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Target gene sequencing to characterize the penicillin G susceptibility of Neisseria meningitidis. 2007
Muhamed-Kheir Taha, and
Julio A Vázquez, and
Eva Hong, and
Desiree E Bennett, and
Sophie Bertrand, and
Suzana Bukovski, and
Mary T Cafferkey, and
Françoise Carion, and
Jens Jørgen Christensen, and
Mathew Diggle, and
Giles Edwards, and
Rocío Enríquez, and
Cecilia Fazio, and
Matthias Frosch, and
Sigrid Heuberger, and
Steen Hoffmann, and
Keith A Jolley, and
Marcin Kadlubowski, and
Amel Kechrid, and
Konstantinos Kesanopoulos, and
Paula Kriz, and
Lotte Lambertsen, and
Ileanna Levenet, and
Martin Musilek, and
Metka Paragi, and
Aouatef Saguer, and
Anna Skoczynska, and
Paola Stefanelli, and
Sara Thulin, and
Georgina Tzanakaki, and
Magnus Unemo, and
Ulrich Vogel, and
Maria Leticia Zarantonelli
Neisseria Unit, Institut Pasteur, 28 Rue du Dr Roux, Paris, France. mktaha@pasteur.fr
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.
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European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology,
