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Emerg Infect Dis. Genomic diversity of oseltamivir-resistant influenza virus A (H1N1), Luxembourg, 2007?08

Giuseppe

Emeritus
Genomic Diversity of Oseltamivir-Resistant Influenza Virus A (H1N1), Luxembourg, 2007?08 (EID, excerpts, edited)

DOI: 10.3201/eid1509.090452

Suggested citation for this article: Gerloff NA, Kremer JR, Mossong J, Opp M, Muller CP. Genomic diversity of oseltamivir-resistant influenza virus A (H1N1), Luxembourg, 2007?08 [letter]. Emerg Infect Dis. 2009 Sep; [Epub ahead of print]

Genomic Diversity of Oseltamivir-Resistant Influenza Virus A (H1N1), Luxembourg, 2007?08

[Original Full Document: LINK. EDITED.]


To the Editor:

The prevalence of oseltamivir-resistant influenza viruses A (H1N1) (ORVs) increased dramatically worldwide during the winter of 2007?08 (1). Recent reports indicated that by early 2009 most influenza virus (H1N1) strains were resistant to oseltamivir (2). Resistant viruses were transmitted readily and were as viable and pathogenic as oseltamivir-sensitive viruses (OSVs) (3,4). The His275Tyr (N1 numbering) mutation in the neuraminidase (NA) genes of influenza virus A (H1N1) that confers resistance to oseltamivir has previously been associated with impaired virus replication, infectivity, and pathogenicity (5,6).

We investigated the genetic diversity in all 8 gene segments of representative ORVs and OSVs collected during December 2007?March 2008 by the National Influenza Sentinel Surveillance System in Luxembourg (www.lns.public.lu/statistiques/grippe).

Phylogenetic analyses were performed by using MEGA version 4.0 (7). Tree topology and posterior probabilities were calculated by using MrBayes version 3 (8). The sequences have been submitted to GenBank (accession nos. FM174406?60, FN401430?45, and FN401487?FN401518).

Among 140 viruses, 34 strains (24.3%) had the oseltamivir-resistant genotype (Tyr275) in the NA gene. Bayesian analyses of NA genes showed that ORVs formed a distinct cluster supported by high posterior probability (1.00) on the common node (Figure).

One resistant strain (LNS-365) was more closely related to OSVs (minimal Kimura distance 0.3%, 4 nt) than to ORVs (minimal Kimura distance 0.5%, 6 nt). In NA protein 33, ORVs showed the common Asp354Gly substitution in addition to the Tyr275 mutation. The resistant outlier LNS-365 encoded Asp354 like all other OSVs (n = 106). Similarly, only 4 other resistant strains from Europe from the same season shared Asp354 with all 2007?08 sensitive influenza virus (H1N1) strains (n = 251) available in public databases.

A total of 18?44 selected sequences from each of the other genes of ORVs and OSVs were generated to investigate which other genetic markers cosegregated with the resistant genotype.

Sequences derived from most of the other genes (PB1, polymerase A, hemagglutinin, nucleoprotein, matrix, nonstructural) of ORVs and OSVs were phylogenetically interspersed with no distinct clustering.

In contrast, matching the phylogeny of NA, PB2 sequences of genotypically resistant strains (n = 14) formed a distinct cluster supported by high posterior probabilities (1.00) and separate from all OSVs (n = 16) and the resistant outlier LNS-365 (Figure).

On the PB2 amino acid level, all OSVs and the resistant outlier LNS-365 shared Pro453, whereas all ORV encoded serine at the same position (Ser453). The outlier LNS-365 differed only by 2 aa from OSVs but by 4 aa from the closest resistant strain.

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