.initials: L
authors.lastname: Lambertsen
authors.forename: Ileanna
authors.initials: I
authors.lastname: Levenet
authors.forename: Martin
authors.initials: M
authors.lastname: Musilek
authors.forename: Metka
authors.initials: M
authors.lastname: Paragi
authors.forename: Aouatef
authors.initials: A
authors.lastname: Saguer
authors.forename: Anna
authors.initials: A
authors.lastname: Skoczynska
authors.forename: Paola
authors.initials: P
authors.lastname: Stefanelli
authors.forename: Sara
authors.initials: S
authors.lastname: Thulin
authors.forename: Georgina
authors.initials: G
authors.lastname: Tzanakaki
authors.forename: Magnus
authors.initials: M
authors.lastname: Unemo
authors.forename: Ulrich
authors.initials: U
authors.lastname: Vogel
authors.forename: Maria Leticia
authors.initials: ML
authors.lastname: Zarantonelli
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D005798:Genes, Bacterial; D005838:Genotype; D014943:Global Health; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000145:classification / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D010400:Penicillin G / Q000494:pharmacology*; D010403:Penicillin Resistance; D046915:Penicillin-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA
chemical_list: D000900:Anti-Bacterial Agents; D046915:Penicillin-Binding Proteins; D010400:Penicillin G